Earticle

To treat hyperpigmentating diseases such as such as melasma and lentigines, a number of melanogenesis inhibitors have been screened for their effectiveness in reducing melanin but none have shown to be completely satisfactory. To develop a novel skin depigmenting agent from natural sources, the inhibition of melanogenesis by Chinese plant, N. glandulifrea, was evaluated. The methanol extract of this plant showed significantly down-regulated melanin synthesis in a dose dependent manner at a non-toxic concentration in cultured B16F10 mouse melanoma cells. This extract was further fractionated by using bioactivity-guided fractionation and identified as glycerol monolinoleate by means of NMR, IR and HPLC. This compound showed significant depigmenting activity melan-a cells. Compared with arbutin (IC50 = 0.74 mM) as a positive control, the depigmentation IC50 value for this compound was 9.1 mM. Its inhibitory effect on cellular tyrosinase, the key enzyme of melanogenesis, was observed. The effect of this compound on tyrosinase and MITF protein was also investigated. These results suggest that glycerol monolinolate isolated from N. glandulifera is a promising compound that could be useful for treating hyperpigmentation as a skin-whitening agent.

Silkworm silk is composed of fibroin and sericin and its hydrolysate has been known to be an excellent skin moisturizer, wrinkle controller, and anti-oxidant. The silk proteins, fibroin and sericin contain rich in glycine and serine among 18 different amino acids. Since glycine and serine are the amino acids that consist of mor than 40% of skin amino acids, the silk hydrolysate has been widely used as raw material for cosmetics and functional food. In this study, silk hydrolysate prepared from silkworm gland in Bombyx mori by treating with hot alkali solution was analyzed and compared with that from silkworm cocoon. Compared to silkworm cocoon hydrolysate, the silkworm grand hydrolysate was 15 times more viscose at neutral pH and showed much better skin affinity. The amino acid composition and heavy metal content showed no significant differnce in both hydrolysates but the molecular weight distribution analyzed by SDS-PAGE demonstrated that the molecular weight of silkworm grand hydrolysate is relatively lower than that of silkworm cocoon hydrolysate, which suggests that silkworm gland is more susceptible to alkali hydroysis. From this comparative analysis, the silkworm grand hydrolysate looks very promising if used as raw material for cosmetics.

The inhibitory effect on melanogenesis was studied to develop a new natural whitening agent for cosmetics. Various extracts of Clematis apiifolia DC. were tested for the screening. Free radical scavenging activities of extracts (DPPH) IC50 were analyzed for the antioxidant capability of extracts. Mushroom tyrosinase inhibition activities were tested for the enzymatic melanogenesis inhibition. Further inhibitory effect on melanogenesis was tested with B16 melanoma cells by measuring the content of melanin synthesized. Clematis apiifolia DC. was resulted as a good candidate of a new natural whitening agent for cosmetics.

The flathead mullet, Mugil cephalus, is a mullet of the genus Mugil, found in coastal tropicaland subtropical waters worldwide. The flathead mullet is an important food fish for many around the world, and can be both fished and farmed. The roe of this mullet is salted, dried, and compressed to make a specialty food across the world, such as Korean myeongran jeot. In this study, amino acid contents of hot-water extracts of Mugil cephalus were analyzed for cosmetic application. Several cosmetic formulations have been prepared to measure moisturizing and soothing effects on skin. The moisturizing effect increased with the extracts concentration. Some advantages on the skin effect of the extracts were discussed also. This research was financially supported by the Ministry of Education, Science Technology (MEST) and Korea Institute for Advancement of Technology(KIAT) through the Human Resource Training Project for Regional Innovation(2010)

