L-Type Calcium Channel Blocker Enhances Cellular Delivery and Gene Silencing Potency of Cell-Penetrating Asymmetric siRNAs.

The efficient delivery of small interfering RNAs (siRNAs) to the target cells is critical for the pharmaceutical success of RNA interference (RNAi) drugs. One of the possible strategies to improve siRNA delivery is to identify auxiliary molecules that augment their cellular uptake. Herein, we performed a chemical library screening in an effort to discover small molecules that enhance the potency of cholesterol-conjugated, cell-penetrating asymmetric siRNAs (cp-asiRNAs). Interestingly, three compounds identified from the screen share a common dihydropyridine (DHP) core and function as L-type calcium channel blockers (CCBs). Using confocal microscopy and quantitative analysis of small RNAs, we demonstrated that the L-type CCBs increased the endocytic cellular uptake of cp-asiRNAs. Furthermore, these small molecules substantially improved the potency of cp-asiRNAs, not only in vitro but also in vivo on rat skin. Collectively, our study provides an alternative pharmacological approach for the identification of small molecules that potentiate the effects of therapeutic siRNAs.

Correlation between mast cell-mediated allergic inflammation and length of perfluorinated compounds.

Perfluorinated compounds (PFC) have widely been used in numerous applications including clothing, food packaging, and nonstick coating. With the widespread use of PFC, concerns regarding potential adverse health effects in humans and wildlife have increased. In spite of the known PFC-mediated immunotoxiciy, correlation with PFC and allergic inflammation still requires elucidation. The aim of this study was to examine the effect of four types of PFC (perfluoroheptanoic acid [PFHpA], perfluorononanoic acid [PFNA], perfluorodecanoic acid [PFDA], and perfluoroundecanoic acid [PFUnA]) on mast cell-mediated allergic inflammation in the presence of high-affinity immunoglobulin (Ig) E receptor (FcεRI) cross-linking. Among PFC family, long-chain PFDA and PFUnA increased release of histamine and ß-hexosaminidase by up-regulation of intracellular calcium levels in IgE-stimulated mast cells. In addition, PFDA and PFUnA enhanced gene expression of pro-inflammatory cytokines, such as tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, IL-6, and IL-8 by activation of nuclear factor-κB in IgE-stimulated mast cells. In ovalbumin (OVA)-induced model of systemic anaphylaxis in the presence of hypothermia, PFNA, PFDA, and PFUnA exacerbated allergic symptoms accompanied by elevation in serum histamine, TNF-α, IgE, and IgG1. Our data indicate that some PFC aggravated high-affinity IgE receptor (FcεRI)-mediated mast cell degranulation and allergic symptoms. Consequently, the results demonstrated that carbon-chain length of PFC may serve as a factor in allergic inflammation.

Association between perfluorooctanoic acid exposure and degranulation of mast cells in allergic inflammation.

Perfluorooctanoic acid (PFOA) has wide applications, including as a raw material for converted paper and packaging products. With the widespread use of PFOA, concerns regarding its potential environmental and health impacts have increased. In spite of the known hepatotoxicity and genotoxicity of PFOA, correlation with PFOA and allergic inflammation is not well known. In this study, the effect of PFOA on the degranulation of mast cells and mast cell-mediated allergic inflammation in the presence of FcεRI cross-linking was evaluated. In immunoglobulin (Ig) E-stimulated mast cells, PFOA increased the release of histamine and ß-hexosaminidase by the up-regulation of intracellular calcium levels. PFOA enhanced gene expression of several pro-inflammatory cytokines, including tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, IL-6, and IL-8 by the activation of nuclear factor (NF)-κB in IgE-stimulated mast cells. Also, PFOA exacerbated allergic symptoms via hypothermia, and an increase of serum histamine, TNF-α, IgE and IgG1 in the ovalbumin-induced systemic anaphylaxis. The present data indicate that PFOA aggravated FcɛRI-mediated mast cell degranulation and allergic symptoms. Copyright © 2016 John Wiley & Sons, Ltd.

