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Objectives : Traditional medicine Oryeong-san (ORS) has been prescribed for a long time to treat light fever, thirst, dysuria, and accompanying edema. However, the acute toxicity and safety were not reported. In this study, we evaluated the potent acute toxicity and safety of ORS. Methods : ICR mice were used to evaluate acute toxicity and safety by oral administration of 0, 500, 1,000, 2,000mg/kg of ORS. Mortality, body weight, and clinical symptoms were observed, and organ weight and blood biochemical parameters were analyzed after necropsy. Results : We found no mortality and no toxic or abnormal clinical symptoms by administration of ORS. Comparing with control group, no significant alterations in organ weight and blood biochemical parameters were observed. Conclusions : ORS recognized as safe and non-toxic medicinal material, and median lethal dose considered to be over 2,000 mg/kg in both male and female ICR mice. Keyword : Oryeong-san (Wuling-san), Acute toxicity, Safety.

In order to promote the gastrointestinal (GI) track absorption of S-164, which is known to be efficacious in the treatment of thrombotic and liver protection. S-164 suspension, liposomal suspensions containing S-164, and cubosomal suspensions containing the same were prepared by a sonication method. The particles in the suspensions exhibited multi-modal distribution and the mean diameters were hundreds of nanometer. The zeta potentials of particles contained in the S-164 suspension, the liposomal suspension and the cubosomal suspension were all negative, ranging from −10 mV to −22 mV, and no significant agglomeration and sedimentation were observed for couples of weeks. The fluctuation in particle size with the pH change of medium and the time lapse was significantly damped by the liposome and the cubosome. On TEM photos, S-164 exhibited no structural feature, liposomal S-164 was spherical lamellar particles, and cubosomal S-164 exhibited stripes on their particles. Liposomes had little effect on the in v...

Objectives : Samchulgeonbi-tang (shenzhujianpi-tang) has been prescribed as one of traditional herbal medicine for treatment of stomach diseases since ancient time in Korea. Samchulgeonbi-tang extract was fermented by Lactobacillus spp. for improving the effect. However, the toxicity and safet y of fermented Samchulgeonbi-tang (FS) extract were not confirmed. Therefore, this study was performed to evaluate the acute toxicity and safety of FS extract.Methods : To evaluate the acute toxicity and safety of FS extract, several doses of FS extract, 0, 500, 1000 and 2000 ㎎/㎏, were orally administered to 20 male and 20 female ICR mice, respectively. After treatment with FS extract, we observed mortality, general toxicity, behavior and change of body weight for the 14 days. After 14 days of oral administration, all mice were sacrificed and hema tological parameters were analyzed from blood serum.Results : In present study, the toxic signs such as mortality or abnormal behaviors by FS extract were not observed. There are no significant differences between FS-treat ed group and control group in body weight, organ weights, and hematological parameters.Conclusions : The remarkable adverse effects by FS extract were not observed in ICR mice. Also, any death was not occurred at all treated FS doses, 500, 1000 and 2000 ㎎/㎏. T herefore, the approximate lethal dose (ALD) of FS extract may be more than 2000 ㎎/㎏.Key words : amchulgeonbi-tang (shenzhujianpi-tang), Fermented Samchulgeonbi-tang (FS), Acute toxicity, Safety, ALD

A Study on Acute Oral Toxicity of Pyungwi-san and Fermented Pyungwi-san in ICR MiceDoo-Rye Jang, Youn-Hwan Hwang, Ki-Youn JungJeong-Ho Ha, Hwa-Yong Park, Jin-Yeul MaKM-Based Herbal Drug Research Group, Korea Institute of Oriental MedicineObjectives: This study was conducted to investigate the acute toxicity of Pyungwi-san(Pingwei-san in Chinese) in ICR mice, according to K FDA and OECD guideline.Methods: In the present study, 15 male and female ICR mice administrate d singly by gavage at dose levels of 0 and 2000 mg/kg of Pyungwi-san. During the experimental period, no treatment-related death was observed. T here were no adverse effects on clinical signs, body weight, and gross findi ngs at all treatment groups.Results: These results showed that the single oral adminstration of Pyu ngwi-san (Pingwei-san) did not cause any toxic effect at the dose levels of 2000 mg/kg in rats.Conclusions: Taken together, the median lethal dose(LD 50 ) of Pyungwi-san (Pingwei-san) was considered to be over 2000 mg/kg body for both sexes.Key Words: Pyungwi-san, Acute toxicity, Mice, LD

KIOM-C suspension and cubosome/KIOM-C suspension were prepared in various buffer solution of different pHs by a sonication method. KIOM-C particles were unstable in terms of time-dependent change in the size and the zeta potential. The fluctuation in the size and the zeta potential of KIOM-C particles was effectively damped by cubosome. The shape of KIOM-C particles was irregular and the size was hundreds to thousands of nm. Cubosome/KIOM-C particles exhibited stripes on their surfaces and the size was hundreds of nm. Regardless of the pH values (pH 2.0–10.0) of media, baicalin, a major component of KIOM-C, was chemically stable for 48 hours at a room temperature. Cubosome markedly enhanced the stability of baicalin particles against sedimentation. EDTA significantly enhanced the in vitro small intestinal absorption of baicalin, but MO cubosome hardly promoted.

We evaluated the herb-drug interaction potential of Galgeun-tang (GGT) extracts, mediated by cytochrome P450 (CYP) inhibition/induction. Further, the effects of fermentation on the CYP-mediated herb-drug interaction potential of GGT extracts were determined. As measured by LC-ESI/MS/MS, GGT extracts (0-300μg/mL) showed no inhibitory activity toward eight CYP isoforms (1A2, 2A6, 2B6, 2C9, 2C19, 2D6, 2E1, and 3A4) in pooled human liver microsomes, suggesting that GGT may have low potential for herb-drug interactions mediated by CYP inhibition. Hepatic CYP expression and activity in rats treated with GGT extracts twice per day for 1week was examined. Among the tested CYP isoforms (1A1, 1A2, 1B1, 2B1, 2C11, 2E1, 3A1, 3A2, and 4A1), CYP1B1 and 4A1 were increased by GGT extracts. Hepatic activities of 7-ethoxyresorufin-O-deethylase, 7-pentoxyresorufin-O-depentylase, and chlorzoxazone 6-hydroxylase, but not midazolam hydroxylase were also elevated. These results raise the possibility that GGT extracts may increase the toxicity of environmental toxicants through the elevating CYP-dependent metabolic activation. Interestingly, the increases in CYP1B1 and CYP4A1 levels, and 7-ethoxyresorufin-O-deethylase, 7-pentoxyresorufin-O-depentylase, and chlorzoxazone 6-hydroxylase activities were attenuated by fermentation of GGT extract using Lactobacillus plantarum KFRI 402, but not 144. Further studies are needed to identify the CYP regulatory component(s) from GGT and determination its metabolism.

To identify the active compound arctigenin in Fructus Arctii (dried seed of medicinal plantArctium lappa) and to elucidate the inhibitory mechanism in melanogenesis, we analyzed melanin content and tyrosinase activity on B16BL6 murine melanoma and melan-A cell cultures. Water extracts of Fructus Arctii were shown to inhibit tyrosinase activity in vitro and melanin content inα-melanocyte stimulating hormone-stimulated cells to similar levels as the well-known kojic acid and arbutin, respectively. The active compound arctigenin of Fructus Arctii displayed little or no cytotoxicity at all concentrations examined and decreased the relative melanin content and tyrosinase activity in a dose-dependent manner. Melanogenic inhibitory activity was also identified in vivo with zebrafish embryo. To determine the mechanism of inhibition, the effects of arctigenin on tyrosinase gene expression and tyrosinase promoter activity were examined. Also in addition, in the signaling cascade, arctigenin dose dependently decreased the cAMP level and promoted the phosphorylation of extracellular signal-regulated kinase. This result suggests that arctigenin downregulates cAMP and the tyrosinase enzyme through its gene promoter and subsequently upregulates extracellular signal-regulated kinase activity by increasing phosphorylation in the melanogenesis signaling pathway, which leads to a lower melanin content.