Vitiligo is an acquired pigmentary disorder characterized by areas of depigmented skin resulting from loss of epidermal melanocytes. The prevalence of this disease varies from 0.1% to 2% in various global populations. This is one of the most psychologically devastating skin diseases. Histologically, the predominant finding in the depigmented areas of vitiligo is an absence of epidermal melanocytes. The precise cause of the loss of these epidermal melanocytes is unknown. Autoimmune, neural, biochemical, oxidative stress, autocytotoxic, viral, and melanocytes detachment mechanisms have been proposed to explain the pathogenesis of vitiligo. It is known that the existance of inactive melanoblasts in hair root follicles provide a melanocyte source for repigmentation in Vitiligo. Differentiation and migration of these melanocytes precursors from the outer root sheath of hair follicles into clinically depigmented epidermis is crucial for the repigmentation in Vitiligo therapy. An effective treatment regime has been proposed that would stimulate the inactive melanoblasts in the hair follicle to divide, proliferate and migrate upward along the surface of outer root sheath to the nearby epidermis. To evaluate the potential of herbal candidates in treatment of Vitiligo, we studied its effects on the differentiation, proliferation and migration of melanoblast cellsScreening of natural compounds were carried out by evaluating the effect of plant extracts to induce differentiation of melanoblast to active melanocytes by accessing the melanin production (melanin assay) and proliferation (by MTT assay) in melanoblast cell lines (Melb-a).

MITF (microphthalmia-associated transcription factor), a bHLH (basic helix-loop-helix) transcription factor with a leucine zipper, is a key regulatory protein for the pigmentation process in melanocytes. Binding of MITF to E-box caused transcription of several pigmenting genes including the tyrosinase gene. The purpose of this study is to analyze the interaction of MITF and its DNA binding site (E-box, CATGTG) by surface plasmon resonance. Label free DNA-protein interaction assay, SPR (surface plasmon resonance) was used to analyze their interactions on the gold surface. It is expected that SPR can be a useful method to study of MITF and E-box interactions. And we hope to apply this study as a screening tool for depigmenting agents in the cosmetic industry.

Skin color is determined by the amount of melanin present. Many enzymes have a relationship in melanin synthesis. Especially, tyrosinase is well known as key enzyme for melanin synthesis. MITF (Microphthalmia-associated transcription factor) belongs to the basic helix-loop-helix-zip family of transcription factors and is the major regulator of tyrosinase and other related enzymes (TRPs) by binding E-box (CATGTG). To screen for MITF – E-box binding inhibitor, EMSA (Electrophoretic Mobility Shift Assay) was performed. Based on the computer-simulated structure, 27 chemicals were selected and were investigated as MITF – E-box binding inhibitors. To check the depigmenting activity, cell tests were performed by using melan-a cells. Among them, compound #18 was found to show the most potent inhibitory activity against MITF – E-box binding. As a result of EMSA, intensities of MITF – E-box bands were reduced by compound #18 in a dose-dependent manner. In addition, unlabelled Tyr-p, mutated Tyr-p and random oligomer were used for EMSA to investigate the competitive binding inhibitory effect. As a result, excess unlabelled Tyr-p was an effective binding competitor. Conversely, mutated Tyr-p and random oligomer did not compete with MITF. To check the protein level of MITF, tyrosinase, and TRP-1, western blot analysis was performed by using melan-a cells. The protein level of tyrosinase was reduced by compound #18, but MITF and TRP-1 were not reduced. Therefore, compound #18 could be used as a specific MITF – E-box binding inhibitor on melanocytes.

Fucoidan is a marine sulphated polysaccharides of cell-wall matrix of brown algae, containing large proportions of L-fucose and sulphate, together with minor amounts of other sugars like xylose, galactose, glucose, mannose, uronic acid, and rhamnose. For the past years, fucoidan has been extensively studied due to its numerous biological activities such as anticancer, anticoagulant, antithrombotic, antiinflammatory and antiviral. However, the whitening activity of fucoidan has not been elucidated yet. This study was to identify the depigmenting effect of crude fucoidan isolated from the sporophyll of Korean brown seaweeds Undaria pinnatifida at coastal area of Wando, South Korea. The crude fucoidan was isolated by acidic extraction method. Melanin assay and MTT assay was done to confirm the depigmenting activity in B16F10 melanoma cells and melan-a melanocyte cells. The results showed that the crude fuicoidan exhibit 29% melanin inhibition in B16F10 melanoma cells and 54% melanin inhibition in melan-a melanocyte cells at 100 ㎍/㎖ . In the both cell lines, crude fuicoidan did not showed cell cytotoxicity up to the concentration 100 ㎍/㎖. The crude fucoidan has been purified to determine its structure.

