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The antioxidant activity of silkworm powder treated by proteolytic enzyme was investigated. Total protein content of silkworm power was assayed using BCA, Bradford assays and SDS-polyacrylamide gel electrophoresis (PAGE) with alkaline protease treatment conditions including temperature and pH. The optimum condition of alkaline protease treatment for silkworm powder was found to be 60℃ and pH 7. The alkaline protease treatment resulted in increased contents of free amino acids, total polyphenol and total flavonoid compared to control group. The silkworm hydrolysates showed excellent antioxidant activities in various in vitro models such as 2,2 diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, 2,2 - azino-bis(3-ethylbenzthiazoline-6)-sulfonic acid (ABTS) radical scavenging activity. These results provide useful information for using silkworm powder as an ingredient in functional foods and for exploiting alkaline protease treatment to improve the extractability and bioactivity of a raw material.

Hydrogels are crosslinked hydrophilic matrices for a variety of biomedical applications. Silk fibroin (SF), one of typical natural biomaterials, has been explored as base material for hydrogel. Photocrosslinked SF hydrogel containing poly(ethylene glycol) (PEG) was formulated through visible light initiated thiol-acrylate photopolymerization. The morphological, structural and thermal properties of SF . PEG hydrogel was investigated through scanning electron microscopy, X-ray diffractometry, thermogravimetry, and differential scanning calorimetry. The morphology of SF hydrogel showed dot and uneven surface with network cross-section. X-ray diffraction curves showed that the specific diffraction peaks of PEG were not changed by the intensity of the peaks were affected by sonication. Thermo-degradation behavior of SF . PEG hydrogel sonicated was significantly affected and became complex pattern compared to unsonicated ones. However, the melting endothermic temperature of SF-PEG hydrogel was not changed but the crystalline enthalpy was decreased by gelation and sonication.

There is no specific data on Entomogenous fungus (DongChungHaCho) in accordance with the silkworm varieties, despite of very high value industrial use as functional materials at domestic and abroad. In this study, we investigated culture condition and characteristics of Paecilomyces tenuipes in 3 silkworm cultivars and 2 specific silkworm breeds. Infection rate of P. tenuipes for silkworm varieties was the highest Kumokjam, followed by Baegokjam, Daeseongjam, Golden silk, Yeonnokjam in that order. The optimum culture conditions were as follows: dark condition, , and about in planting density per pupa. Also, we have developed annual production technologies of P. tenuipes. First, in order that the moisture content of the infection pupa became 3% or less in the natural condition. Second, the dried pupa must be vacuum-packed and was kept under . Finally, by restoring the moisture content of the dried pupa and culture in optimal condition, the technique for annual production of the P. tenuipes was achieved. Therefore, we expect that the use of Kumokjam and annual production technique will contribute significantly to farm income.

It is known that silk protein supports effectively proliferation of cell such as insect cell and hybridoma cell. Althoughthere are many varieties of Bombyx mori silkworm, the effect of silkworm varieties on cell proliferation has not beenconsidered in detail. We studied that characteristics of silk cocoon obtained from Baegokjam, Kumokjam, Daeseongjamsilkworm varieties and whether silk protein affected cell proliferation or not. Silk sericin was prepared under high tem-perature and high pressure condition. Silk fibroin was prepared using CaCl 2 :H 2 O : EtOH with different dissolutiontime. As a result, there are differences in silk cocoon from different silkworm varieties about cell proliferation. The pro-liferation was accelerated in the presence of Baegokjam silk sericin and Kumokjam silk fibroin with 5hr dissolutiontime. We expect that silk proteins could be a preferable culture medium supplement for stimulating the proliferation ofcell. Then, this results suggest silk as a new material for medium supplement replacing with fetal bovine serum.Key words : Silk fibroin, Silk sericin, Silkworm variety, Cell proliferation

Korean commercial Bombyx mori silkworm cocoon made by Kumokjam and Daesungjam was examined to characterize for application. Amino acid analysis showed that the main amino acid of Kumokjam and Daesungjam cocoon was glycine, alanine, serine, tyrosine, and aspartic acid. The content of three major amino acids was about 67% and the content of long chain amino acids was about 37%. Stress-strain curve showed that breaking stress of Kumokjam cocoon, 50 MPa, was higher than that of Daesungjam cocoon, 48 MPa. Breaking strain of Kumokjam cocoon, 37%, was also longer than that of Daesungjam cocoon, 30%. These results might be used as basic information for development of non-textile materials using Bombyx mori silkworm cocoon.

Enzymatic modification of proteins is often used to increase the biological activity of materials. Silkworm powder has been investigated as a functional food resource, but no study has been performed on its modification by commercial food enzyme. Therefore, this study aimed to determine the feasibility of such modification of silkworm powder by alkaline protease. The activity of the enzyme was confirmed using an azocasein assay. Subsequently the silkworm powder was hydrolyzed by enzymatic treatment. UV visible spectrometry showed that the supernatant of silkworm powder subjected to enzymatic treatment had a stronger absorption band than the untreated powder. SDS-PAGE electrophoresis showed that the molecular weight of silkworm powder decreased on enzymatic treatment. Thus the results indicate that commercial enzymes might be used to modify the characteristics of silkworm powder.

【Recently, many researchers have studied silk sericin because of its high water retention, good wound healing, good cyto-compatibility, and blood-glucose- and cholesterol-lowering effects. Although sericin film can be used in wound dressing and cosmetic packs, its poor mechanical properties have prevented its use in industrial fields. In the present study, sericin was obtained from different silkworm varieties, and the effect of silkworm variety on the structure and properties of sericin was examined. Except for a small difference in serine content, no significant difference in sericin was noted among the silkworm varieties. In addition, silkworm variety almost had no effect on solution viscosity, implying that it does not influence the molecular weight of sericin. Mechanical properties of sericin film were strongly affected by silkworm variety. Wonwon 126 showed the best mechanical properties, while N74 and Geumgwangju displayed the worst properties.】

This study aimed to investigate the effects of silk fibroin on bone metabolism in ovariectomized rats. A total of 30 Sprague-Dawley rats were randomized into sham-operated (SHAM), ovariectomized control (OVX), alendronate (OVX+ALEN, 10 mg/kg body weight/d), low silk fibroin (OVX+SF100, 100 mg/kg body weight/d), and high silk fibroin (OVX+SF300, 300 mg/kg body weight/d) groups. All the rats were fed by gavage for 12 wk. At the end of 12 wk, blood and urine were collected for analysis of bone turnover markers, and bone mineral density (BMD) was measured by micro-computed tomography. The results show that the OVX group (p < 0.05) displayed the highest mean body weight gain. Among the five groups, serum levels of bone alkaline phosphatase (ALP) and urine levels of deoxypyridinoline (DPD) were highest in the OVX group (p < 0.05). Bone ALP levels in the ALEN group were significantly lower than that of the silk-treated groups. On the other hand, DPD levels were not significantly different between the ALEN and silk-fibroin-treated groups ( p < 0.05). The trabecular BMD was significantly higher in the ALEN and silk-treated groups compared to the OVX group ( p < 0.05). In conclusion, this study showed that silk fibroin has similar effects as alendronate, which is used in osteoporosis medication. Therefore silk fibroin might be a new candidate for the prevention and treatment of osteoporosis in patients.