In continuation of our research to find biological components from Panax stipuleanatus, four oleanane-type triterpenes (12 to 15) were isolated successively. Fifteen oleanane-type saponins (1 to 15) were evaluated for nuclear factor (NF)-κB activity using a luciferase reporter gene assay in HepG2 cells. Compounds 6 to 11 inhibited NF-κB, with IC50 values between 3.1 to 18.9 μM. The effects on inducible nitric oxide synthase and cyclooxygenase-2 by compounds 8, 10, and 11 were also examined using reverse transcription-polymerase chain reaction. Three compounds (8, 10, and 11) inhibited NF-κB activity by reducing the concentration of inflammatory factors in HepG2 cells.

Oyaksungisan (OY) is a traditional herbal formula broadly used to treat beriberi, vomiting, diarrhea, and circulatory disturbance in Asian countries from ancient times. The effect of OY on cancer, however, was not reported until now. In this study, we have demonstrated that OY inhibits cell proliferation and induces cell deathviamodulating the autophagy on human colon cancer cells. In HCT116 cells, OY increased the ratio of LC3-II/LC3-I, a marker of autophagy, and treatment with 3-MA, an inhibitor of autophagy, and considerably reduced the formation of autophagosomes. In addition, OY regulated mitogen-activated protein kinase (MAPK) cascades; especially, JNK activation was closely related with autophagy effect by OY in HCT116 cells. Our results indicate that autophagy induction is responsible for the antiproliferative effect by OY, despite the weak apoptosis induction in HCT116 cells. In conclusion, OY might have a potential to be developed as an herbal anticancer remedy.

Hwangryunhaedok-tang (HRT) has been long used as traditional medicine in Asia. However, inhibitory role of HRT is unclear in early stage of 3T3-L1 adipocyte differentiation related to signaling. In the present study, we investigated the inhibitory effects of HRT on upstream signaling of peroxisome proliferation-activity receptor-γ(PPAR-γ) and CCAAT/enhancer binding protein-β(C/EBP-β) expression in differentiation of 3T3-L1 preadipocytes. We found that HRT significantly inhibited the adipocyte differentiation by downregulating several adipocyte-specific transcription factors including PPAR-γ, C/EBP-α, and C/EBP-βin 3T3-L1 preadipocytes. Furthermore, we observed that HRT markedly inhibited the differentiation media-mediated phosphorylation of Raf/extracellular mitogen-activated protein kinase 1 (MEK1)/signal-regulated protein kinase 1/2 (ERK1/2) and phosphorylation of phosphoinositide-dependent kinase 1 (PDK1)/Akt. These results indicate that anti-adipogenesis mechanism involves the downregulation of the major transcription factors of adipogenesis including PPAR-γand C/EBP-αthrough inhibition of Raf/MEK1/ERK1/2 phosphorylation and PDK1/Akt phosphorylation by HRT. Furthermore, high performance liquid chromatography (HPLC) analysis showed HRT contains active antiobesity constituents such as palmatine, berberine, geniposide, baicalin, baicalein, and wogonin. Taken together, this study suggested that anti-adipogenesis effects of HRT were accounted by downregulation of Raf/MEK1/ERK1/2 pathway and PDK1/Akt pathway during 3T3-L1 adipocyte differentiation.

Obovatol has been reported biological activities such as muscle relaxative, anti-gastric ulcer, anti-allergic and anti-bacterial activities. The present study was undertaken to investigate the effect of diacetylated obovatol, an obovatol derivative, on rabbit platelet aggregation, and their possible molecular mechanisms. Effects of diacetylated obovatol on platelet activation including aggregation and serotonin secretion were examined. In addition, we investigated the effect of diacetylated obovatol on archidonic acid and metabolites liberation and intracellular calcium mobilization. Diacetylated obovatol concentration-dependently inhibited the washed rabbit platelet aggregation induced by collagen and arachidonic acid, suggesting that diacetylated obovatol may selectively inhibits collagen- and arachidonic acid-mediated signal transduction. In accordance with these results, diacetylated obovatol showed a concentration-dependent decrease in cytosolic Ca(2+) mobilization and serotonin secretion. However, diacetylated obovatol did not inhibit arachidonic acid liberation; on the other hand, diacetylated obovatol inhibited the formation of arachidonic acid metabolites such as thromboxane A(2), prostaglandin D(2) and 12-HETE through interfering with cyclooxygenase (COX)-1 and lipoxygenase (LOX) activities. The results demonstrated that diacetylated obovatol has antiplatelet activities through inhibition of COX-1 and LOX activities.

Obovatol has various biological activities, including anti-proliferative, neurotrophic, anti-fibrillogenic, anti-platelet, anti-fungal and anti-inflammatory activities. In this study, we investigated the effects of JJK694, a synthesized obovatol derivative, on rabbit platelet activation and its molecular mechanisms. JJK694 significantly inhibited washed rabbit platelet aggregation and serotonin secretion induced by collagen and arachidonic acid, but had little effect on thrombin- or U46619-induced aggregation. These results suggest that JJK694 selectively inhibits collagen- and arachidonic acid-mediated signaling. JJK694 also showed a concentration-dependent decrease in cytosolic Ca(2+) mobilization, but it had no effect on arachidonic acid liberation. On the other hand, it significantly inhibited the formation of arachidonic acid metabolites, including thromboxane A(2) (TXA(2)), prostaglandin D(2), and 12-hydroxy-5,8,10,14-eicosatetraenoic acid (12-HETE), by suppression of cyclooxygenase (COX)-1 and lipoxygenase (LOX) activities. These results indicate that JJK694 hasanti-platelet activities through inhibition of arachidonic acid metabolite production by suppression of COX-1 and LOX activities.

Ion oxide nanoparticles (IONPs) were prepared in monoolein (MO) cubic phases. Cubic phases were prepared by hydrating molten MO with Fe 2+ /Fe 3+ salt solutions of which concentrations were 0.0125 M/0.025 M, 0.00625 M/0.0125 M, and 0.003125 M/0.00625 M. When the concentration were greater than 0.0125 M/0.025 M, cubic phase were hardly formed possibly because MO can be readily oxidized at a high concentration of Fe 3+ . The Fe 2+ /Fe 3+ -loaded cubic phases were alkalized by adsorbing aqueous solutions of NaOH (0.1 M, 0.05 M, and 0.025 M) into the cubic phases for the co-precipitation of the ions within the cubic phases. The resulting IONPs were around 10 nm on TEM photos, and the concentration of ions had little effect on the sizes. And, the size was somewhat less than IONPs prepared in bulk phases, possibly because the co-precipitation of Fe 2+ /Fe 3+ will take place in the water channels of the cubic phases. The cubosomes containing IONPs were prepared by micronizing the cubic phases. Magnetic interaction between the cubosomes and a magnet were observed and the interaction was proportional to the amount of IONPs in the cubosomes.

Turmeric is a medicinal herb with various pharmacological activities. Curcumin, demethoxycurcumin(DMC) and bisdemethoxycurcumin(BDMC) were extracted from turmeric by dipping and ultrasonic wave method. And extraction efficiency was identified using solid phase extraction(SPE). The extracted sample were simultaneously separated and analyzed from three kinds of commercial C18 reversed-phase high performance liquid chromatography using water and acetonitrile as mobile phase with isocratic elution mode. Flow rate 1.0mL/min, injection volume 10µL and column oven temperature 40℃ at 425nm wavelength has been conducted. From the experimental results, the optimum mobile phase composition of water/acetonitrile 50/50vol% using RS tech column. It is evident that the amount of curcuminoids extracted(extraction time 4h) by 100% MeOH was higher than any aqueous MeOH composition. Finally, in 100% water extraction, the amount(mAU×mim) of curcuminoids extracted by SPE was 14.3 and 24.5 times respectively higher than ultrasonic wave and dipping method. The shown results can be applied as sources for pharmaceuticals and functional material.