Coenzyme Q10 (CoQ10) is a fat-soluble, vitamin-like substance found in the cells of many organisms. Primary role of CoQ10 is in the transfer of electrons from membrane-bound dehydrogenases to complex III of the electron transport chain. CoQ10 also acts as an antioxidant. It is used as a nutritional supplement and is beneficial in the treatment of human conditions, such as cardiomyopathy, diabetes, Parkinson’s and Alzheimer’s disease, and can also reduce the risks of myopathy associated with the use of statin drugs. The use of CoQ10 is also gaining popularity in the cosmetic industry owing to its antioxidant properties. In spite of the versatility of CoQ10, the molecule's long side chain of 10 isoprenoid units make CoQ10 poorly water-soluble, limiting its bioavailability and more various applications. In the present study, a novel CoQ10 nanoparticle was assembled by encircling isoprenyl chains with human apolipoprotein A-I protein (apoA-I). This nanostructure of CoQ10 is quite reminiscent of nascent high density lipoprotein (HDL) particle composed of phospholipids and cholesterol. The diameter of the CoQ10 nanostructure was about 12 nm, which is less than 1/10~1/100 of liposome-based CoQ10 particles. Novel CoQ10 nanostructure will be versatile in pharmaceutics and functional cosmetics. We expect that the strategy suggested in this study can be exploited to assemble nano-sized, water-soluble structures of various water-insoluble drug candidates (i.g. Paclitaxel).

Effective drug delivery to a target site while minimizing drug exposure elsewhere remains a difficult challenge. Major obstacles in local drug delivery include the ability to administer therapeutic doses to the target tissue, as well as noninvasive measurement of the concentration distribution of administered agents to achieve the former. This study describes the development of a noninvasive in vitro magnetic resonance imaging (MRI) method to quantitatively study the distribution of drug surrogate and to measure fluid flow velocity in a region of interest (ROI). Dynamic contrast enhanced MRI was used to study diffusion-convection transport phenomena of a magnetic resonance contrast agent, gadolinium-diathlenetriaminopentaacetic acid(Gd-DTPA), in 1% agarose gel. The relationship between the concentration of Gd-DTPA and T1 relaxation time was determined using an inversion recovery MRI technique. The concentration distribution of Gd-DTPA images was estimated from a calibration curve relating T1 relaxation time and the concentration of Gd-DTPA. Using the estimated concentration profiles, center-of-mass points were calculated in a series of time points in order to determine fluid flow velocity, which correlated well with the real volumetric flow velocity at early time points. The calculated fluid flow velocity deviated from the real volumetric flow velocity calculated by the center-of-mass started deviating from the real volumetric flow velocity when the applied tracer started leaving the gel with water. In this study, we developed a method to analyze MR images quantitatively and to determine fluid flow velocity through a tissue in vivo.