In this study, functionality of products of silkworm hemolymph fermented by Bacillus species was studied such as cellviability, antioxidant effect, and inhibitory effect on tyrosinase activities. A matured silkworm hemolymph was degradedby fermentation with Bacillus subtilis 10854 and Bacillus amyloliquefaciens M27. Especially, proteins of matured silk-worm hemolymph were degraded to 3,000 Da by fermentation with B. amyloliquefaciens M27. Cell viability for MTTassay was higher than PBS in hemolymph and fermented hemolymph. A DPPH free radical scavenging activity of fer-mented hemolymph was as higher as Vitamin C and dependent on sample concentrations. Thus, these results suggestthat degraded hemolymph fermented by B. amyloliquefaciens M27 may have antioxidant properties as a material forcosmetics.Key word : Silkworm, Hemolymph, Fermentation, Bacillus 서론 우리나라에 해외기술이 도입되고 화장품 연구개발이 본격적으로 이루어지기 시작한 1960년대 이후 국내 화장품시장 및 연구수준은 비약적인 발전을 이루었다. 2013년국내 화장품 시장 규모는 17조 6,000억으로 특히 국산화장품의 시장 점유율이 60%에 이르는 것으로 조사되었으며, 이는 국내에서 생산되는 원료로 다양한 과학기술을이용하여 생산한 화장품의 경쟁력이 높아지고 있다는 것을 의미한다. 최근에는 미백, 보습, 자외선 차단, 주름개선과 같은 화장품의 트랜드를 반영하기 위하여, 천연물의기능성을 활용하는 화장품 연구개발이 활발하게 이루어지고 있다. 흑마늘은 일반 마늘에 비해 생리활성물질의 증가로 새로운 기능성 소재로 각광받고 있으며 흑마늘의 티로시나아제(tyrosinase)와 엘라스타아제(elasetase) 활성 억제능과 자외선으로 인한 멜라닌 내 과산화물의 억제 효과에 대해 보고되어 있다(Jung et al. 2010, Lee et al. 2010).오미자씨 오일은 높은 안정성과 DPPH 라디칼 소거능 및콜라겐 합성능력이 뛰어나 기능성 화장품 소재로의 가능성이 보고된 바 있다(Yang 2012). 또한, 이러한 천연물을단순추출 및 사용하는 것 뿐만 아니라 미생물 발효에 의해 생성된 생리활성물질의 기능성을 평가하는 연구도 보고되고 있다. 균주로는 유산균(락토바실러스, Lactobacillus)과 효모(사카로미세스속, Saccharomyces), 고초균(바실러스속, Bacillus)을 주로 사용하고 있으며, Shim et al.(2012)에따르면 유산균으로 발효한 더덕 발효추출물이 pH 및 점도변화에 안정하고 경피 수분증발량을 억제하여 보습성을 높인다고 보고하였다. 마디풀 속과에 속하는 호장근(Polygonumcuspidata SIEB. et ZUCC.)을 효모균(Saccharomycescerevisiae)으로 발효시켜 SOD(Superoxide dismutase),catalase 항산화 효소활성을 측정한 연구에서 호장근 발효추출물은 농도 의존적으로 항산화 효소 활성이 증가하였다고 보고되었다(Lim et al. 2010). 바실러스속 세균은 자연계에 널리 존재하는 그람 양성의 간균으로 호기성 또는 통성 혐기성 조건에서 생육하는 것으로 알려져 있고주로 전분과 단백질 분해효소를 생성하는 것으로 연구되

We analyzed rutin content using mulberry genetic resources. They were grown under the same environment and con-ditions. Mulberry leaves were collected and then freeze-dried and powdered for rutin test. As a result, mean content ofmulberry strains was 0.52 ± 0.25%, and the coefficient of variation (CV) was 47.9%. The variation between the strainswas greatly severe. Among the tested strains, ‘Buyoungsang’ was showed the highest content of 1.37%, whereas‘Cheongeunsipmunja’ and ‘Mujeonsipmunja’ were showed the lowest content of 0.06% respectively. The content ofrutin of 16 mulberry varieties for silkworm rearing were compared. ‘Cheongolppong’ was showed the highest contentof 0.69%, whereas ‘Cheongilppong’ was showed the lowest content of 0.14 %. Finally we selected rutin high-containing9 strains. They are as follows. ‘Buyoungsang’, ‘Yeulbon’, 'Dangsang 5’, ‘A8’, ‘Aja’, ‘Seongsu 5’, ‘Sawonppong 12’,‘Hwanyoupjosaengnosang’ and ‘Hwansipjosaeng’ which are more than twice of the overall average content. Key words : Mulberry leaf, Rutin, Genetic resource

Propionibacterium acnes (P. acnes) is a major contributing factor to the inflammatory component of acne. The many prescription medications for acne allow for a large number of potential combination treatments. However, several antibiotics, apart from their antibacterial effects, exert side‑effects, such as the suppression of host inflammatory responses. Purified bee venom (BV) is a natural toxin produced by honeybees (Apis mellifera L.). BV has been widely used as a traditional medicine for various diseases. In the present study, to investigate the therapeutic effects of BV against P. acnes-induced inflammatory skin disease, P. acnes was intradermally injected into the ears of mice. After the injection, BV was applied to the skin surface of the right ear. Histological observation revealed that P. acnes induced a considerable increase in the number of infiltrated inflammatory cells. However, treatment with BV markedly reduced these reactions compared with the P. acnes-injected mice not treated with BV. Moreover, the expression levels of tumor necrosis factor (TNF)-α, and interleukin (IL)-1β were significantly reduced in the BV-treated mice compared with the untreated P. acnes-injected mice. In addition, treatment with BV significantly inhibited Toll-like receptor (TLR)2 and CD14 expression in P. acnes-injected tissue. The binding activity of nuclear factor-κB (NF-κB) and activator protein (AP)-1 was markedly suppressed following treatment with BV. The results from our study, using an animal model, indicate that BV exerts an inhibitory effect on inflammatory skin diseases. In conclusion, our data indicate that BV has potential for use as an anti-acne agent and may be useful in the pharmaceutical and cosmetics industries.