For the simultaneous determination of ferulic acid, hesperidin, 6-gingerol and glycyrrhizin in Insampaedok-san, one of traditional oriental medicine prescription, a high performance liquid chromatography (HPLC) analytical method was developed. The separation was achieved on a SHISEIDO C18 column (S-5 μm, 4.6 mm I.D. × 250 mm). The mobile phase consisted of water with 0.1% trifluoroacetic acid and methanol in gradient elution at a flow rate of 1.0 mL/min. The UV wavelength was set at 250, 280 and 320 nm. The method was validated for linearity, precision and recovery. All calibration curves showed good linear regression (r2 > 0.9994) over the wide test ranges. LOD (limit of detection) and LOQ (limit of qualification) ranged from 0.02 to 0.14 μg/mL and 0.05 to 0.42 μg/mL, respectively. The relative standard deviation (RSD) of the intra- and inter-day test, precision test were within 2.26 and 2.45%, respectively, with accuracy from 95.95 to 108.22%. The recoveries of the four compounds were obtained between 94.30 and 106.34% with RSD less than 1.96%. The verified method was successfully applied to quality control of Insampaedok-san.

Soshiho-tang, one of the herbal prescriptions, has been used in treatment of chronic liver disease. In this study, the efficient high-performance liquid chromatography coupled with diode array detection and electrospray ionization tandem mass spectrometry (HPLC-DAD-ESI-MS) method was developed and validated for simultaneous determination of six marker compounds, namely homogentisic acid, baicalin, glycyrrhizin, saikosaponin A, 6-gingerol and ginsenoside Rg3 in Soshiho-tang. The analysis of six marker compounds was carried out using a C(18) column (SHISEIDO S-5 μm, 4.6 mm I.D. × 250 mm) and gradient elution with water - 0.1% trifluoroacetic acid and acetonitrile. The analytical method was validated for linearity, precision and accuracy. Calibration curve for six marker compounds showed good linear regression (r(2)0.9996). The limits of detection (LOD) and limits of quantification (LOQ) for analytes were ranged from 0.04 to 0.16 μg/mL and 0.12 to 0.47 μg/mL, respectively. The relative standard deviations (RSD%) of the precision test, intra- and inter-day, were less than 0.32% and 0.10%, respectively. All recoveries of the six marker compounds were ranged from 100.05% to 102.25% with RSD less than 0.63%. These results have shown that this developed method was efficient for the quality evaluation of Soshiho-tang.

Sochengryongtang (SCRT), a traditional herbal medicine, has been used for the treatment of cold for thousands of years. In this study, high performance liquid chromatography with diode array detector (HPLC-DAD) and electrospray ionization mass spectrometry (ESI-MS) was applied for the reliable and sensitive simultaneous analysis of 11 major components (homogentisic acid, ephedrine HCl, paeoniflorin, cinnamic acid, cinnamaldehyde, glycyrrhizin, 6-gingerol, schisandrin, methyl engenol, gomisin A, and gomisin N) in SCRT. Components were separated on a C18 column using gradient elution with 0.1% TFA water and acetonitrile and characterized based on the retention time in HPLC chromatogram, UV spectrum, and MS data. All calibration curves of major components proved to have good linearity (r2 > 0.9999). The LOQ and LOD ranged from 7.7 to 47.8 ng/mL and 23.5 to 131.2 ng/mL, respectively. This developed method showed good precision and accuracy with intra-day and inter-day (relative standard deviation, R.S.D.) val...

Monoolein (MO) cubosomes were investigated in terms of in vitro skin permeation enhancer of KIOM-MA-128 (MA-128), a natural product known to be efficacious against atopic dermatitis. First, an aqueous suspension of MA-128 was prepared by homogenizing the powder in Pluronic F-127 (a dispersant) solution in water. The Pluronic F-127 concentration and the pH have no significant effect on the size and the zeta potential of MA-128 particles. The mean diameters and the zeta potentials fell within 1000–1500 nm and −10 to −20 mV, respectively. The sedimentation rate of the particles was lower at a higher concentration of the polymeric dispersant, possibly because the polymeric surfactant can act as a spring and push away approaching particles. The size of MO cubosomes was tens to hundreds of nanometers and exhibited black and white stripes. Cumulative amount of MA-128 permeated through hairless mouse skin was obviously higher when the cubosome was included in the MA-128 suspensions. However, the cumulative permea...

In this study, the possible application of Lactobacillus strains as a functional starter culture to ferment Samso-Eum (SE), an oriental herbal medicine formula, and the production of bioactive isoflavones (daidzein, genistein) were investigated. Four strains of Lactobacillus (Lactobacillus plantarum KFRI 144, L. amylophilus KFRI 161, L. curvatus KFRI 166, and L. bulgaricus KFRI 344) were used for SE fermentation. Declines in pH and in viable cell counts during fermentation were investigated and the quantification of isoflavones using HPLC were performed after fermentation at 37°C for 48 h. All the tested Lactobacillus strains lowered the pH level to approximately 3.6 after 48 h and showed the highest level of growth at 24 h during SE fermentation. During the SE fermentation of the four Lactobacillus strains, the conversion of isoflavone glycosides (daidzin, genistin) into bioactive aglycones (daidzein, genistein) was observed in all of the fermentations, but with different rates depending on the strains. L. plantarum KFRI 144 and L. amylophilus KFRI 161 exhibited the highest bioconversion rate of isoflavone glycosides into their bioactive aglycones. These results demonstrate that L. plantarum KFRI 144 and L. amylophilus KFRI 161 have potentials as functional starter cultures for manufacturing fermented SE with higher isoflavone bioavailability.

Herb-drug interactions represent a serious problem as herbal medicine is used extensively in the modern world. This study investigated the effects of decursinol angelate on the pharmacokinetics of theophylline, a typical substrate of the cytochrome P450 1A2 enzyme, in rats. After 3 days of decursinol angelate pretreatment, on the fourth day, rats were administered decursinol angelate and theophylline concomitantly. Blood theophylline and its major metabolite [1-methylxanthine (1-MX), 3-methylxanthine (3-MX), 1-methyluric acid (1-MU), and 1,3-dimethyluric acid (1,3-DMU)] levels were monitored by liquid chromatography-tandem mass spectroscopy. The results indicated that theophylline clearance significantly decreased and the area under the concentration–time curve (AUC) increased in decursinol angelate (25 mg/kg)-pretreated rats administered theophylline (10 mg/kg). The elimination half-life (t1/2) of theophylline was increased by 20%. In the presence of decursinol angelate (25 mg/kg), the pharmacokinetic parameters of three metabolites (1-MX, 1,3-DMU, and 1-MU) were significantly altered (half-life for 1-MU, and AUC24 h for 1-MX, 1,3-DMU, and 1-MU). Our results suggest that patients receiving CYP1A2-metabolized drugs, such as caffeine and theophylline, should be advised of the potential herb-drug interaction to reduce the risk of therapeutic failure or increased toxicity of conventional drug therapy.

Background Malva verticillata seeds are used as a therapeutic medicine to treat kidney dysfunction in traditional Chinese medicine (TCM). TCM has suggested that herbal medicine tonifying kidney function may have beneficial effect on bone metabolism. Methods Osteoclastogenesis was examined in bone marrow macrophages by measuring tartrate-resistant acid phosphatase (TRAP) activity and counting the number of TRAP-stained multinuclear cells. The activation of receptor activator of nuclear factor-kB (RANK) ligand signaling, and the expression of c-Fos and nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1) were investigated by western blot analysis. Transcription factor and bone resorption marker mRNA levels were evaluated using real-time quantitative polymerase chain reaction. The bone resorption activity of mature osteoclast was examined in osteoclasts cultured on a hydroxyapatite-coated culture plate. Results A water extract of M. verticillata seeds (WEMV) inhibited osteoclastogenesis stimulated by RANKL. WEMV also strongly inhibited expression of c-Fos and NFATc1 as well as phosphorylation of c-Jun N-terminal kinase, p38, I-kBα, and phospholipase γ2. Furthermore, WEMV significantly attenuated osteoclast resorption activity and downregulated mRNA expression of resorption markers. Conclusion These results demonstrate that WEMV inhibits osteoclastogenesis and bone resorption by suppressing the RANKL signaling pathway and suggest that M. verticillata seeds may be used as a therapeutic candidate in complementary alternative medicine to treat pathological bone diseases.