Despite the recent research interest in the field of nanoparticles delivery system, their structure modification and transport behavior of various hydrophobic drugs is poorly developed. In this article the synthesis of novel amphiphilic diblock copolymer poly([R]-3-hydroxyvalerate)-block-monomethoxy poly(ethylene glycol) (PHV-blockmPEG) was undertaken by modifying the structure of biodegradable and hydrophobic poly([R]-3-hydroxyvalerate) (PHV) with hydrophilic monomethoxy poly(ethylene glycol) (mPEG). The chemical combination of the two blocks was carried out in the melt using bis(2-ethylhexanoate) tin as transesterification catalyst. The synthesized product was characterized by gel permeation chromatography (GPC), 1H nuclear magnetic resonance (NMR) and differential scanning calorimetry (DSC) analysis. The block copolymer self-assembled into amphiphilic nanoparticles with a core of hydrophobic PHV and a shell of hydrophilic mPEG in aqueous solution. Characterization of the nanoparticles showed the formation of discrete, spherically shaped nanoparticles with mean particle size of 200±1 nm and zeta potential of-14±1mV. A hydrophobic drug thymoquinone was efficiently incorporated into the core hydrophobic domain of the nanoparticles and its release kinetics was studied in vitro. The amphiphilic PEGylated nanoparticles showed biocompatibility when checked in neuronal hippocampal cells of prenatal rat. Our results suggest that the amphiphilic nanoparticles with core–shell structures are potentially useful to develop novel drug carriers.

The nano fibers were prepared from biocompatible polymers dissolved in distilled water by electrospinning. Electrospinning of biocompatible polymer was performed by blending with poly(vinyl alcohol) and poly(ethylene oxide). Chitosan and chondroitin sulfate sodium salt was used as biocompatible polymers with the biological characteristics. Water soluble polymer was mixed with biocompatible polymer by dissolving in distilled water. Various nano fibers were obtained by electrospinning at various conditions. Prepared nano fibers were compared by SEM and cytotoxicity test.

Glaucoma is a secondary irreversible blindness disease in the world.1) Especially, glaucoma mainly caused by high intra-ocular pressure(IOP) that damage to optic nerves, result in blindness. To control the IOP, various methods have been devoloped during about 100 years, such as medications, surgery and drainage device implantation2). Although all of these method have lots of benefit to treat and control the glaucoma disease, cannot to control the IOP in acute glaucoma easily, because these method developed by only focusing on the chronic glaucoma. The failure of control of IOP in the acute glaucoma can cause a drastic blindness because ocular pressure increased rapidly. Hence, an innovative device demanded to control the IOP in acute glaucoma. Here, we fabricate the metallic hollow microneedle with ultra high-aspect-ratio using drawing lithography technology3) to puncture the sclera with minimally surgery. This metallic hollow microneedle succeeded to control IOP in the glaucoma animal model. We anticipate that this metallic hollow microneedle can be applied for any other ocular disease.

Eicosapentaenoic acid(EPA) in one of omega-3 fatty acids in unsaturated fatty acids and plays a pivotal role in protection against oxidative damage-related diseases due to anti-inflammatory effects.In this study, anti-inflammatory effects of EPA loaded nano-particles(ENP) on lipopolysaccharide(LPS)-induced murine macrophage cells (RAW264.7) were investigated. The nanoparticles released EPA which was able to inhibit the production of nitric oxide (NO) by suppressing the expression of inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-activated RAW 264.7 macrophage cells. NO production and iNOS expression was significantly decreased in dose-dependent manner when cells were treated with various concentrations of ENP.

The objective of the present work was to develop a new drug nanocarrier consisting of nanoparticles made of glycol chitosan (Gc) and eicosapentaenoic acid (EPA). A primary amine of Gc was modified with EPA to prepare a hydrophobically modified Gc (EPA-Gc). The modified chitosan formed self-aggregates in an aqueous phase by intermolecular association between hydrophobic EPA attached to glycol chitosan. We have found that EPA-Gc nanoparticles can deliver multiple poorly water-soluble drugs. EPA-Gc nanoparticles were characterized in terms of size, zeta- potential, stability and drug release. The result showed that these chitosan derivatives were able to self-assemble and form spherical shape polymeric nanoparticle with the size about 90–580nm and zeta potential range of 13.94-20.33mV, depending on concentration of EPA. The size of EPA-loaded EPA-Gc nanocarrier was reduced with increasing the loading efficiency of EPA, due to the enhanced hydrophobic interaction between hydrophobic drug and hydrophobic segment of EPA-Gc. This study presents a new way to nano carrier long chain polyunsaturated fatty acids like EPA, useful for enrichment of hydrophobic drugs for delivery and release properties.