This research was performed in order to investigate the origin, standard compound, and structural and physical properties of honeybee venom which used as natural antibiotic ingredients to animal. We compared the nucleotide sequence of mitochondrial cytochrome c oxidase subunit 1 gene (COI) of honeybees were collected from Gangwon, Gyeonggi, Chungnam, Gyeongbuk, Gyeongnam province and Suwon. As major constituent of honeybee venom, melittin was assayed by liquid chromatography. X-ray, differential scanning calorimetry (DSC) and fourier transform infrared spectroscopy (FT-IR) were utilized to examine the structural and physical properties of honeybee venom. Based on the 627bp sequence of COI, Apis mellifera ligustica was determinated honeybees collected from all six regions. Melittin content varied from 50.7 to 68.6 and averaged 59.8%. According to XRD analysis, honeybee venom showed regular crystal structure peaks at 2θ=8.5° and 21.5°. DSC showed that the maximum degration temperature of powder was around 230°C. Through FT-IR analysis, we could identify cross-linking by the presence of peptide peak at 1,500∼1,600 cm −1 . In conclusion, the origin of honeybee venom was Apis mellifera ligustica and effective ingredient standards was melittin content varied from 50.7 to 68.6 as natural antibiotic ingredients.

In order to the feasibility of silk materials as protein delivery system, silk beads incorporated with bovine serum albumin (BSA) were prepared by dropping silk fibroin extract into dope solution composed of ethanol and dichloromethane. Structural and morphological characteristics of silk beads were examined using scanning electron microscopy (SEM), infrared spectrometry, and X-ray diffractometry. Swelling ratio of silk beads was also measured. Release behavior of prototypical protein, BSA, was studied by observing the electropheretic phenomenon and release profile. SEM showed that silk beads are spherical with porous interior structure. Infrared spectrometry and X-ray diffraction confirm that the silk beads have a -sheet conformation. The swelling capability of silk beads increased with the incorporation of the protein. The protein was released from the beads with slow release following an initial burst release. Therefore, silk beads show promise as materials for encasing protein drugs to be delivered to targets in the human body.

Purpose: This study evaluated powdered burn wound dressing materials fr om wild silkworm fibroin in an animal model.Methods: Fifteen rats were used in this experiment. Full-thickness 2×2 cm burn wounds were created on the back of rats under anesthesia. In the two experimental groups, the woun ds were treated with two different dressing materials made from silkworm fibroin. In the Control Group, natural healing wi thout any dressing material was set as control. The wound surface area was measured at five days, seven days and 14 days. Wound healing was evaluated by histologic analysis.Results: By gross observation, there were no infections or severe infla mmations through 14 days post-injury. The differences among groups were statistically significant at seven days and 1 4 days, postoperatively (P＜0.037 and 0.001, respectively). By post hoc test, the defect size was significantly smaller in experimental Group 1 compared with the Control Group and experimental Group 2 at seven days postoperatively (P=0.022 and 0.029, respectively). The difference between Group 1 and Group 2 was statistically significant at 14 days postoperat ively (P＜0.001). Group 1 and control also differed significantly (P=0.002). Group 1 showed a smaller residual scar than the Contro l Group and Group 2 at 14 days post-injury. Histologic analysis showed more re-epithelization in Groups 1 and 2 than i n the Control Groups.Conclusion: Burn wound healing was accelerated with silk fibroin spun by w ild silkworm Antheraea pernyi. There was no atypical inflammation with silk dressing materials. In concl usion, silk dressing materials can be used for treatment of burn wound.Key words:Full thickness burn wound, Wound dressing, Silk fibroin, Antheraea pernyi, Rats

This study was for the investigation of the stability of purified bee venom (PBV) during the treatment in the pH rangefrom pH2 to pH9 for 24 hours, respectively. Changes of components and physiological functionalities in PBV were byevaluated silver staining, and melittin contents were measured by liquid chromatography. The antimicrobial activityagainst bacteria by minimum inhibitory concentration (MIC) and effect of the cell regeneration were measured by 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl tetrazolium bromide (MTT) assay using human dermal fibroblast (HDF) cell.The main proteins such as melittin and phospholipase A 2 showed no characteristic changes. The antimicrobial activityand effect of cell regeneration showed no difference from pH2 to pH9. From this study, we suggest that componentsand physiological functionalities of PBV against treated pH were kept stability at from pH2 to pH9.Key words : Apis mellifera, Bee venom, Component, pH 서론 봉독은 꿀벌(Apis mellifera L.)의 독낭에 들어있는 자기방어 물질로 봉침은 수천년 전부터 관절염, 통풍 등(Kimet al. 2002, Kim et al. 2003) 치료에 한방과 민간요법으로 이용되어 왔다(Kim 1992). 봉독은 40여 가지의 성분으로이루어져 있으며, peptide 성분으로는 melittin, apamin, mastcell degranulating(MCD) peptide 등이 있으며, 효소 성분으로는 phospholipase A

Regenerated Antheraea pernyi silk sponge was prepared using calcium nitrate 4 hydrate melt and examined the conformational changes treated with aqueous ethanol solution. The conformation of silk sponges was changed from random coil structure to -sheet and -helix conformation with low ethanol concentration (50 ~ 70%). On the other hand, that of silk sponges with 80% ethanol treatment showed -sheet (), -helix (), and random coil () specific peaks. Wound healing effect in vitro was observed by cytoslective wound healing kit. Therefore, regenerated Antheraea pernyi silk sponges might be used as promising wound dressing materials.