Protective activity of kudzu (Pueraria thunbergiana) vine on chemically-induced hepatotoxicity: in vitro and in vivo studies.

BACKGROUND: Kudzu (Pueraria thunbergiana) root has long been used in Traditional Chinese Medicine. However, the vine of the kudzu plant has been considered waste material. This study aimed to investigate the hepatoprotective properties of the kudzu vine. METHODS: We created 0 %, 30 %, 70 %, and 95 % ethanolic kudzu vine extracts. The isoflavone contents of kudzu vine extract were quantified by high-performance liquid chromatography. Tertiary-butylhydroperoxide (t-BHP) was added to human liver-derived HepG2 cells, and the production of reactive oxygen species was measured in the presence and absence of kudzu vine extract. Antioxidant activity was evaluated in all kudzu vine extracts using a hydroxyradical scavenging assay. Thirty-five male Sprague-Dawley rats were divided into seven groups (n = 5); two groups were not given any extract or drug, one group was treated with 50 mg/kg silymarin orally for 5 days, and the remaining four groups were respectively treated with 100 mg/kg of 0%, 30%, 70%, or 95% ethanolic extract of kudzu vine orally once daily for 5 days. On day 5 the treatment groups and one untreated group were fed 0.75 ml/kg carbon tetrachloride (CCl4) to induce liver damage. Blood and liver tissue samples were collected 24 h after CCl4 administration for measurement of plasma alanine aminotransferase and aspartate aminotransferase, and concentration of malondialdehyde and glutathione in liver tissue. RESULTS: Puerarin was the most abundant isoflavone in kudzu vine extract. Kudzu vine extract significantly reduced the cytotoxicity and production of reactive oxygen species induced by t-BHP in a dose-dependent manner. Treatment with 0 % and 30 % ethanolic extracts of kudzu vine significantly lowered the plasma levels of alanine aminotransferase and aspartate aminotransferase in a CCl4-induced hepatotoxicity rat model (P < 0.05). Glutathione was significantly elevated in the 30 % ethanolic extract-treated group (P < 0.05), while the malondialdehyde level in liver tissue was significantly decreased in the 0 % and 30 % ethanolic extract-treated groups (P < 0.05). CONCLUSIONS: The kudzu vine is potentially highly beneficial in treating liver damage, as it scavenges reactive free radicals and boosts the endogenous antioxidant system.

Changes of oxidative/antioxidative parameters and DNA damage in firefighters wearing personal protective equipment during treadmill walking training.

[Purpose] The purpose of this study was to investigate the influence of personal protective equipment on the oxidant/antioxidant parameters and DNA damage in firefighters during training and recovery. [Subjects and Methods] Twelve male nonsmoking volunteer firefighters (35.1 ± 7.2 years) underwent two maximal treadmill training (9 METs, 6 km/h), within 2 weeks, one in regular clothes and one in personal protective equipment weighing 22.1 kg. Blood samples were obtained before, right after, and 40 min after training. Plasma conjugated dienes, total radical trapping antioxidant potential, erythrocytes antioxidant enzymes activities, and leukocyte DNA damage were measured. [Results] Wearing personal protective equipment during treadmill walking training resulted in increases of plasma conjugated dienes, total radical trapping antioxidant potential, and leukocyte DNA resistance to oxidative stress, which were recovered after in 40 min of rest. Erythrocyte antioxidant enzymes activities remained unchanged during the training either with regular clothes or personal protective equipment. [Conclusion] These results suggest that wearing personal protective equipment during firefighting work could induce oxidative stress, which was enough to produce DNA damage in leukocytes.

Artemisia capillaris inhibits atopic dermatitis-like skin lesions in Dermatophagoides farinae-sensitized Nc/Nga mice.