In this study, selenium nanoparticles were synthesized through the redox system. Generally, selenium nanoparticles were reported a novel selenium species with a novel biological activity and low toxicity. In the present study, we demonstrated a simple method for synthesis of size-controlled selenium nanoparticles by adding sulfated polysaccharides or sulfated amino acids to the redox system of selenite and ascorbic acid. Selenium nanoparticles were synethesized from sulfated polysaccharides such as fucoidan, ι-carrageenan, λ-carrageenan, κ-carrageenan, chondroitin sulfate sodium salt and sulfated amino acids such as L-cysteine, L-methionine. The size of selenium nanoparticles was range of 30-150nm and the average was 100nm. The size of particles decrease according as sulfated polysaccharides concentration increase from 0.1% to 3%. And selenium nanoparticles were performed the selenium content experiment and antioxidant test. The obtained products were very stable and would be used as medicine and food material.

Hydrogels have been broadly used for a variety of biomedical applications including wound-healing bioadhesives, artificial kidney membranes, articular cartilage, organ reconstruction material, and artificial skin. Natural polymer hydrogel were prepared and their properties were determined in order to select the most appropriate setting method, drug delivery material. The hydrogels were made from gellan gum, xanthan gum, curdlan, chitosan and cross-link agent. Hydrogels obtained were characterized in terms of gel content, viscosity, swelling ratio, sol-gel transition temperature, mechanical and thermal properties maximum stress(MS), tensile strength(TS), elongation(E), differential scanning calorimetry analysis(DSC). The resultant hydrogels were flexible and elastic in the dry state. Hydrogel added cross-link agent showed higher gel content, viscosity and tensile strength than hyrdogel without cross-link agent. The in vitro degradation tests showed delayed swellings properties of the hydrogels.

Development for methods identifying effective drug targets and antibacterial is urgently needed because microbial pathogens are constantly developing resistance to currently available antibiotics. Such trend has been threatening mankind, especially those with compromised immune system. We employed genome-scale metabolic modeling as it has proved to be very useful in capturing the physiological characteristics of various organisms. In this metabolic modeling, two systematic approaches, namely constraints-based flux analysis and chokepoint analysis were applied to genome-scale metabolic networks of Escherichia coli, Helicobacter pylori, Mycobacterium tuberculosis and Staphylococcus aureus; the former is an optimization- based simulation technique that calculates metabolic fluxes, while the latter is networktopology-based method that selects enzymes or metabolites as a target that has a single ingoing and/or outgoing reaction. We combined these methods to generate novel drug targets in the aforementioned emerging drug-resistant pathogens. This study demonstrates that drug targeting using in silico approaches enables a rational design of experiments applicable to biomedical science. [This work was supported by the Korean Systems Biology Research Project (20100002164) and WCU (World Class University) program (R322009000101420) of the Ministry of Education, Science and Technology (MEST) through the National Research Foundation of Korea.]

30Kc19, a 30 kDa-sized protein derived from silkworm hemolymph has been recently reported for its capability of penetrating the cellular membrane of animal cells in our group. We believe this is because 30Kc19 protein may encompass the protein transduction domain (PTD) within its sequence. PTD mediates the uptake of biologically active molecule into mammalian cells and many researchers have endeavored to deliver the therapeutic drugs into cells of living organism by using PTDs. In this study, in order verify its potential use as a tool for delivering drugs, we have managed to deliver 30Kc19 protein in vivo. Following the intraperitoneal injection of 30Kc19 into the mice, the internalization of 30Kc19 into various organs was examined by immunohistochemistry. Furthermore, not only 30Kc19 protein but also GFP fused 30Kc19 protein was delivered successfully in various organs. Through this, we have confirmed that 30Kc19 protein has the ability to mediate the uptake other fused biological molecules. Therefore, we have verified the potential use of 30Kc19 protein in efficiently delivering in vivo system, and anticipate its use as a tool for efficient drug delivery in the future.