Breast cancer is one of the most common cancers among women worldwide. Recently anticancer agents have been developed using natural substances. To evaluate the anticancer effect of hydrolysates of silk fibroin (HSF), we investigated the effect of HSF on cell viability and apoptosis of a breast cancer cell line, MCF-7, induced through the mitochondrial pathway. The result showed that HSF decreased cell viability in MCF-7 cells in a dose- and time-dependent manner, resulting in an increase in the sub-G1 phase cell population. HSF increased the level of the pro-apoptotic Bax protein and decreased the levels of the anti-apoptotic Bcl-2 protein. In addition, HSF induced apoptosis in MCF-7 cells through a mitochondria-dependent pathway by increasing levels of cytochtome c, and cleavage of PARP. Taken together, these findings suggest that HSF inhibits the proliferation of MCF-7 breast cancer cells through a mitochondria and caspase dependent apoptotic pathway.

In this study, we compared the efficiency of osteoblast differentiation media (ODM) contain ing three distinct reagent combinations in osteoblastic differentiation of human bone marrowderived mesenchymal stem cells (hBMSCs) in monolayer culture. In addition, we analyzed growth and differentiation of hBMSCs on silk scaffolds and examined the bone-forming activity of a nanofibrous silk scaffold in a tibia diaphysis defect model of a rat hind limb with intramed ullary nailing. Although all three ODM increased alkaline phosphatase activity to a comparable extent, the ODM containing bone morphogenetic protein-2 (BMP-2) was found to be signifi cantly less effective in promoting mineral deposition than the others. Growth of hBMSCs on sponge-form silk scaffolds was faster than on nanofibrous ones, while osteoblastic differentia tion was apparent in the cells grown on either type of scaffold. By contrast, bone formation was observed only at the edge of the nanofibrous scaffold implanted in the tibia diaphysis defect, suggesting that use of the silk scaffold alone is not sufficient for the reconstitution of the long bone defect. Since silk scaffolds can support cell growth and differentiation in vitro, loading MSCs on scaffolds might be necessary to improve the bone-forming activity of the scaffold in the long bone defect model.

In this study, matured silkworm hemolymph was studied for inhibitory effect on melanin formation, tyrosinase activities.A matured silkworm hemolymph mass purification method was developed and some of suppressing melanin formationeffects were investigated, too. As a result, the advanced purification method was higher in yield rate than existing oneby 53%. A free radical scavenging activity of hemolymph was higher than sericin on the market(IC 50 , 202.9± 33.1) andlower than vitamin C(0.57± 0.02) by 6.4± 0.3 ㎍/㎖. An inhibitory effect on melanin synthesis was higher inhemolymph compared to Morus bark and arbutin by 9.15% and 11.56%, respectively. An inhibitory effect on tyrosinaseactivity was higher in hemolymph than Morus bark and lower than arbutin. Effect of hemolymph 45㎍/㎖ on sup-pressing of tyrosinase was almost 90%. Thus, these results suggest that hemolymph of matured silkworm may have ben-eficial properties as a material for cosmetics.Key words : Matured silkworm hemolymph, Antioxidant, Melanin synthesis, Tyrosinase activity

In this study, bee venom (BV) isolated from honey bees Apis mellifera was assessed for its potential use as an antimicrobial agent against fish pathogenic bacteria. We used three bacterial isolates (Edwardsiella tarda, Vibrio ichthyoenteri, and Streptococcus iniae) that were originally obtained from the gastrointestinal tracts of moribund Olive Flounder Paralichthys olivaceus. Bee venom exhibited antibacterial activity against all three infectious fish pathogens. The minimum inhibitory concentration and minimum bactericidal concentration (mean ± SE) of BV were 17.6 ± 2.6 and 34.9 ± 3.4 μg/mL, respectively, against E. tarda; 1.76 ± 0.3 and 6.8 ± 2.6 μg/mL against V. ichthyoenteri; and 3.49 ± 0.9 and 11 ± 1.6 μg/mL against S. iniae. The postantibiotic effect of BV was 5 ± 0.6 h for E. tarda, 6 ± 0.3 h for V. ichthyoenteri, and 7 ± 0.6 h for S. iniae. In addition, the antimicrobial activity of BV was not pH dependent, as inhibition zones were identical at pH levels ranging from 2 to 11. These result...

Antheraea pernyi silkworm is one of typical wild silkworms, which spins a tawny color cocoon. The cocoon has been used as a resource for textile material due to strong chemical stability and good mechanical properties. In this study, to increase the solubility efficiency of A. pernyi silk fibroin, the composition of dissolution solvent were examined. Calcium chloride tertiary system, normally used for dissolution of Bombyx mori silk fibroin, does not act on A. pernyi silk fibroin. Calcium nitrate system dissolves A. pernyi silk fibroin, and calcium nitrate ethanol system do more easily than calcium nitrate system. Amino acid composition of A. pernyi silk fibroin obtained after dissolution is mainly composed of alanine, glycine, and serine. A. pernyi silk fibroin would be used for non-textile applications near future.

The propolis was collected from the whole part of Korea Peninsula, such as central, southern, and Jeju island, to analyze the general composition including total flavonoid and phenolic contents, heavy metals, colors, amino acids, and crude lipid. The total flavonoid and phenolic contents of middle region in Korea Peninsula (4.26% and 13.06 g/100 g gallic acid) were higher than southern region (2.53%, 10.9 g/100 g gallic acid) and Jeju island (0.03%, 7.22g/100 g gallic acid), respectively. The heavy metals contents showed that Zn contents were found in and a harmful heavy metal such as Cr, Cd, Pb, Ni, Cu, As, and Hg were not detected at all in our experiment. The colors of extracted propolis showed a various spectrum from yellow to red purple. Various amino acids were also detected as proline, valine, methionine and the average crude lipid contents of propolis were 42.4%. All data collected in South Korea were very first executed in our institution for the purposes of utilizing and developing the industrial insects, honeybees as well as silkworm.

Heavy metals and residual agricultural chemicals in Bombyx mori silkworm cocoon for nontextile usages were examined. Proximate composition of silkworm cocoon was examined; contents of moisture, crude protein, crude fat, and ash were about 5 ~ 6%, 93 ~ 94%, 0.4 ~ 0.5%, and 0.3 ~ 0.4%, respectively. The Heavy metals were analyzed by ICP/MS after acid digestion. Arsenic, cadmium, and lead were not detected. The average contents of mercury, zinc, and cupric were 0.317, 3.99, and 2.06 mg/kg, respectively. Agricultural chemicals, such as dichlorvos and paraquart also were not detected. Silkworm cocoon may be safe used for nontextile applications.