BACKGROUND: Artemisia capillaries Thunb. (AC) has been used to treat inflammatory and hepatic disorders such as hepatic injury, hepatic fibrosis and hepatitis. However, the efficacy of AC against atopic dermatitis (AD), an inflammatory disease, has not been examined. In the present study, AC was evaluated for anti-inflammatory and anti-AD effects using both in vitro and in vivo systems. METHODS: The contents of six compounds (chlorogenic acid, caffeic acid, isochlorogenic acid A, hyperoside, isoquercitrin and scoparone) in AC were simultaneously assayed using HPLC system. To evaluate the anti-inflammatory effect of AC, NO production was measured in RAW264.7 cell stimulated with 1 µg/mL LPS. Histamine levels were assayed in MC/9 cells stimulated with 50 nM PMA and 1 µM A23187. To examine the role of AC in vivo, AC (10 mg/mouse/day) was topically applied for four weeks the back and ears of Dermatophagoides farinae-sensitized Nc/Nga mice. Protopic ointment (0.1% tacrolimus) was used as a positive control. RESULTS: The contents of the six components in AC range from 0.44 to 43.14 mg/g. Chlorogenic acid (21.06 ± 0.08 mg/g) and isochlorogenic acid A (43.14 ± 0.12 mg/g) were major components in AC. AC inhibited NO and histamine production in cells respectively. In D. farinae-sensitized Nc/Nga mice, the topical application of AC reduced dermatitis scores, hemorrhage, hypertrophy and hyperkeratosis of the epidermis in the dorsal skin and ear. The treatment of AC also reduced the plasma levels of histamine (1.5 fold) and IgE (1.4 fold). CONCLUSIONS: Our results suggest that AC should be explored as a potential therapeutic agent to treat atopic dermatitis and analysis by HPLC will help to improve the quality of AC.

Angelicae Dahuricae Radix Inhibits Dust Mite Extract-Induced Atopic Dermatitis-Like Skin Lesions in NC/Nga Mice.

We examined whether Angelicae Dahuricae Radix (AR) suppresses the development of atopic dermatitis (AD)-like skin lesions induced by Dermatophagoides farinae in NC/Nga mice. To investigate the effect of AR, we measured the AD severity score, measured plasma levels of IgE and histamine, and performed histological analysis in NC/Nga mice. We also confirmed the anti-inflammatory effects of AR by measuring TARC/CCL17 production from LPS-treated RAW 264.7 cells and mRNA levels of TARC and MDC/CCL22 in TNF-α/IFN-γ-treated HaCaT cells. 10 mg/day of AR extract was applied for 4 weeks to NC/Nga mice. Both the AR extract and 0.1% tacrolimus suppressed the development of AD-like skin lesions and reduced dermatitis scores of the back and ear skin. AR extracts caused an inhibition of histological changes induced by repeated application of D. farinae and a reduction of IgE and histamine levels in plasma (P < 0.05). Furthermore, NO production in LPS-treated RAW 264.7 cells was diminished in a dose-dependent manner, and hTARC production and TARC and MDC mRNA levels in TNF-α/IFN-γ-treated HaCaT cells were diminished by AR. The inhibitory effect of AR on NO, TARC and MDC production may be associated with the suppression of AD-like skin lesions in D. farinae-induced NC/Nga mice.

Safety Evaluation of Yukmijihwang-tang: Assessment of Acute and Subchronic Toxicity in Rats.

Yukmijihwang-tang (YMJ; Liu wei di huang tang (China), Rokumigan (Japan)) has been used in the treatment of diseases including renal disorder, cognitive vitality, and diabetes mellitus. However, there is very little information regarding the toxicity of YMJ to give an assurance of safety for clinical treatment. To provide safety information for YMJ, we evaluated its acute and sub-chronic toxicity in rats. The single-dose toxicity of YMJ was examined using Sprague-Dawley rats. Rats were treated with YMJ extract orally at 0, 500, 1000, or 2000 mg/kg body weight. After a single administration, clinical signs were observed every day for two weeks, and body weights were measured five times, including an initial measurement on day 1 (the day of administration). In the sub-chronic oral toxicity study, YMJ was administered to rats at 0, 500, 1000, or 2000 mg/kg/day for 13 weeks. Mortalities, clinical signs, body weight changes, food and water consumption, ophthalmologic findings, urinalysis, hematological and biochemical parameters, gross findings, organ weights, and histological examination were monitored during the study period. We found no mortality and no abnormalities in clinical signs, body weights, and necropsy findings for any of the animals in the acute and sub-chronic studies following oral administration in the rat at up to 2000 mg/kg/day YMJ. YMJ may not have any single-dose toxicity; the LD(50) of YMJ was over 2000 mg/kg, and it is safe for rats. The no-observed-adverse-effect-level (NOAEL) was considered to be 2000 mg/kg/day.