Cancer is one of the prime causes of all human deaths in the world. Although there are many anti-cancer drug including chemotherapy to treat cancer, anti-cancer drugs are known to cause adverse effects such as injury of immune system, which constrains their use in tumor treatment. Therefore, it is necessary to investigate novel anti-tumor drugs with improving immunity without harming the host. Mushrooms have attracted more attention recently in the biochemical and medical areas due to their useful therapeutic effects. Especially, Inonotus obliquus, a traditional medicinal mushroom, has been widely used to promote health and longevity. In this study, we investigated the in vitro and in vivo immunostimulating activities of polysaccharides isolated from fruiting body of Inonotus obliquus (PFIO) on macrophages, T lymphocytes and experimental mouse model. Additionally, the signaling pathway of PFIO-mediated macrophage activation was investigated in RAW264.7 macrophage cell line.

The PGSS (Particles from Gas Saturated Solutions) process is a promising technique that uses supercritical fluid to produce fine particles under relatively mild operating conditions. In this technique, a supercritical fluid is dissolved into a solution or a melted material. Under the appropriate pressure and temperature conditions, this mixture is rapidly depressurized through a nozzle, resulting in the formation of fine solid particles. Gemcitabine is a pyrimidine nucleoside analogue with a high antitumor efficacy for the treatment of ovarian, breast, bladder, and pancreatic cancers. In this study, gemcitabine-loaded polymer particles were parepared using a PGSS technique. The effect of temperature, pressure, the type of biodegradable polymers, and micronization of gemcitabine using a supercritical ASES (Aerosol Solvent Extraction System) process on the morphology and drug entrapment efficiency of PGSS-processed particles were investigated. The entrapment efficiency of gemcitabine from particles was also determined using HPLC assay.

Cardiovascular disease is generally caused by a blockage of blood flow through heart or brain. The most common cause for blockage is a build-up of fatty deposits, inflammation and atherosclerotic plaques on the inner walls of blood vessels. It is assumed that cardiovascular disease related proteins would be secreted from the deposits or plaque on the inner wall of blood vessels. If it is possible to monitor the increase or decrease of these proteins, the incident of cardiovascular disease can be prevented. In this study, to identify the changed proteins for diagnosis of cardiovascular disease, protein profiles of human sera 20 of non-diabetic cardiovascular disease patients were compared with those of 30 healthy controls using the two-dimensional gel electrophoresis technique. Before 2-DE, high abundant proteins in serum were removed using the multiple affinity removal column connected with HPLC system. The differentially expressed spots in cardiovascular disease patients were identified by ESI-Q-TOF mass spectrometry. The identified proteins were confirmed via Western blot analysis. Among the identified proteins, RBP4 and Celluloplasmin were confirmed to be good candidates for diagnostic and monitoring biomarkers of cardiovascular disease.

Type 2 diabetes (T2D) is characterized by hyperglycemia, which is caused by insulin resistance and relative insulin deficiency. Type 2 diabetic patients have higher risk of acute liver failure than non-diabetic patients. In this study, we investigated the possible hepatic protective effects of vitamin A, as an antioxidant, in the T2D rat model. Male Sprague-Dawley rats (150-160 g, n=21) were divided into 3 groups: normal group, diabetic group and vitamin A-treated diabetic group. To confirm liver injury before and after vitamin A treatment, serum levels of hepatic enzyme markers (GOT and GPT) and the liver-to-body weight ratio were measured for 4 weeks. As a result of these experiments, we found that vitamin A was effective in protecting the T2D rat model from liver injury. Using two-dimensional gel electrophoresis (2-DE), we have compared differences of rat liver proteome. Differentially expressed proteins in the diabetes group compared to the normal group and in the vitamin A-treated diabetes group compared to the diabetic group were identified using ESI-Q-TOF MS/MS analyses. Therefore, these proteins may play a key role in the development and recovery of liver injury by vitamin A-treatment in the T2D rat model.