Cordyceps and its allies fungi has been described as a secret medicine that gives eternal youth and a long life. Some species of Cordyceps are sources of biochemicals, such ascordycepin, with interesting biological and pharmacological properties. Hence, it has been studied to uncover its pharmacological effect. We attempted to study the formation of fruiting bodies and to develop means of mass production Korean isolate of Paecilomyces tenuipes has been inoculated into silkworms, where it reproduced using culture container, mini-kit successfully. Culture container, mini-kit is composed of a cylinder-shaped body and lid. The container is made of translucent polyethylene terephthalate. The size of the container is 82 × 75 mm, reduced by 10 times as compared with the conventional culture kit. The mini kit has many advantages - high culture amount, ability of maintaining optimal humidity, parasite-free cultivation and high-end appearance. With the kit, the optimal cultivation condition is under 22 o C, culture period of 53 days. And synnemata of P. tenuipes could be kept fresh for 14 days at the temperature of under 10 o C. Therefore, the Min-kit can be used in both ways as a culture container and a packing kit for end-user customers.

The aim of this study was to evaluate the regenerative capacity of a newly developed nerve guidance conduit using electrospun silk fibroin (SFNC) implanted in a 10-mm defect of the sciatic nerve in rats. After evaluating the physical properties and cytocompatibility of SFNC in vitro, rats were randomly allocated into three groups: defect only, autograft and SFNC. To compare motor function and abnormal sensation among groups, ankle stance angle (ASA) and severity of autotomy were observed for 10 weeks after injury. Immunostaining with axonal neurofilament (NF) and myelin basic protein (MBP) antibodies were performed to investigate regenerated nerve fibres inside SFNC. ASA increased significantly in the SFNC group at 1, 7 and 10 weeks after injury compared to the defect only group (p

Bone disease can be associated with bone resorption by osteoclasts, and interest in the development of antiresorptive agents has recently increased. The hydrolysate of silk fibroin has been studied with respect to such biomedical applications. In a previous study, silk fibroin showed indirect inhibitory effects on the differentiation of osteoclasts. To further evaluate the effect of a hydrolysate of silk fibroin on osteoclasts, we investigated the direct effects of the silk fibroin hydrolysate on osteoclastogenesis and apoptosis of osteoclasts induced by receptor activation of nuclear factor κB ligand (RANKL). The silk fibroin hydrolysate inhibited RANKL-induced formation of tartrate-resistant acid phosphatase (TRAP) in RAW 264.7 cells. The inhibitory effect of the silk fibroin hydrolysate resulted in the decreased expression of osteoclast marker genes, such as matrix metalloproteinase-9 (MMP-9), cathepsin-K and calcitonin receptor (CTR). In addition, the silk fibroin hydrolysate blocked the signaling pathways of mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) and expression of transcription factors, such as nuclear factor of activated T cells c1 (NFATc1) and NF-κB. Finally, the silk fibroin hydrolysate induced apoptosis signaling cascades. Taken together, the present results indicate that silk fibroin hydrolysate has antiresorptive activity by both inhibiting osteoclastogenesis and inducing osteoclast apoptosis.

Sericultural and Agicultural Materials Division, National Academy of Agricultural Science, RDA, Suwon 441-100, KoreaReceived: November 10, 2011 / Revised: November 18, 2011 / Accepted: December 8, 2011Silk fibroin (SF) peptide has been traditionally used as atreatment for flatulence, spasms, and phlegm. In this study,we examined whether SF peptide enhanced the anti-inflammatory effect of PEP-1-FK506 binding protein(PEP-1-FK506BP) through comparing the anti-inflammatoryactivities of SF peptide and/or PEP-1-FK506BP. In thepresence or absence of SF peptide, transduction levels ofPEP-1-FK506BP into HaCaT cells and mice skin andanti-inflammatory activities of PEP-1-FK506BP wereidentified by Western blot and histological analyses. SFpeptide alone effectively reduced both mice ear edemaand the elevated levels of cyclooxygenase-2, interleukin-6and -1β, and tumor necrosis factor-α, showing similaranti-inflammatory effect to that of PEP-1-FK506BP.Furthermore, co-treatment with SF peptide and PEP-1-FK506BP exhibited more enhanced anti-inflammatoryeffects than the samples treated with SF peptides or PEP-1-FK506BP alone, suggesting the possibility that SFpeptide and PEP-1-FK506BP might interact with eachother. Moreover, the transduction data demonstrated thatSF peptide did not affect the transduction of PEP-1-FK506BP into HaCaT cells and mice skin, indicating thatthe improvement of anti-inflammatory effect of PEP-1-FK506BP was not caused by enhanced transduction ofPEP-1-FK506BP. Thus, these results suggest the possibilitythat co-treatment with SF peptide and PEP-1-FK506BP maybe exploited as a useful therapy for various inflammation-related diseases.Keywords: Inflammation, PEP-1-FK506BP, protein transduction,reactive oxygen species, silk fibroin peptide, TPADuring various cellular processes, reactive oxygen species(ROS), such as hydroxyl radicals, superoxide anion, andhydrogen peroxide, are generated and, consequently, can causelipid peroxidation, DNA damage, cell death, and variousdiseases, resulting in inflammation, cancer, Parkinson’s disease,and ischemia [1, 5, 19]. Treatment with lipopolysaccharides(LPS) or 12-O-tetradecanoylphorbol-13-actate (TPA), knownas inflammation inducers used in animal models [25, 30],can activate macrophages and secrete pro-inflammatorymediators, such as cyclooxygenase-2 (COX-2) and induciblenitric oxide synthase (iNOS), and pro-inflammatory cytokines,including interleukin-6 (IL-6), interleukin-1beta (IL-1β), andtumor necrosis factor-alpha (TNF-α). In addition, nuclearfactor-kappaB (NF- κB) and mitogen-activated protein kinase(MAPK) pathway are related to inflammatory responses[7, 8].FK506 binding proteins (FK506BPs) are members of animmunophilins family, which can bind immunosuppressivedrugs such as rapamycin and FK506, used in prevention ofgraft rejection following organ transplantation [2, 11, 21].In particular, FK506BP, a small peptide of 12 kDa, wasfirst associated with rapamycin. The resultant FK506BP-rapamycin complex binds to and then disrupts the formationof activated mammalian target of rapamycin complex 1(mTORC1), which can control protein synthesis, cellproliferation, and cell growth in response to nutrients andgrowth factors, leading to the suppression of proteinsynthesis and cell growth [4, 6, 16]. Furthermore, wepreviously reported that, through fusing FK506BP with a

A newly developed silkworm variety, GoldenSilk makes bright yellow cocoon. In this article, the basic characteristics of GoldenSilk cocoon were examined through color analysis, degumming characteristics and UV spectrophotometer. The colorant of GoldenSilk cocoon was not distributed evenly throughout the cocoon layer but existed in the outer layer of the cocoon. The colorant has not good resistance against sunlight. The degumming loss percentage of GoldenSilk cocoon was 29.2%, which is relatively higher than that of the authorized one. UV spectrophotometer showed specific absorption band around 400 to 500 nm, which is the specific yellow colorant of the GoldenSilk cocoon extracted to the degumming solution during the degumming process.