Background Cortex Phellodendri (C. Phellodendri), the dried trunk bark of Phellodendron amurense Ruprecht, has been known as a traditional herbal medicine, showing several bioactivities. However, antiviral activity of C. Phellodendri aqueous extract (CP) not reported in detail, particularly aiming the prophylactic effectiveness. Methods In vitro CP antiviral activity evaluated against Influenza A virus (PR8), Vesicular Stomatitis Virus (VSV), Newcastle Disease Virus (NDV), Herpes Simplex Virus (HSV), Coxsackie Virus (H3-GFP) and Enterovirus-71 (EV-71) infection on immune (RAW264.7) and epithelial (HEK293T/HeLa) cells. Such antiviral effects were explained by the induction of antiviral state which was determined by phosphorylation of signal molecules, secretion of IFNs and cytokines, and cellular antiviral mRNA expression. Furthermore, Compounds present in the aqueous fractions confirmed by HPLC analysis and evaluated their anti-viral activities. Additionally, in vivo protective effect of CP against divergent influenza A subtypes was determined in a BALB/c mouse infection model. Results An effective dose of CP significantly reduced the virus replication both in immune and epithelial cells. Mechanically, CP induced mRNA expression of anti-viral genes and cytokine secretion in both RAW264.7 and HEK293T cells. Furthermore, the main compound identified was berberine, and shows promising antiviral properties similar to CP. Finally, BALB/c mice treated with CP displayed higher protection levels against lethal doses of highly pathogenic influenza A subtypes (H1N1, H5N2, H7N3 and H9N2). Conclusion CP including berberine play an immunomodulatory role with broad spectrum antiviral activity, due to induction of antiviral state via type I IFN stimulation mechanism. Consequently, C. Phellodendri could be a potential source for promising natural antivirals or to design other antiviral agents for animal and humans. Electronic supplementary material The online version of this article (doi:10.1186/s12906-016-1206-x) contains supplementary material, which is available to authorized users.

Background: Kiom-18 is a novel composition of Cinnamomum cassia, Pinus densiflora, Curcuma longa and Glycyrrhiza glabra. Curcuma longa and Glycyrrhiza glabra, which are traditional medicines in Asia, have been reported to demonstrate preventive effects against atherosclerosis; however, they have not yet been developed into functional atherosclerosis treatments. We therefore studied the anti-atherosclerotic effects and possible molecular mechanisms of Kiom-18 using vascular smooth muscle cells (VSMCs). Methods: To assess the anti-proliferative effect of Kiom-18 in vitro, we performed thymidine incorporation, cell cycle progression, immunoblotting and immunofluorescence assays in VSMCs stimulated by platelet derived-growth factor (PDGF)-BB. In addition, we used LDLr knockout mice to identify the effects of Kiom-18 as a preliminary result in an atherosclerosis animal model. Results: Kiom-18 inhibited platelet-derived growth factor (PDGF)-BB-stimulated-VSMC proliferation and DNA synthesis. Additionally, Kiom-18 arrested the cell cycle transition of G(0)/G(1) stimulated by PDGF-BB and its cell cycle-related proteins. Correspondingly, the level of p27(kip1) expression was upregulated in the presence of the Kiom-18 extract. Moreover, in an atherosclerosis animal model of LDLr knockout mice, Kiom-18 extract showed a preventive effect for the formation of atherosclerotic plaque and suppressed body weight, fat weight, food treatment efficiency, neutrophil count, and triglyceride level. Conclusions: These results indicate that Kiom-18 exerts anti-atherosclerotic effects by inhibiting VSMC proliferation via G(0)/G(1) arrest, which upregulates p27(Kip1) expression.

Skeletal muscle atrophy is a critical feature of cancer-induced cachexia, caused by pro-cachectic factors secreted by host cells and tumor cells. Therefore, blockade of these factors has considered a reasonable target for pharmacological and nutritional interventions to prevent skeletal muscle loss under cancer-induced cachexia. Citrus unshiu peel (CUP) has been used for treating the common cold, dyspepsia, and bronchial discomfort and reported to have pharmacological activities against inflammation, allergy, diabetes, and viral infection. In the present study, we observed that daily oral administration of water extract of CUP (WCUP) to male BALB/c mice bearing CT-26 adenocarcinoma remarkably reduced the losses in final body weight, carcass weight, gastrocnemius muscle, epididymal adipose tissue, and hemoglobin (Hb), compared with saline treatment. The levels of serum IL-6 and muscle-specific E3 ligases elevated by tumor burden were also considerably reduced by WCUP administration. In an in vitro experiment, WCUP efficiently suppressed the production of pro-cachectic cytokines in immune cells as well as cancer cells. In addition, WCUP treatment attenuated C2C12 skeletal muscle cell atrophy caused by cancer cells. These findings collectively suggest that WCUP is beneficial as a nutritional supplement for the management of cancer patients with severe weight loss.

Background KBH-1 is an herbal mixture of Saururus chinensis, Curcuma longa and Polygala tenuifolia. Each herb has been reported to have various pharmaceutical activities; however, the synergistic effect of this herbal composition on obesity has not yet been determined. We investigated the alleviation effect of KBH-1 and its possible molecular mechanism in obesity-induced hepatic steatosis and leptin resistance in the hypothalamus. Methods We used HepG2 cells, primary neuronal cells and a high-fat diet (HFD)-induced obesity rat model to determine the effect of KBH-1 in vitro and in vivo on hepatic steatosis and leptin resistance accompanied by obesity. To identify the alleviation effect on lipid accumulation, HepG2 cells stimulated by FFA were stained with Oil Red O; in addition, immunoblotting and qPCR were performed to determine the effect of KBH-1 on the activation of proteins and nuclear enzymes in HepG2 cells and the steatotic liver of HFD-induced obesity rats. To examine the effect of KBH-1 on the leptin resistance of the hypothalamus and its possible molecular mechanism, we examined the effect of KBH-1 on the activation of the leptin resistance-related protein in primary cultured cortical neuron cells and the hypothalamus of an HFD-induced obesity rat model. In addition, we used HPLC analysis to identify the standard compound of KBH-1. Results KBH-1 not only suppressed the lipid deposition in HepG2 cells exposed to free fatty acids (FFA) but also significantly down-regulated major factors in lipogenesis and up-regulated major factors in lipolysis. Similarly, in a HFD-induced obesity model, KBH-1 improved hepatic steatosis by alleviating the effects on lipogenic genes and kinases. In addition, KBH-1 significantly improved the leptin-mediated signals impaired by obesity or FFA in the obesity model and primary cultured cortical neuron cells. In addition, KBH-1 was analyzed to include six standard compounds using HPLC analysis, among these compounds, onji-saponin B and curcumin were potently suppressed the level of triglycerides. Conclusions KBH-1 exhibits alleviating effects by improving hepatic steatosis and leptin resistance by up-regulating the activation of AMPK and suppressing the expression of PPARγ. These findings show the potential of KBH-1 as a functional food supplement or preventive agent in the treatment of obesity. Electronic supplementary material The online version of this article (doi:10.1186/s12906-016-1265-z) contains supplementary material, which is available to authorized users.

Angelica gigas Nakai and its components are known to have neuroprotective, antiplatelet, and anticancer activities. The present study evaluated the in vitro and in vivo biopharmaceutical characterization of Angelica gigas component substances, including decursin (the main substance), decursinol angelate (decursin isomer), JH714 (ether form of decursin) and epoxide decursin (epoxide form of decursin). Decursin, decursinol angelate and JH714 exhibited acceptable metabolic stability (50%) in liver microsomes from human and higher bound fraction (90%) in human plasma operating ultrafiltration. Decursin and decursinol angelate in CYP1A2 and CYP2C19 indicated less than 50% CYP activity, suggesting inhibition of the CYP isoforms using Vivid® CYP screening kit. JH714 only showed an apparent permeability coefficient of10 × 10⁻⁶ cm/s in MDCK cells, suggesting that it is poorly absorbed. Blood brain barrier permeability was examined after oral administration to male Sprague-Dawley (SD) rats, and pharmacokinetic studies were performed after oral and intravenous administration of 10 mg/kg compounds. Decursin, decursinol angelate and JH714 showed ratios of compound concentration in brain with respect to plasma (Cbrain/Cplasma) of1.5, suggesting good brain/plasma ratio at 0.5, 1, 3, and 5 h. In contrast, Cbrain/Cplasma was0.5 for epoxide decursin. For all test compounds,1.5% of the dose remained in GI tract after 8 h, and the excretion rate in urine was0.5% which suggests that gastro intestinal tract may be major site of disposition following oral administration. Finally, these results may be useful for the design of dosage regimens of decursin and its derivatives.