Thiazolidinedione analogue, CT-8, has been reported to be potent inhibitor of human 15-hydroxyprostaglandin dehydrogenase (15-PGDH). In continuing attempts to develop highly potent 15-PGDH inhibitors, a series of thiazolidinedione analogues with different substituents on phenyl rings were synthesized and tested. Of the compounds produced, those in the low nanomolar range were most effective. In particular, introduction of halogen atoms at the phenyl ring of CT-8 greatly improved its inhibitory efficacy. For structure-activity analysis, the effects of adding cyclohexyl with different chain lengths to the hydroxyl groups of 5-(3-substituted-4-hydroxybenzylidene)thiazolidine-2,4-dione were tested. Removal of the methylene group between the cyclohexyl ring and oxygen of 5-(3-substituted-4-(2-cyclohexylethoxy)benzylidene)thiazolidine-2,4-dione decreased its inhibitory potency. It appears that two methylene groups between the cyclohexyl ring and oxygen of 5-(3-substituted-4-hydroxybenzylidene)thiazolidine-2,4-dione are optimal for inhibitory activity. The most potent inhibitor of this series of compounds is compound 5, (5-(3-chloro-4-(2-cyclohexylethoxy)benzyl)thiazolidine-2, 4-dione), with an IC50 of 0.008 μM.

Recent studies have suggested the existence of a small subset of cancer cells called cancer stem cells (CSCs), which possess the ability to initiate malignancies, promote tumor formation, drive metastasis, and evade conventional chemotherapies. Elucidation of the specific signaling pathway and mechanism underlying the action of CSCs might improve the efficacy of cancer treatments. In this study, we analyzed differentially expressed proteins between tumerigenic and non-tumorigenic cells isolated from the human hepatocellular carcinoma (HCC) cell line, Huh7, via proteomic analysis to identify proteins correlated with specific features of CSCs. Among the identified proteins, COP9 and Transgelin were confirmed with RT-PCR and Western blotting. COP9, lowly expressed in CD133+ cells compared to CD133- cells, was known one of the Rb regulators. However, Transgelin related to cell migration was highly expressed in CD133+ cells. We observed CD133+ cells have more metastatic phenotype than CD133- cells using wound healing assay and invasion assay. Moreover, in tumors derived from Huh7-induced xenografts, Transgelin was also co-expressed with CXCR4, responsible for tumor invasion. More studies are needed to reveal the role of the identified proteins in the undifferentiated tumerigenic cells.

Choroidal neovascularization (CNV) is the primary cause of visual defect for the most of patients severely suffering from age-related macular degeneration (AMD). Recent research on drug delivery to the choroid where CNV originates demonstrates that has limitation on a drug penetrated into the outer retina. The objective of this study was to improve a drug delivery system for an integrin-antagonist peptide to the sub-retinal region. We developed nanoparticles that were fabricated by blending polylactic acid/polylactic acid-polyethylene oxide(PLA/PLA-PEO), and water-soluble integrin-antagonist peptide, C16Y, were encapsulated in nanoparticles(C16Y-NP). The PLA/PLA-PEO nanoparticles were 302 ± 85. 1-sized and continuously released C16Y encapsulated into NPs for over 2 weeks in in vitro. The nanoparticles did not exhibit any retinal toxicity as examined by histopathology. Either C16Y-NPs or C16Y peptide solution were separately injected to rodent models 5 or 9 days post laser photocoagulation. Both intravitreal injections of C16Y peptide and C16Y-NPs statistically suppressed CNV (p<0.05) at 5days and 9 days post laser photocoagulation. In the case of C16Y-NPs injection on day 12, however, the area of choroidal neovascularization on the day 12 was smaller than single injection of C16Y peptide solution (p<0.05) due to the short half-life in vitreous body. The result in this paper indicates that sustained release type of drug delivery is important to treat CNV derived from AMD. The intravitreally administered PLA/PLA-PEO nanoparticles encapsulating coumarin were revealed to penetrate the retina and localize to the RPE. These results propose that nanoparticles fabricated by biodegradable polymers are promising materials as carrier and potential drug delivery system may be established for treatment of AMD.