The effect of cultivation season on the cocoon filament quality of authorized silkworm variety, GoldenSilk was examined with cocoon filament character, reeling character, and fiber character during 3 years. In cocoon filament character, filament length of spring and autumn cocoon was 979 and 879 m, respectively. Cocoon filament weight of spring and autumn cocoon was 33.1 and 26.5 cg, which is lower than that of general bivoltine strain, BaeGokJam. Raw silk percentage of spring and autumn cocoon was 15.65 and 13.77%, respectively. In reeling character, reelability showed similar trend regardless of cultivation season and strain. However, the length and weight of non-broken filament of spring season were slightly higher than those of autumn cocoon. Cultivation season did not affect on the fiber characteristics such as clearness and neatness, lousiness, and degumming loss. In cocoon character, filament length, size, and weight, spring cocoon filament showed higher value than autumn cocoon, but lower than authorized general silkworm varieties. In reeling characteristics, non-broken length and weight showed the similar trends with the cocoon filament character. However, the reelability of GoldenSilk cocoon was not affected by cultivation season. The reelability of GoldenSilk cocoon was also similar with that of authorized general one. Fiber character including clearness and neatness, lousiness, and degumming loss was also similar regardless of the rearing season and different strain.

Bee venom (BV) has been used in medicine to treat a variety of diseases including arthritis, rheumatism, and various cancers. Recent reports indicate that BV has anti-angiogenic effects, but the precise molecular mechanism underlying the effects of BV against colorectal cancer remains to be elucidated. We examined the effects of BV and its major components (melittin and apamin) on tumor angiogenesis and found that BV significantly decreased protein levels of hypoxia-inducible factor- (HIF-), an important factor involved in angiogenesis and tumor progression, in human colorectal carcinoma HCT116 cells. BV also suppressed the transcription of HIF- under hypoxia, leading to a decrease in the expression of vascular endothelial growth factor (VEGF), a major target gene of HIF-. We also found that these effects were mainly elicited by apamin, but not melittin. BV specifically inhibited the phosphorylation of ERK1/2 without changing the total levels of this protein, but had no effect on kinases of p38/JNK and AKT. Our results suggest that BV may inhibit human colorectal cancer progression and angiogenesis by inhibiting HIF- and VEGF expression, thereby providing a novel potential mechanism for the anticancer action of BV.

Transparency and insolubility of eardrum patch against exudates are important for otolaryngological surgery. The author prepared silk fibroin (SF) films with various concentrations and temperature and then examined solubility and conformation of SF films. SF films were transparent regardless of the various preparation conditions. Although most SF films are soluble in 1X PBS solution at for 1 h, the SF film with 3.4% with was insoluble. Scanning electron microscopy (SEM) showed that the SF films have solid and smooth surface. Fourier transform infrared spectroscopy (FT-IR) and differential scanning calorimetry (DSC) results showed that the conformation of SF films was influenced by the preparation conditions including SF concentration and casting temperature. In conclusion, SF membrane with transparence and insolubility against exudates could be considered as eardrum patch resources.

This study was conducted to test the use of colored cocoon silkworm, Golden Silk, as a host for the fruiting body production of Cordyceps mushroom. Golden Silk variety had a shorter larval period than Yangwonjam at and 78.5% RH: The average larval period of the Golden Silk was 24 days and 7 hr, and that of the Yangwonjam was 24 days and 15 hr. In the infection test of I. tenuipes on silkworm, the larval period of normal silkworms was longer than the silkworms inoculated with I. tenuipes in both Golden Silk and Yangwonjam. The pupae survival percentage of Golden Silk was 94.4%, whereas it was 91.9% in Yangwonjam. Golden Silk had a higher pupation rate than Yangwonjam. The pupation rate of normal silkworms was about 5% higher than that of silkworms inoculated with I. tenuipes. Infection rate of I. tenuipes into larvae of 5th instar newly exuviated silkworm was 93.8% in Golden Silk, and 96.9% in Yangwonjam. Synnemata production of I. tenuipes was better in Yangwonjam with an incidence rate of 99.8% than Golden Silk with 98.6%. The average synnemata weight of Golden Silk was 1.12 g and that of Yangwonjam 1.29 g. Golden Silk had a lower synnemata weight than Yangwonjam. But no significant difference was observed in synnemata weight varieties. The synnemata of I. tenuipes produced on pupae were white or milky-white in color, and similar in shape and color to wild synnemata collected in Korea.

Summary Background Wound healing is a dynamic and complex process of tissue repair, which involves a number of cellular and molecular events. It progresses from an inflammatory response to re-epithelialisation and, finally, to the formation of a permanent scar. The pharmacological activities of honeybee ( Apis melifera L.) venom (BV) have been used in wound healing for centuries. Methods To study wound healing, full-thickness skin defects were produced on the dorsal area of mice. We measured the relative sizes and conducted histological assays of the wounds on days 3, 5 and 7. The expressions of transforming growth factor (TGF)-β1, fibronectin, vascular endothelial growth factor (VEGF) and collagen-I mRNA in the wound healing area was measured by reverse transcription polymerase chain reaction (RT-PCR). The amount of TGF-β1, fibronectin, VEGF and collagen-I was determined using immunohistochemical staining. Results The wound sizes were small in the BV group compared with the control and Vaseline groups. The BV group demonstrated decreased TGF-β1, fibronectin and VEGF mRNA levels and increased collagen-I mRNA levels. The expressions of TGF-β1, fibronectin and VEGF proteins were significantly lower in the BV group compared with the control group, while the expression of collagen-I was increased in the BV group as indicated by immunohistochemical staining. Conclusion These data suggested that BV had significant wound-healing activity. The results from this study indicated that the effects of BV on wound healing may involve biological mechanisms associated with the expressions of TGF-β1, fibronectin, VEGF and collagen-I.