The influenza virus is one of the important respiratory risks affecting humans which require effective treatments. Some oriental medicines are currently treated for common cold as well as influenza infection. Previous studies have reported that the therapeutic properties of MA-128 are able to treat psoriasis antiasthmatic and atopic dermatitis. In this study, we investigated the therapeutic properties of the novel herbal medicine, MA-128, for the treatment of influenza virus infection by oral administration. MA-128 is an active natural biological compound from herbal-marine origin. The results showed that oral administration of mice with MA-128 could confer a survival benefit against Type A influenza virus infection. Daily oral administration of MA-128 delayed the death in infected mice for three days against mouse adapted H3N2 (A/Philippines/2/82). However, it protected more than 60% of mice from lethal infection of 2009 pandemic H1N1 (A/Korea/CJ01/2009) influenza virus. In addition, lung viral titers weresignificantly reduced at 7 days of post infection (~100 times) compared with mock-treated mice and viruses were cleared at 9 dpi only in MA-128 treated groups. This study demonstrated that the novel herbal medicine, MA-128, have the potentials as a herbal remedy against influenza A viruses.

Ethnopharmacological relevance Samul-tang (Si-Wu-tang in Chinese, Shimotsu-to in Japanese), widely used in eastern Asia, is composed of Angelica gigas (Angelicae Gigantis Radix), Cnidium officinale (Cnidii Rhizoma), Paeonia lactiflora (Paeonia Radix) and Rehmannia glutinosa (Rehmanniae Radix Preparata). Paeoniflorin, one of active components in Samul-tang has anti-platelet, anti-inflammation, anti-cancer and neuroprotective properties. However, there is no information about the effects of gender and food intake on the pharmacokinetics of paeoniflorin till now. Aim of the study This study was conducted to investigate whether food and gender could influence pharmacokinetic profiles of paeoniflorin after oral administration of Samul-tang. Materials and methods Male and female rats were administered with a single oral dose of Samul-tang equivalent to 80 mg/kg of paeoniflorin. Plasma concentrations of paeoniflorin were measured by high-performance liquid chromatography. The statistical differences of each group were evaluated using the analysis of variance (ANOVA) or Student t-test. Results The pharmacokinetic parameters of paeoniflorin were not significant different by gender difference. However, the maximum plasma concentration (Cmax, 0.47±0.29 μg/mL versus 1.10±0.35 μg/mL), area under the concentration-time curve (AUC0→∞, 1.41±0.89 h·μg/mL versus 3.12±1.61 h·μg/mL) and relative bioavailability (Frel=2.21) of fed rats were significantly increased in comparison with those of fasted rats (P Conclusion Taken together, food intake can affect both the rate and extent of absorption of paeoniflorin when Samul-tang was administered orally. Furthermore, this study demonstrates a readily preparative HPLC method in the research of traditional herbal medicine.

Objectives : This study was carried out to investigate the acute toxicity and safety of -(-) and fermented -(-) extract. Methods : To evaluate their acute toxicity and safety, 0(control group), 1250, 2500 and 5000 mg/kg of -(-) and fermented -(-) extracts were orally administered to 35 male and 35 female ICR mice. After a single administration, we observed survival rates, general toxicity, changes of body weight, and autopsy. Results : Compared with the control group, we could not find any toxic alteration in any of treated groups (1250, 2500 and 5000 mg/kg). Conclusions : of -(-) and fermented -(-) were over 5000 mg/kg and it is very safe to ICR mice.

Three major curcuminoids in turmeric (Curcumalonga), curcumin, demethoxycurcumin, and bisdemethoxycurcumin,were efficiently extracted by optimizing extraction condition andsimultaneously analyzed and identified using reverse phase high-performance liquid chromatography, thin-layer chromatography,and liquid chromatography-mass spectrometry method. Thehighest yield of extraction amount 0.279 g, 9.30% was obtainedby dipping method with extraction time of 7 h.Keywords curcuminoids · reverse phase high-performance liquid chromatography · simultaneous analysis ·thin-layer chromatography · turmeric국내·외 연구자들은 과학적인 방법으로 효율적인 삶의 질을찾기 위해 육상 및 해양 천연물로부터 유용성분들을 얻기 위한연구를 오래전부터 시도하고 있다. 특히 천연보존료 개발의 목적으로 허브류와 한약재 연구가 활발히 이루어지고 있으며(Naik등, 2011), 이 중에서 식품 및 한약재 소재로 잘 알려진 강황은오래전부터 아시아와 동남아에서 재배되어온 향신료의 성분이다(Choi, 2009; Oh 등, 2011). 효능 및 효과를 살펴보면 천연색소들은 오랫동안 항산화, 항염증, 해독작용 등의 다양한 생리활성이 있는 것으로 알려져 있어 그 기능적 가치가 더욱 높아지고 있다(Singh 등, 2010; Zhao 등, 2011). 또한, 황색 색소로서 curcumin이 0.3% 들어있고 특유한 향이 있는 정유로써turmerone과 dehydroturmerone이 1~5% 들어 있다(Choi, 2009).특히 curcuminoids의 curcumin (MW=368), demethoxycurcumin(DMC MW=338), bisdemethoxycurcumin (BDMC MW=308)은 주된 생리활성물질로 보고되었다(Li 등, 2011). 이러한 생리활성물질을 얻기 위한 방법으로 전형적인 용매추출법이 주로 사용되고 있으며(Wakte 등, 2011), 분리기술의 중요성은 더 주목받고 있다. 따라서 제한된 기술과 맞춤형 고순도, 중순도, 저순도의 기술적 조절이 가능해야 할 것이다(Jin 등, 2006). 이에 생물분리기술은 혼합물을 신속하고 고순도로 정제하는 기술뿐만아니라 생산을 위한 제조용 공정에도 많이 사용되고 탁월한 분리 능을 갖고 있다(Row, 1995). 2000년대 국내 바이오산업의네트워크 특성은 한방 및 바이오 산업체 원료 및 제품 품질검사, 제품개발(제제개발, 시범생산) 활성화, 천연물소재 정보제공및 효능탐색, 기업과의 공동 연구개발을 통한 천연물 연구분야가 활발히 이루어지고 있다. 따라서 기능성 식품, 의약품 및 기능성소재 개발을 위한 관능적으로 우수한 표준화된 추출법 개발과 생리적 특성 비교를 통해 대량생산을 위한 벨리데이션의원천기술확보가 필요하다(Jung 등, 2004).본 연구에서는 이러한 고부가가치를 얻기 위한 정량 및 정성분석방법으로 Liquid Chromatography-Mass Spectrometry,HPLC 및 Thin-Layer Chromatography (TLC)를 쉽게 접근하여빠르게 분석 및 정제하고(Zhang 등, 2008), curcumin, DMC및 BDMC를 추출하여 추출량의 최적조건을 제시하였으며, 효

In this study, an effective high performance liquid chromatography–diode array detector (HPLC–DAD) method was established for simultaneous determination of six marker compounds, ephedrine hydrochloride, 6-gingerol, glycyrrhizin, hesperidin, imperatorin and ferulic acid, in a Korean traditional prescription, Oyaksungisan, which is used for hemiplegia, arthralgia and paralysis. The six marker compounds of Oyaksungisan were separated on a LUNA C18 column (S-5 μm, 4.6 mm I.D. × 250 mm) at a column temperature of 35°C. The gradient elution was composed of water with 0.1% trifluoroacetic acid and methanol. The detection UV wavelengths were set at 207, 250, 280 and 320 nm. Calibration curves for the six compounds showed good linear regressions (R 2 > 0.9999). The limits of detection and limits of quantification were within the ranges 0.003–0.01 and 0.01–0.04 μg/ml, respectively. The relative standard deviation (RSD) values of intra- and inter-day testing were within the ranges 0.10–1.82 and 0.04–1.59%, respectively. The results of the recovery test were 95.05–104.27% with a RSD value of 0.11–1.85%. In conclusion, the simultaneous determination method developed was useful in the quality evaluation of Oyaksungisan.