Spirulina is a blue-green microalgae which is known to be safe food and pharmaceutical additive. There are many reports that Spiulina cell mass has functional ingredients such as anti-inflammatory, anti-metastasis and blood cholesterol reduction. Electrospinning is a novel technique to fabricate nanocale fiber mats which are porous mateials with high ratio of surface area to volume. Nanofiber matrix is suitable for cell adhesion, spreading and growth. Polycaprolactone (PCL),a biodegradable and biocompatible synthetic polymer, known to fabricate good nanofibers in electrospinning. In this result, we suggest an electrospun PCL-Spirulina nanofiber as an extracellular matrix for cell culture. Acknowledgement This work was supported by INHA UNIVERSITY Reasearch Grant. (INHA-40875)

The bioequivalence of two oral formulations containing chlorphenesin carbamate 250 mg was determined in 26 healthy volunteers. The study was designed as 2 doses, fasting, two-period two-sequence crossover study with a washout period of 1 week. The content of chlorphenesin carbamate in human plasma was determined by a validated LC-MS/MS method. The preparations were compared using the parameters area under the plasma concentration time curve (AUCt), peak plasma concentration (Cmax), and time to reach peak plasma concentration (Tmax). No statistically significant difference was observed between the logarithmic transformed AUCt and Cmax values of the two preparations. The 90% confidence interval for the ratio of the logarithmic transformed AUCt and Cmax were within the bioequivalence limit of 0.80~1.25.

Anti-angiogenesis was carried out for investigating of anti-cancer and anti-obesity mechanism from natural products such as Anthrisci Radix, Psoraleae Semen. The formation of angiogenesis was completely inhibited by these 2 kinds of natural products. The maximum safe dose of Anthrisci Radix, Psoraleae Semen is 0.2 ppm and 10 ppm, and the fraction of these natural products also showed higher anti-angiogenic effect, F5 and F2 fraction, respectively. F5 fraction of Anthrisci Radix completely surppressed angiogenesis with incresing its dose, while F2 fraction of Psoraleae Semen inhibited angiogenesis at 10 ppm. These 2 kind of natural products will be a good anti-angiogenic substances for anti-cancer and anti-obesity.

The thiol-disulfide oxidoreductase thioredoxin-1 (TRX) is known to be secreted by leukocytes and to exhibit cytokine-like properties. Extracellular effects of TRX require a functional active site, suggesting a redox-based mechanism of action. However, specific cell surface proteins and pathways coupling extracellular TRX redox activity to cellular responses have not been identified so far. We first found that exogenous TRX not only scavenger but also effected intracellular proteins using proteomic approaches. Therefore, the effect of a thiol-antioxidant protein TRX have appropriated on regulation in a human skin malignant melanoma cell line (A375). Mechanism-based His pull-down and immunoprecipitation technique used to identify TRX binding partner proteins. We found that up/down regulated intracellular proteins were identified under the condition of LPS-treated A375 cells and co-treat with exogenous TRX using in vitro 2-DE proteomic approaches. As well as, confirmed in vivo mouse inflammation models used to quantitatively measure IL-6, IL-10, MCP-1 and TNF protein levels in serum samples. Our studies demonstrate that exogenous TRX has anti-inflammatory properties and intracellular regulator activity in vivo and in vitro. These results have a therapeutic role in skin inflammation therapy.