Evaluation of genotoxicity of Yukmijihwang-tang, a herbal formula.

Yukmijihwang-tang (Liu wei di huang tang, Rokumigan; YMJ) has been used for body enrichment; however, little toxicological evaluation of YMJ has been performed to assure its safety for clinical treatment. To increase the safety information for YMJ, its genotoxicity was evaluated. There was no increase in the number of revertant colonies in four strains of Salmonella typhimurium or one strain of Escherichia coli at any concentration of YMJ studied, regardless of the including when dosed with YMJ metabolized with and S-9 microsomal fraction. YMJ significantly increased structural aberrations in Chinese hamster lung (CHL) cells at the high concentrations (2500 and 5000 µg/ml) in the presence or absence of metabolic activation by the S-9 microsomal fraction. Oral administration of YMJ at doses up to 2000 mg/kg did not increase the incidence of micronucleated polychromatic erythrocytes in bone marrow. These results suggest that YMJ is not genotoxic at the proper dose.

Inhibitory effects of heartwood extracts of Broussonetia kazinoki Sieb on the development of atopic dermatitis in NC/Nga mice.

We investigated the effects of a topically applied extract of the heartwood of Broussonetia kazinoki Sieb (B. kazinoki) on atopic dermatitis (AD)-like skin lesions induced by an extract of the house-dust mite Dermatophagoides farina in NC/Nga mice. We found that topically applied B. kazinoki extract suppressed the histological manifestations of AD-like skin lesions, and decreased the levels of plasma immunoglobulin E (IgE) and interleukin-4 (IL-4) in the mice. Moreover, B. kazinoki inhibited the induction of thymus-and-activation-regulated chemokine (TARC/CCL17), macrophage-derived chemokine (MDC/CCL22), and regulated-on-activation-normal T cell-expressed-and-secreted chemokine (RANTES/CCL5) in HaCaT cells activated by tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma). In conclusion, our results suggest that B. kazinoki extract has therapeutic advantages in the treatment of AD.

Identification of the Thioredoxin-Like 2 Autoantibody as a Specific Biomarker for Triple-Negative Breast Cancer.

Triple-negative breast cancer (TNBC) has a higher risk of death within 5 years of being diagnosed than the other forms of breast cancer. It is the second leading cause of death due to cancer among women. Currently, however, no diagnostic blood-based biomarker exists to identify the early stages of TNBC. To address this point, we utilized a human protein microarray system to identify serum autoantibodies that showed different expression patterns between TNBC and normal serum samples, and identified five autoantibodies showing TNBC-specific expression. Among them, we selected the thioredoxin-like 2 (TXNL2) autoantibody and evaluated its diagnostic relevance by dot blot analysis with the recombinant TXNL2 protein. We demonstrated that the TXNL2 autoantibody showed 2- to 6-fold higher expression in TNBC samples than in normal samples suggesting that serum TXNL2 autoantibodies are potential biomarkers for TNBC.

Endothelium-dependent induction of vasorelaxation by the butanol extract of Phellinus igniarius in isolated rat aorta.

The butanol extract of Phellinus igniarius (BPI) induced relaxation of the phenylephrin e-precontracted rat aorta in a dose-dependent manner, and its effect was abolished by the removal of functional endothelium. Pretreatment of the aortic tissues with N(G)-nitro-L-arginine methyl ester (L-NAME), methylene blue, or 1H-[1,2,4]-oxadiazole-[4,3-alpha]-quinoxalin1-one (ODQ) inhibited the vascular relaxation induced by BPI. BPI-induced vascular relaxations were also markedly attenuated by the addition of verapamil or diltiazem, while the relaxant effect of BPI was not blocked by pretreatment with indomethacine, glibenclamide, tetraethylammonium (TEA), atropine, or propranolol. Incubation of endothelium-intact rat aorta with BPI increased the production of cGMP in a dose-dependent manner. These results suggest that BPI dilates vascular smooth muscle via endothelium-dependent nitric oxide-cGMP signaling pathway, with the possible involvement of L-type Ca(2+) channels.