Ssanghwa-tang is a medicinal formula that is widely prescribed in Korea to decrease fatigue after an illness. Fermented herbal medicines might be made more efficacious than conventional herbal medicines by increasing the absorption and bioavailability of the active compounds. In this study, Ssanghwa-tang was fermented to produce bioconversion compositions using Lactobacillus fermentum, and six peaks were decreased, four peaks were increased and one peak newly appeared in the HPLC-DAD chromatogram. The structures of the newly-appearing compound (1) and increased (2–5) compounds were identified as follows using NMR and MS: liquiritigenin (1), nodakenetin (2), cinnamyl alcohol (3), decursinol (4), and benzoic acid (5). The decreased compounds were identified to be paeoniflorin (6), liquiritin (7), nodakenin (8), cinnamaldehyde (9), decursin (10), and decursinol angelate (11) using HPLC-DAD analysis with authentic compounds. The high performance liquid chromatography method was used to quantify the eleven constituents in Ssanghwa-tang and fermented Ssanghwa-tang. All calibration curves of the standard compounds exhibited excellent linearity with a R2 > 0.9940.

Objectives：The purpose of this study was to investigate the changes in the contents of constituents in Samultang and its fermentations with 10 species of lactic acid bacteria.Methods：Ten strains of lactic acid bacteria, Lactobacillus casei, L. acidophilus , L. casei, L. plantarum, L. amylophilus, L. curvatus, L. delbruekil subsp. lactis, L. casei , B. breve, and B. thermophilum, were used for the fermentation of Samultang. The increased and decreased constituents were identified using HPLC/DAD and various liquid chromatographic techniques, and the structure was elucidated using NMR and MS. These compounds were quantitatively analyzed using an HPLC/DAD system.Results：A remarkably increased component was identified to be nodakenet in and a decreased component was determined to be nodakenin. The fermentation of the ten lactic acid bacteria demonstrated that the decomposable rate of these two compounds in fermented Samultang were different. Samultang fermented by L. plantarum showed the most remarkable changes.Conclusion：Nodakenetin was identified as bioconversion component after fer mentation and L. plantarum was discovered the best bacteria to increase the component.Key words：Samultang, fermentation, herbal medicine, bioconversion, nodake netin, nodakenin.

This study was conducted to determine if oral administration of the novel herbal medicine, KIOM-MA, and itsLactobacillus acidophilus-fermented product, KIOM-MA128, has therapeutic properties for the treatment of atopic dermatitis (AD). Using AD-induced BALB/c mice by Ovalbumin and aluminum hydroxide, the effectiveness of KIOM-MA and KIOM-MA128 on AD was evaluated. Oral administration of KIOM-MA and KIOM-MA128 reduced major clinical signs of AD including erythema/darkening, edema/papulation, excoriations, lichenification/prurigo, and dryness. Interestingly, KIOM-MA128 more significantly improved AD-related symptoms including decrease of IgE level in the plasma as well as reduction of scratching behavior, skin severity in the AD BALB/c model. HPLC analysis showed the significant changes in the constituent patterns between KIOM-MA and KIOM-MA128. Our results suggest that both KIOM-MA and KIOM-MA128 have potential for therapeutic reagent for the treatment of AD, and further, the efficacy is significantly enhanced byL. acidophilusfermentation via increases in its indicator molecule.

Sipjeondaebotang (SJ) has been used as a traditional drug in east-Asian countries. In this study, to provide insight into the biological effects of SJ and SJ fermented by Lactobacillus, we investigated their effects on lipopolysaccharide (LPS)-mediated inflammation in macrophages. The investigation was focused on whether SJ and fermented SJ could inhibit the production of pro-inflammatory mediators such as prostaglandin (PG) E2and nitric oxide (NO) as well as the expressions of cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS), tumor necrosis factor (TNF)-α, mitogen-activated protein kinases (MAPKs) and nuclear factor (NF)-κB in LPS-stimulated RAW 264.7 cells. We found that SJ modestly inhibited LPS-induced PGE2, NO and TNF-α production as well as the expressions of COX-2 and iNOS. Interestingly, fermentation significantly increased its inhibitory effect on the expression of all pro-inflammatory mediators. Furthermore, fermented SJ exhibited increased inhibition of p38 MAPK and c-Jun NH2-terminal kinase (JNK) MAPK phosphorylation as well as NF-κB p65 translocation by reduced IκBα degradation compared with either untreated controls or unfermented SJ. High performance liquid chromatography (HPLC) analysis showed fermentation by Lactobacillus increases liquiritigenin and cinnamyl alcohol contained in SJ, which are known for their anti-inflammatory activities. Finally, SJ fermented by Lactobacillus exerted potent anti-inflammatory activity by inhibiting MAPK and NF-κB signaling in RAW 264.7 cells.

Hwangryun-haedok-tang (HRT) is the common recipe in traditional Asian medicine, and microbial fermentation is used for the conventional methods for processing traditional medicine. We investigated the inhibitory effect of then-butanol fraction of HRT (HRT-BU) and fHRT (fHRT-BU) on the RANKL-induced osteoclastogenesis in bone-marrow-derived macrophages. mRNA expression of osteoclastogenesis-related genes were evaluated by real-time QPCR. The activation of signaling pathways was determined by western blot analysis. The marker compounds of HRT-BU and fHRT-BU were analyzed by HPLC. The inhibitory effect of HRT or fHRT on ovariectomy-induced bone loss were evaluated using OVX rats with orally administered HRT, fHRT (300, 1000 mg/kg), or its vehicle for 12 weeks. fHRT-BU significantly inhibited RANKL-induced osteoclastogenesis, and phosphorylation of p38, IKKα/β, and NF-κBp65 compared to HRT-BU. In addition, fHRT-BU also significantly inhibited the mRNA expression of Nfκb2, TNF-α, NFATc1, TRAP, ATPv0d2, and cathepsin K. Furthermore, administration of fHRT had a greater effect on the increase of BMD, and greater improved bone microstructure of the femora than that of HRT in ovariectomy rats. This study demonstrated that bacterial fermentation enhances the inhibitory effect of HRT on osteoclastogenesis and bone loss. These results suggest that fermented HRT might have the beneficial effects on bone disease by inhibiting osteoclastogenesis.

KIOM-MA was recently reported as a novel herbal medicine effective for atopic dermatitis and asthma. In this study, we have demonstrated the inhibitory effect of KIOM-MA on proinflammatory mediator produced in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. KIOM-MA significantly inhibited the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) as well as nitric oxide (NO) and prostaglandin E2(PGE2). Consistent with the inhibitory effect on PGE2, KIOM-MA suppresses the LPS-induced migration of macrophages and gelatinase activity and the expression of matrix metalloprotease-9 (MMP-9) in a dose-dependent manner. Additionally, KIOM-MA showed a strong suppressive effect on the inflammatory cytokines production such as tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6). We also found that KIOM-MA inhibits the activation of nuclear factor-κB (NF-κB) and represses the activity of extracellular signal-regulated kinase (ERK), p38, and c-Jun NH2-terminal kinase (JNK) mitogen-activated protein kinases (MAPKs). Taken together, we elucidated the mechanism of anti-inflammatory effect of KIOM-MA using RAW 264.7 cells stimulated by LPS.