Object To improve the safety of dura repair in neurosurgical procedures, a new dural material derived from silk fibroin was evaluated in a rat model with a dura mater injury. Methods The authors prepared new, transparent, artificial dura mater material using silk fibroin from the silkworm, Bombyx mori. The cytotoxic and antiinflammatory effects of the artificial dura mater were examined in vitro and in vivo by histological examination, western blotting, and reverse transcription polymerase chain reaction analyses. Results The novel artificial dura mater was not cytotoxic. However, it efficiently reduced cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase expression as well as the expression of the proinflammatory cytokines IL-1β, IL-6, and tumor necrosis factor–α. Cerebrospinal fluid leakage did not occur after repair of the brain of craniotomized rats with the artificial dura mater material. Conclusions The new artificial dura mater described in this study appears to be safe for application in neurosurgical procedures and can efficiently inhibit inflammation without side effects or CSF leakage. Although the long-term effects of this artificial dura mater material need to be validated in larger animals, the results from this study indicate that it is suitable for application in neurosurgery.

Antheraea pernyi silk fibroin (SF) hydrolysate were characterized using UV-VIS spectrometer, amino acid composition and heavy metal contents to explore its potential sources for food or cosmetic additives. The hydrolyzed A. pernyi SF was separated into two parts: (a) SFA, alanine-rich fraction and (b) SFB, tyrosine-rich fraction. SFB exhibited strong absorption peaks at 210 and 280 nm due to the presence of the tyrosine. Heavy metal analysis showed that arsenic and mercury did not detect. Other heavy metals, which includes lead, cadmium, etc., were recorded only a trace amount. Therefore, A. pernyi SF hydrolysate could be safely used as sources of food, cosmetic and pharmaceuticals.

Objective The objective of this study was to determine the capability of silk fibroin powder as a biomaterial template for the restoration of peri-implant defects when mixed with Choukroun platelet-rich fibrin (PRF) in vivo. Study design Ten New Zealand white rabbits were used for this study. Using a trephine bur (diameter 7.0 mm), 2 monocortical defects were prepared. Subsequently, 2 dental implants were installed into the tibia (diameter 3.0 mm, length 10.0 mm). In the experimental group, the peri-implant defect was filled with a combination graft of silk fibroin powder and Choukroun PRF. The control was left in an unfilled state. The animals were killed at 8 weeks. Subsequently, a removal torque test and a histomorphometric analysis were done. Results The removal torque for the experimental group was 30.34 ± 5.06 N·cm, whereas it was 21.86 ± 3.39 N·cm for the control. The difference between the 2 groups was statistically significant (P = .010). Mean new bone formation was 51.93 ± 27.90% in the experimental group and 11.67 ± 15.12% in the control (P = .003). Mean bone-to-implant contact was 43.07 ± 21.96% in the experimental group and 15.37 ± 23.84% in the control (P = .002). Conclusion A peri-implant defect can be successfully repaired by the application of Choukroun PRF and silk fibroin powder.

This study was conducted to investigate optimal conditions for formation of synnemata from Beauveria bassiana. The strain of B. bassiana was isolated from a host of Oncotympana fuscata in Halla mountain of Jeju island. The yield of synnemata was the highest at application of brown rice (90%) and silkworm pupae (10%) media. On the other hand, the other media did not result in formation of synnemata. The highest formation of synnemata was achieved at conditions of moderate air inflow and . The optimal moisture and light intensity for formation of synnemata were 60% and 300 lux, respectively. In addition, inoculation of the liquid spawn resulted in higher yield than that of asexual spore.

Honeybee (Apis mellifera) venom (BV) has a broad array of therapeutic applications in traditional medicine to treat variety of diseases. It is also known that BV possesses anti-inflammatory and anticancer effect and that it can inhibit proliferation and induces apoptosis in cancer cells, but there is no evidence of information regarding anti-apoptosis of BV on hepatocytes. In the present study, we investigated the anti-apoptotic effect of BV on tumor necrosis factor (TNF)-alpha with actinomycin (Act) D induces apoptosis in hepatocytes. TNF-alpha/Act D-treated hepatocytes were exposed to different low concentration (1, 10 and 100 ng/mL) of BV. Our results showed statistically significant inhibition in DNA damage caused by BV treatment compared to corresponding TNF-alpha/Act D-treated hepatocytes. BV suppressed TNF-alpha/Act Dtreated activation of bcl-2 family and caspase family, which resulted in inhibition of cytochrome c release and PARP cleavage. These results demonstrate that low concentration BV possess a potent suppressive effect on anti-apoptotic responses of TNF-alpha/Act D-treated hepatocytes and suggest that these compounds may contribute substantial therapeutic potential for the treatment of liver diseases.

Matrix metalloproteinase-9 (MMP-9) plays an important role in the invasion and metastasis of cancer cells. In this study, we examined the inhibitory effect of bee venom (BV) and its major peptides, melittin and apamin, on PMA-induced invasion induced by MMP-9 expression in Caki-1 renal cancer cells. BV and melittin, but not apamin, significantly suppressed PMA-induced invasion by inhibiting MMP-9 expression in Caki-1 cells. Furthermore, as evidenced by MMP-9 promoter assays, melittin inhibited MMP-9 gene expression by blocking the PMA-stimulated activations of activator protein-1 (AP-1) and nuclear factor-kappa B (NF-kappaB). In addition, melittin suppressed the PMA-induced phosphorylations of ERK and JNK mitogen-activated protein kinases, upstream factors involved in Ap-1 and NF-kappaB. These results suggest that the suppression of MMP-9 expression contributes to the anti-tumor properties of melittin.

Bee venom (BV) has a long tradition of use for the control of pain and inflammation in various chronic diseases. Carbon tetrachloride ( CCl4) is known to induce hepatotoxicity after being metabolized to the highly reactive trichloromethyl free radical and its peroxy radical. The purpose of the current study was to examine whether BV regulates the pro-inflammation and fibrosis related genes against a mouse model of hepatic fibrosis induced by CCl4and ethanol-treated hepatocytes (ETH). Test mice were administered with CCl4(2 ml/mg) and hepatocytes were treated with 25 mM ethanol. BV was added to the final concentration of 0.05–0.5 mg/kg and 1–100 ng/ml for in vivo and in vitro testing, respectively. Fibrotic livers and ETH were used for the measurement of hepatocyte necrosis, pro-inflammatory cytokines and fibrogenic genes. BV suppressed CCl4-induced hepatocyte necrosis markers of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT). It also inhibited the secretion of interleukin (IL)-1β and tumor necrosis factor (TNF)-α. Moreover, BV inhibited CCl4-induced expression of transforming growth factor (TGF)-β1, α-smooth muscle actin (SMA) and fibronectin. Similarly, ETH exhibited significant suppression of IL-1β, TNF-α, TGF-β1 and fibronectin when cultured with BV. These results suggest that BV possesses anti-fibrogenic properties that are mediated by the suppression of pro-inflammatory cytokines and fibrogenic gene expression. BV has substantial therapeutic potential for the treatment of fibrotic diseases.