Vasodilatory and anti-inflammatory effects of the 1,2,3,4,6-penta-O-galloyl-beta-D-glucose (PGG) via a nitric oxide-cGMP pathway.

Vasorelaxant and anti-inflammatory effects of a 1,2,3,4,6-penta-O-galloyl-beta-d-glucose (PGG) isolated from the root barks of Paeonia suffruticosa and possible mechanisms responsible were investigated. PGG induced a concentration-dependent relaxation of the phenylephrine-precontracted rat aorta. This effect disappeared with the removal of functional endothelium. Pretreatment of the aortic tissues with either N(G)-nitro-L-arginine methyl ester (L-NAME) or 1H-[1,2,4]-oxadiazole-[4,3-alpha]-quinoxalin-1-one (ODQ) inhibited the relaxation induced by PGG. Incubation of human umbilical vein endothelial cells (HUVECs) or carotid arteries isolated from rats with PGG increased the production of cGMP in a dose-dependent manner, but this effect was blocked by pretreatment with L-NAME and ODQ, respectively. PGG treatment attenuated tumor necrosis factor-alpha (TNF-alpha)-induced nuclear factor-kappaB (NF-kappaB) p65 translocation in human umbilical vein endothelial cells. In addition, PGG suppressed the expression levels of adhesion molecules including intracellular cell adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) induced by TNF-alpha. TNF-alpha-induced monocyte chemoattractant protein-1 (MCP-1) expression was also attenuated by addition of PGG. PGG treatment inhibited cellular adhesion of U937 cells onto human umbilical vein endothelial cells induced by TNF-alpha. Taken together, the present study suggests that PGG dilates vascular smooth muscle and suppresses the vascular inflammatory process via endothelium-dependent nitric oxide (NO)/cGMP signaling.

The Leaves of Broussonetia kazinoki Siebold Inhibit Atopic Dermatitis-Like Response on Mite Allergen-Treated Nc/Nga Mice.

Broussonetia kazinoki Siebold. (B. kazinoki) has long been used in the manufacture of paper in Asian countries. Although B. kazinoki leaves (BK) have been employed in dermatological therapy, use of BK has not been tested in patients with atopic dermatitis (AD). Using Nc/Nga mice, which are genetically predisposed to develop AD-like skin lesions, we confirmed the efficacy of BK in AD treatment. BK extract was applied topically to Dermatophagoides farinae-induced AD-like lesions in Nc/Nga mice, and the effects were assessed both clinically and by measuring skin thickness on the back and ears. We measured the effects of BK extract on plasma levels of IgE and IL-4. We also measured the ability of BK extract to inhibit the secretion of hTARC in HaCaT cells after stimulation by TNF-α and IFN-γ. We found that BK extract significantly reduced ear and dorsal skin thickness and the clinical signs of AD, as well as significantly down-regulating the plasma levels of IgE and IL-4 (p<0.01 for each comparison). Moreover, 500 µg/mL of BK extract inhibited hTARC secretion in HaCaT cells by activated TNF-α/IFN-γ by about 87%. These findings suggest that topical application of BK extract has excellent potential in the treatment of AD.

Anti-aging Potential of Extracts Prepared from Fruits and Medicinal Herbs Cultivated in the Gyeongnam Area of Korea.