This study was conducted to determine if oral administration of the novel herbal medicine, MA, and itsLactobacillus acidophilusfermented product, MA128, have therapeutic properties for the treatment of asthma. Asthma was induced in BALB/c mice by systemic sensitization to ovalbumin (OVA) followed by intratracheal, intraperitoneal, and aerosol allergen challenges. MA and MA128 were orally administered 6 times a week for 4 weeks. At 1 day after the last ovalbumin exposure, airway hyperresponsiveness was assessed and samples of bronchoalveolar lavage fluid, lung cells, and serum were collected for further analysis. We investigated the effect of MA and MA128 on airway hyperresponsiveness, pulmonary eosinophilic infiltration, various immune cell phenotypes, Th2 cytokine production, OVA-specific IgE production, and Th1/Th2 cytokine production in this mouse model of asthma. In BALB/c mice, we found that MA and MA128 treatment suppressed eosinophil infiltration into airways and blood, allergic airway inflammation and AHR by suppressing the production of IL-5, IL-13, IL-17, Eotaxin, and OVA-specific IgE, by upregulating the production of OVA-specific Th1 cytokine (IFN-γ), and by downregulating OVA-specific Th2 cytokine (IL-4) in the culture supernatant of spleen cells. The effectiveness of MA was increased by fermentation withLactobacillus acidophilus.

Hwangryunhaedok-Tang (HR) and berberine-containing single herbs are used to treat bacterial infection and inflammatory diseases in eastern Asia. The combination of berberine-containing herbal medicines and ciprofloxacin can be an excellent antibacterial chemotherapy against multidrug resistance bacteria. To evaluate the pretreatment effect of berberine and HR, vehicle, berberine (25 and 50 mg/kg/day), and HR (1.4 g/kg/day) were daily administered to rats for five consecutive days. On day 6, ciprofloxacin was administered (10 mg/kg, i.v. and 20 mg/kg, p.o.) to rats. To assess cotreatment effect of berberine and ciprofloxacin, berberine (50 mg/kg) and ciprofloxacin (20 mg/kg) were coadministered by single oral gavage. Pharmacokinetic data were estimated by noncompartmental model. Compared with ciprofloxacin alone (control group), coadministration of berberine (50 mg/kg) and ciprofloxacin significantly decreasedCmaxof ciprofloxacin (P<0.05). In addition, the pretreatment of berberine (50 mg/kg/day) and HR (1.4 g/kg/day) significantly decreasedCmaxandAUC0→∞, compared with control group (P<0.05). The oral bioavailability of ciprofloxacin was reduced by cotreatment of berberine and pretreatment of berberine and HR. Our results suggest that the expression of P-glycoprotein and organic anion and/or organic cation transporters (OAT/OCT) could take a role in reduced oral bioavailability of ciprofloxacin by berberine and HR.

Gangliosides play an important role in neuronal differentiation processes. The regulation of ganglioside levels is related to the induction of neuronal cell differentiation. In this study, the ST8Sia5 gene was transfected into mESCs and then differentiated into neuronal cells. Interestingly, ST8Sia5 gene transfected mESCs expressed GQ1b by HPTLC and immunofluorescence analysis. To investigate the effects of GQ1b over-expression in neurogenesis, neuronal cells were differentiated from GQ1b expressing mESCs in the presence of retinoic acid. In GQ1b expressing mESCs, increased EBs formation was observed. After 4 days, EBs were co-localized with GQ1b and nestin, and GFAP. Moreover, GQ1b co-localized with MAP-2 expressing cells in GQ1b expressing mESCs in 7-day-old EBs. Furthermore, GQ1b expressing mESCs increased the ERK1/2 MAP kinase pathway. These results suggest that the ST8Sia5 gene increases ganglioside GQ1b and improves neuronal differentiation via the ERK1/2 MAP kinase pathway.

Herbal Medicine Improvement Research Center, Korea Institute of Oriental Medicine, Daejeon 305-811, Korea.ABSTRACT : Galgeun-tang (GGT) is a traditional medicinal formula that is widely prescribed to treat cold, asthma, andhives in Korea. Fermented herbal medicines can be made more effective than normal herbal medicines by increasing theabsorption and bioavailability of the active compounds. In this study, we fermented Galgeun-tang to produce bioconversionconstituents using Lactobacillus plantarum (GGT144), and found that four peaks were decreased, three peaks wereincreased and two new peaks appeared in the HPLC-DAD chromatogram. After HPLC-DAD-guided fractionation of thenewly-appearing compounds (1 and 5) and the increased (6, 7, and 9) compounds, the structure of the compounds was deter-mined using NMR and MS. Using this approach the compounds were identified to be pyrogallol (1), daidzein (5), liquiritige-nin (6), cinnamyl alcohol (7), and formononetin (9), respectively. In addition, the decreased compounds were identified to bedaidzin (2), liquiritin (3), ononin (4), and cinnam aldehyde (8) using HPLC-DAD analysis with standard compounds. Thehigh performance liquid chromatography method was used to quantify the nine constituents in GGT and GGT144. All cali-bration curves of the standard compounds displayed excellent linearity with a R

Aim of the study The root of Angelica gigas Nakai (Korean “Dang-Gui”) has been used to treat female afflictions and anemia in traditional oriental herbal medicine since ancient times in Korea. The objective of this study is to identify the anti-cancer mechanism induced by A. gigas extract including decursin and decursinol angelate in human cervical cancer cells for scientific evidence regarding herbal therapy. Materials and methods The extract of A. gigas , decursin, and decursinol angelate was used to evaluate the anti-cancer effects on HeLa cells. Cell viability was evaluated by MTT assay and cell cycle regulation was analysed by FACS program. The expression of apoptotic proteins was confirmed by Western blot analysis and proteome profiler array. Results The extract of A. gigas containing decursin and decursinol angelate exhibited anti-cancer effect on HeLa cells through the significant activation of caspases and the cleavage of PARP. Expression of TRAIL and TRAIL receptors was also increased by two active components of A. gigas , respectively. In particular, two components regulated the expression of apoptosis-related proteins such as Bcl-2, Bcl-x, survivin, cIAP-1, -2, XIAP, etc. Conclusions These results suggest that TRAIL expression induced by extract of A. gigas stimulates the extrinsic and intrinsic apoptosis pathway by activating caspase-8 and caspase-9, respectively. Thus, extract of A. gigas including decursin and decursinol angelate takes a role in inhibition of cell-proliferation and activation of apoptosis in cervical cancer and could be developed as therapeutic anti-cancer agent against cervical cancer.

A Gram-positive, endospore-forming, new Bacillus species, strain BL4-6(T), was isolated from tidal flat sediment of the Yellow Sea. Strain BL4-6(T) is a straight rod, with motility by peritrichate flagella. The cell wall contains meso-diaminopimelic acid, and the major respiratory quinone is menaquinone-7. The major fatty acids are iso-C(15:0) and summed feature 3 (containing C(16:1) ω7c/iso-C(15:0) 2OH, and/or iso-C(15:0) 2OH/C(16:1) ω7c). Cells are catalase-positive and oxidase-negative. The G+C content of the genomic DNA is 38.0 mol%. Based on a comparative 16S rRNA gene sequence analysis, the isolate belongs to the genus Bacillus, forms a clade with the Bacillus cereus group, and is closely related to Bacillus mycoides (98.5%), Bacillus cereus (98.5%), Bacillus anthracis (98.4%), Bacillus thuringiensis (98.4%), Bacillus weihenstephanensis (98.1%), and Bacillus pseudomycoides (97.5%). The isolate showed less than 85% similarity of the gyrA gene sequence and below 95% similarity of the rpoB gene sequence to the members of this group. DNA-DNA relatedness between strain BL4-6(T) and B. cereus group was found to be in a range of 22.8-42.3%, and thus BL4-6(T) represents a unique species. On the basis of these studies, strain BL4-6(T) (=KCTC 13319(T) =JCM 15802(T)) is proposed to represent the type strain of a novel species, Bacillus manliponensis sp. nov.