Bee venom (BV) has been used as treatment against a wide variety of ailments, including inflammatory diseases. Various studies have demonstrated anti-inflammatory and anticancer effects of BV. Transforming growth factor (TGF)–β1 induces hepatocyte apoptosis via the mitochondrial permeability transition. However, there is no evidence or information regarding the antiapoptotic effect of BV on hepatocytes. The authors investigated the antiapoptotic effect of BV on TGF-β1-treated hepatocytes. The results showed significant protection from DNA damage by BV treatment compared to corresponding TGF-β1-treated hepatocytes without BV. BV suppressed TGF-β1-induced activation of the bcl-2 family and caspase family of proteins, which resulted in inhibition of poly ADP-ribose polymerase (PARP) cleavage. Furthermore, BV is not cytotoxic in the low concentrations used in this study. Low concentrations of BV potently suppress the apoptotic response in TGF-β1-treated hepatocytes; therefore, BV may have therapeutic potential for the treatment of liver diseases.

The components of bee venom (BV) utilized in the current study were carefully scrutinized with chromatography. Despite its well documented anti-inflammatory property, there are no reports regarding the influence of BV on the expression of cellular adhesion molecules in the vascular endothelium. A great amount of information exists concerning the effects of an atherogenic diet on atherosclerotic changes in the aorta, but little is known about the molecular mechanisms and the levels of gene regulation involved in the anti-inflammatory process induced by BV. The experimental atherosclerosis was induced in mice by a lipopolysaccharide (LPS) injection and an atherogenic diet. The animals were divided into three groups, the NC groups of animals that were fed with a normal diet, the LPS/fat group was fed with the atherogenic diet and received intraperitoneal injections of LPS, and the LPS/fat + BV group was given LPS, an atherogenic diet and intraperitoneal BV injections. At the end of each treatment period, the LPS/fat + BV group had decreased levels of total cholesterol (TC) and triglyceride (TG) in their serum, compared to the LPS/fat group. The LPS/fat group had significant expression of tumor necrosis factor (TNF)-α and interleukin (IL)-1β in the serum, compared with the NC group ( p < 0.05). The amount of cytokines reduced consistently in the BV treatment groups compared with those in LPS/fat group. BV significantly reduced the amount of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), transforming growth factor-β1 (TGF-β1) and fibronectin in the aorta, compared with the LPS/fat group ( p < 0.05). A similar pattern was also observed in the heart. In conclusion, BV has anti-atherogenic properties via its lipid-lowering and anti-inflammatory mechanisms.

This study examined the effect of whole bee venom (BV) as a potential stimulant of the piglet immune system, on growth performance, blood parameters, plasma protein and immune globulin content of serum. Piglets (n = 97) received combinations of 0.5, 1.0, 1.5, 2.0 and 2.5 mg/kg of parenterally administered BV on 4 occasions between birth and Day 30. In the apipuncture group (n = 31), piglets were acupunctured with the worker honeybee. Two acupoints, GV-1 (Jiao-chao) and GV-20 (Bai-hui), were selected for apipuncture. All piglets (n = 128) in the treatment groups were treated 4 times throughout the study period of 60 days. The control piglets received no treatments. Blood was taken via jugular venipuncture on Day 30 after birth. Body weight and survivability were measured, and changes in hematological values were analyzed. Both the BV injection group and apipuncture group increased body weight and survivability by 26.6% and 21.8%, and 7.9% and 6.7% respectively compared to the controls. The numbers of leukocytes, erythrocytes, lymphocytes and monocytes were not influenced by treatments. However, a potential clinical benefit of high dose therapy was seen in increased populations of leukocytes, lymphocytes and monocytes compared with either the apipuncture or control groups. Other blood parameters such as total protein and albumin were not affected by treatment. However, IgG levels were generally higher in treated groups than in the controls. These findings indicate that BV might be useful to stimulate immuno-competence in pig production, possibly via the primary bioactive components of melittin, phospholipase A 2 and apamin. The administration of BV, either via injection or acupuncture, did not make any differences in growth performance of young pigs. These results would be useful for further purification and characterization of immune boosting agents from BV.

In this study, we investigated the indirect effect of silk-fibroin on osteoclastic differentiation of RAW264.7 cells. The conditioned medium were collected from MC3T3-E1 osbeoblasts treated with silk fibroin for 6 days, mixed in 1:1 ratio with osteoclast medium, and then added into RAW264.7 cells with receptor activator of nuclear factor kappa B ligand (RANKL), a differentiation inducer for 3 days. Of osteoclastic cytokines in the conditioned medium, the protein expression of osteoprotegerin (OPG) with silk-fibroin was not significantly different. However, the protein expression of interleukin (IL)- was specifically lower in a dose dependent manner. In RAW264.7 cells, the conditioned medium with silk-fibroin inhibited RANKL induced osteoclastic differentiation as total number of multinucleated tartrate-resistant alkaline phosphatase (TRAP)-positive osteoclasts in a dose dependent manner. Taken together, we demonstrated that the conditioned medium of silk-fibroin treated osteoblasts inhibits RANKL induced differentiation of osteoclasts with inhibiting selective expression of IL-.

Pure Bombus ignitus venom samples were submitted to two-dimensional gel electrophoresis. A total of 64 excised spots were analyzed by mass spectrometry. Three main proteins resulted in the identification have not been described in other bee venoms before. Dose-dependence against human carcinoma (Hep3B, BT-20, A549 and AGS) were observed from 1ng/ml to 100ng/ml. Expecially, the treatment of 100ng/ml B. ignitus venoms showed the highest cytotoxicity with 55% against hepatocellular carcinoma (Hep3B). The B. ignitus venoms showed strong antimicrobial activities against Enterococcus faecium and Shigella sonnei, and practically antimicrobial activity against the other microorganisms tested. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of E. faecium and S. sonnei, were 0.256ug/ml, respectively.