Many recent studies have focused on maintaining a healthy life by preventing and/or postponing the aging process. Numerous studies have reported that continuous exposure to reactive oxygen species can stimulate skin aging and that excessive accumulation of fat can cause an impaired skin barrier and tissue structure alterations. Thus, the maintenance of antioxidant homeostasis and the suppression of adipose accumulation are important strategies for skin anti-aging. Here, we prepared three types of extracts [whole juice, acetone-perchloric acid (PCA), and ethanol] from 20 fruits and medicinal herbs native to the Gyeongnam area of Korea. The total phenolic content of each extract was analyzed, and we observed higher total phenolic contents in the medicinal herbs. Consistent with this, the results of the oxygen radical absorbance activity capacity assay indicated that the in vitro antioxidant activities of the medicinal herb extracts were stronger than those of the fruit extracts. The fruits and medicinal herbs had strong effects on cell-based systems, including H2O2-induced oxidative stress in human keratinocytes and 3T3-L1 lipid accumulation. Nishimura Wase persimmon, Taishu persimmon, wrinkled giant hyssop, sweet wormwood, Chinese cedar, red perilla, tan shen, hiyodori-jogo, and cramp bark may be natural anti-aging materials with effective antioxidant and anti-adipogenic activities. Taken together, our findings may provide scientific evidence supporting the development of functional foods and nutraceuticals from fruits and medicinal herbs.

Antiatopic Dermatitis Effect of Artemisia iwayomogi in Dust Mice Extract-Sensitized Nc/Nga Mice.

Aims. Artemisia iwayomogi (AI) has been used for fever reduction, diuresis, and hepatoprotection in Korea. The present study was performed to evaluate the anti-inflammatory and antiatopic dermatitis effects of AI using both in vitro and in vivo systems. Methods. The compositions in AI were analyzed by HPLC. To determine the anti-inflammatory effects of AI, the production of nitric oxide (NO) was measured in lipopolysaccharide treated RAW264.7 cells. Histamine levels were assayed to evaluate the antiallergic effects on MC/9 cells stimulated with phorbol-12 myristate 13-acetate and A23187. Finally, AI (10 mg/mouse/day) was topically applied onto the backs and ears of Dermatophagoides farinae-sensitized Nc/Nga mice for four weeks. Results. Isochlorogenic acid A (20.63 ± 0.26 mg/g), chlorogenic acid (9.04 ± 0.08 mg/g), and scopoletin (8.23 ± 0.01 mg/g) were among the major components of AI. AI inhibited the NO and histamine productions in RAW264.7 and MC/9 cells, respectively. In the mice, the topical application of AI reduced the dermatitis scores in the dorsal skin and ears and reduced the plasma levels of IgE. Conclusions. These results suggest that AI might be explored as a potential therapeutic agent to treat AD, and that the analytic method using HPLC will facilitate the development of quality control for AI.

Vascular relaxation induced by aqueous extract of Lespedeza cuneata via the NO-cGMP pathway.

The aqueous extract of Lespedeza cuneata G. Don. (ALC) induced vasorelaxation of phenylephrine precontracted aorta in a dose-dependent manner. This effect disappeared in the absence of functional endothelium. Pretreatment of the aortic tissues with N(G)-nitro-L-arginine methyl ester (L-NAME), or 1H-[1,2,4]-oxadiazole-[4,3-α]-quinoxalin-1-one (ODQ) blocked ALC-induced vascular relaxation. Incubation of endothelium-intact thoracic aortic rings with ALC increased cGMP production. ALC-induced cGMP production was blocked by pretreatment with L-NAME or ODQ. ALC-induced vascular relaxation was also markedly attenuated by addition of verapamil or diltiazem, but was not blocked by pretreatment with indomethacine, glibenclamide, tetraethylammonium, atropine, or propranolol. The results suggest that ALC dilates vascular smooth muscle via endothelium-dependent NO-cGMP signaling.

Therapeutic effects of the oriental herbal medicine Sho-saiko-to on liver cirrhosis and carcinoma.