Yukmijihwang-tang (YMT) is a traditional herbal medicine known to enhance memory in brain injury models. The aims of this study were to evaluate the inhibitory effect of YMT on osteoclast differentiation and to determine its molecular mechanism of action. YMT dose-dependently inhibited receptor activator for nuclear factor-κB (NF-κB) ligand (RANKL)-induced tartrate-resistant acid phosphatase (TRAP) activity and the formation of multinucleated osteoclasts in RAW264.7 cells. In addition, quantitative reverse transcription-polymerase chain reaction showed that YMT significantly decreased RANKL-induced expression of osteoclast differentiation-specific genes (TRAP, matrix metalloproteinase-9, cathepsin K, and the d2 isoform of vacuolar ATPase V(0) domain). Furthermore, YMT inhibited RANKL-induced phosphorylation of mitogen-activated protein kinases (extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38), phosphorylation of I-κBα, phosphorylation of NF-κB p65, and the expression of transcription factors Fra-2 and nuclear factor of activated T-cells, cytoplasmic 1. Furthermore, YMT inhibited the bone-resorptive activity of differentiated osteoclasts, suggesting that YMT inhibits osteoclast differentiation by suppressing RANKL-induced signaling molecules and transcription factors that affect the regulation of genes for osteoclast differentiation. As such, YMT may have therapeutic potential in bone diseases.

Samul-tang (SMT) is a traditional herbal medicine used for the treatment of ischemic brain and heart diseases. In this study, we investigated the molecular mechanism underlying the inhibitory effects of SMT on osteoclast differentiation induced by the receptor activator for nuclear factor κB ligand (RANKL) in RAW264.7 cells. SMT dose-dependently and significantly inhibited the RANKL-induced tartrate-resistant acid phosphatase (TRAP) activity and the formation of multi-nucleated osteoclasts. In addition, SMT significantly reduced the RANKL-induced mRNA expression of osteoclast differentiation-related genes that encode TRAP, c-src, matrix metalloproteinase (MMP)-9, cathepsin K, and the d2 isoform of vacuolar ATPase V(0) domain (ATPv0d2). SMT also inhibited the RANKL-induced activation of MAP kinases and NF-κB p65, and mRNA expression of transcription factors (Fra-2 and NFATc1). In conclusion, these results suggested that SMT suppresses the RANKL-induced activation of signaling molecules and transcription factors, which consequently inhibits osteoclast differentiation.

This study investigated quality characteristics of 4 year-old ginseng by enzymatic hydrolysis conditions to increase utilization. Ginseng was ground after steaming and was each treated with hydrolase A, B, C and D. When quality characteristics by the enzymes were examined, no significant difference was observed with pH of 5.5~5.6 and the sugar content of 4.0~4.33. The crude saponin content was the highest in ginseng treated with D, followed by B, C and A. The crude saponin, the reducing sugar and the total sugar contents increased until 0.3% (w/w) concentration in enzyme D with no significant difference by its concentration. Although active ingredients increased with time passage of hydrolysis, no significant change was found after three hours and the crude saponin content was the highest when ginseng was treated at . From these results, optimum conditions for 4 year-old ginseng were for 3 hours with 0.3% (w/w) enzyme D, and under these conditions the reducing sugar, the total sugar and the crude saponin contents recorded 18.11, 36.21 and 4.23 mg/g, respectively. Therefore, enzymatic hydrolysis was found to be effective in increasing active ingredients of 4 year-old ginseng with various usages expected.

Galgeun-tang (GGT) is a traditional Korean medication known to have a diaphoretic effect and to improve cerebral circulation. The aim of this study was to investigate the effects of GGT on osteoclast differentiation and bone loss in ovariectomized (OVX) rats compared to Galgeun-tang fermented by Bifidobacterium breve (designated as GFT). GGT significantly and dose-dependently inhibited the receptor activator for nuclear factor κB ligand (RANKL)-induced tartrate-resistant acid phosphatase (TRAP) activity and the formation of multinucleated osteoclasts in RAW264.7 cells. In addition, GGT significantly reduced RANKL-induced mRNA expression levels of TRAP, c-Src, and Cathepsin K. To examine the effect of GGT or GFT in OVX rats, we administered GGT or GFT (15 mL/kg/day) orally to OVX rats for 12 weeks. GGT administration significantly reduced body weight gain in OVX rats. GGT administration also significantly reduced total serum cholesterol, high-density lipoprotein (HDL)-cholesterol, low-density lipoprotein (LDL)-cholesterol, and triglyceride levels in OVX rats. GFT administration significantly reduced serum LDL-cholesterol and triglyceride levels in OVX rats. Moreover, administration of GGT, but not GFT, significantly increased bone mineral density of the femur, which is normally decreased in OVX rats. In conclusion, these results suggest that GGT could have the potential to decrease ovariectomy-induced increases in body weight and lipid content and could prevent bone loss through its inhibitory effect on osteoclast differentiation.

Ssanghwa-tang (SHT) is a traditional Korean herbal medicine widely prescribed to decrease fatigue following an illness. The purpose of this study was to investigate the effects of SHT on osteoclast differentiation in vitro, and on bone loss in ovariectomized (OVX) rats in vivo. SHT significantly reduced the receptor activator for the nuclear factor κB (NF-κB) ligand (RANKL)-induced tartrate-resistant acid phosphatase (TRAP) activity, and multinucleated osteoclast formation in RAW264.7 cells without affecting cell viability. In addition, SHT significantly attenuated RANKL-induced mRNA expression levels of c-Src and cathepsin K. To examine the in vivo effect of SHT on OVX-induced bone loss in OVX rats, we administered SHT (0.6 g/kg BID) orally to OVX rats for 12 weeks. SHT administration significantly blocked OVX-induced decrease of femoral bone mineral density (BMD) and femoral trabeculae in OVX rats. In conclusion, these results suggest that SHT treatment effectively prevents OVX-induced bone los...

Sinomenine is an alkaloid compound and a prominent anti-allergic agent found in the root of the climbing plant Sinomenium acutum. However, its effects on the bone marrow-derived mast cell (BMMC) mediated allergy and inflammation mechanism remain unknown. In this study, the biological effects of sinomenine were evaluated while focusing on its effects on the allergic mediator in PMA plus A23187-stimulated BMMCs. An investigation was also conducted to determine its effects on the production of several allergic mediators including interleukin-6 (IL-6), prostaglandin D(2) (PGD(2)), leukotriene C(4) (LTC(4)), β-Hexosaminidase (β-Hex), and cyclooxygenase-2 (COX-2) protein. The results revealed that sinomenine inhibited the PMA plus A23187-induced production of IL-6, PGD(2), LTC(4), β-Hex, and COX-2 protein. Taken together, these findings indicate that sinomenine has the potential for use in the treatment of allergy.

Angelica tenuissima Nakai is a widely used commodity in traditional medicine. Nevertheless, no study has been conducted on the antiviral and immune-modulatory properties of an aqueous extract of Angelica tenuissima Nakai. In the present study, we evaluated the antiviral activities and the mechanism of action of an aqueous extract of Angelica tenuissima Nakai both in vitro and in vivo. In vitro, an effective dose of Angelica tenuissima Nakai markedly inhibited the replication of Influenza A virus (PR8), Vesicular stomatitis virus (VSV), Herpes simplex virus (HSV), Coxsackie virus, and Enterovirus (EV-71) on epithelial (HEK293T/HeLa) and immune (RAW264.7) cells. Such inhibition can be described by the induction of the antiviral state in cells by antiviral, IFNrelated gene induction and secretion of IFNs and pro-inflammatory cytokines. In vivo, Angelica tenuissima Nakai treated BALB/c mice displayed higher survivability and lower lung viral titers when challenged with lethal doses of highly pathogenic influenza A subtypes (H1N1, H5N2, H7N3, and H9N2). We also found that Angelica tenuissima Nakai can induce the secretion of IL-6, IFN-λ, and local IgA in bronchoalveolar lavage fluid (BALF) of Angelica tenuissima Nakai treated mice, which correlating with the observed prophylactic effects. In HPLC analysis, we found the presence of several compounds in the aqueous fraction and among them; we evaluated antiviral properties of ferulic acid. Therefore, an extract of Angelica tenuissima Nakai and its components, including ferulic acid, play roles as immunomodulators and may be potential candidates for novel anti-viral/anti-influenza agents.