The traditional Chinese herbal medicine Sho-saiko-to is a mixture of seven herbal preparations that has long been used in the treatment of chronic liver disease. Various clinical trials have shown that Sho-saiko-to protects against the development of hepatocellular carcinoma in cirrhotic patients. However, the mechanism by which Sho-saiko-to protects hepatocytes against hepatic fibrosis and carcinoma is not yet known. Basic science studies have demonstrated that Sho-saiko-to reduces hepatocyte necrosis and enhances liver function. Sho-saiko-to significantly inhibits hepatic fibrosis by inhibiting the activation of stellate cells, the major producers of collagen in the liver, as well as by inhibiting hepatic lipid peroxidation, promoting matrix degradation, and suppressing extracellular matrix (ECM) accumulation. Furthermore, clinical trials have shown that Sho-saiko-to lowers the rate of hepatocellular carcinoma (HCC) development in patients with cirrhosis and increases the survival of patients with HCC. Unfortunately, some case reports have shown the side effects of Sho-saiko-to. Most of the side effects were interstitial pneumonia and acute respiratory failure induced by Sho-saiko-to in Japan. As a result of analyzing these case reports, the incidence and risk are increased by co-administration of interferon, duration of medication, and, high in an elderly population. This review discusses the properties of Sho-saiko-to with regards to the treatment of chronic liver diseases and suggests the side effects of Sho-saiko-to.

Genotoxicity assessment of a herbal formula, Ojeok-san.

ETHNOPHARMACOLOGICAL RELEVANCE: Ojeok-san (OJS, wuji powder, goshaku-san), a widely used herbal formula in traditional Korean medicine, is used to treat illnesses such as the common cold, fatigue and gastrointestinal disorders; however there is insufficient background information about its safety. To establish safety information for OJS, we evaluated its genotoxicity. MATERIALS AND METHODS: The ability of OJS to induce reverse mutations was evaluated in Salmonella typhimurium (TA100, TA1535, TA98 and TA1537) and Escherichia coli (WP2uvrA) in the presence or absence of the metabolic activation system (S-9 mix). Chromosomal aberrations were evaluated in response to OJS, and viability and metaphase were analyzed in Chinese hamster lung (CHL) cells in the presence or absence of S-9 mix. A micronucleus test was performed using bone marrow cells from male ICR mice. OJS was orally administered twice at a 24h interval at a dose of 500, 1000 and 2000 mg/kg in mice. RESULTS: There were no increases in the number of revertant colonies at any concentrations of OJS regardless of S-9 mix in all tester strains compared to the vehicle control. OJS did not significantly increase the number of structural aberration in CHL cells in the presence or absence of S-9 mix. The oral administration of OJS at doses up to 2000 mg/kg caused no significant increase in the number of micronucleated polychromatic erythrocytes (MNPCEs) and in the mean value for the ratio of PCE to total erythrocytes (PCE/(PCE+NCE)). NCE is normochromatic erythrocyte. OJS did not increase the incidence of MNPCEs in bone marrow. CONCLUSIONS: These results suggest that OJS is toxicologically safe on genotoxicity studies.

Simultaneous determination of liquiritin, hesperidin, and glycyrrhizin by HPLC-photodiode array detection and the anti-inflammatory effect of Pyungwi-san.

A high-performance liquid chromatographic method was developed and validated to determine liquiritin, hesperidin, and glycyrrhizin levels in a traditional Korean medicine, Pyungwi-san (PWS). Reverse-phase chromatography using a C18 column operating at 40oC, and photodiode array detection at 254 nm and 280 nm, were used for quantification of the three marker components of PWS. The mobile phase using gradient flow consisted of two solvent systems. Solvent A was 1.0% (v/v) aqueous acetic acid and solvent B was acetonitrile with 1.0% (v/v) acetic acid. Calibration curves were acquired with r (2) > 0.9999, and the relative standard deviation values (%) for intra- and inter-day precision were both less than 4.0%. The recovery of each compound was in the range 97.33-110.72%, with an relative standard deviation less than 6.0%. To provide information on the biological activity of PWS, anti-inflammatory action was evaluated. Production of nitric oxide and prostaglandin E(2) were measured using the Griess reagent and enzyme-linked immunosorbent assay, respectively. PWS showed inhibitory effect on prostaglandin E(2) production in LPS-treated RAW 264.7 cells.