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2296 J. Med. Plants Res. Table 1. Antifungal activity of extract of Coptidis Rhizoma against Candida species. Species Clear zone (mm) contents of extract (μg/ml) 10 50 100 150 200 C. albicans - - - - 3 C. tropicalis - 2 3 6 9 C. glabrata - 3 4 4 5 C. parapsilensis - - - - - C. utils - - - - - -: Not detected. DMSO was added to dissolve the formazan crystals. The absorbance was measured at 560 nm with a spectrophotometer. RESULTS AND DISCUSSION Anticandidial activity of Coptidis Rhizoma extract was determined against five different Candida sp. by agar diffusion assay. The extract of Coptidis Rhizoma showed antifungal activity against Candida albicans at 200 μg/ml, C. tropicalis and C. glabrata at 50 μg/ml. The extract showed best antifungal activity against C. glabrata at 50 μg/ml (Table 1). A compound 1 was separated by TLC analysis from Coptidis Rhizoma extract, and the above compound 1 was confirmed as berberine by HPLC analysis (Figure 1). The MIC (minimum inhibition concentration) of the compound 1 and berberine were Figure 1. HPLC analysis of compound 1 isolated from coptidis rhizoma. determined and compared. The MIC values of the compound 1 and berberine were 24 and 48 μg/ml against C. tropicalis and C. glabrata, and 30 and 50 μg/ml against C. tropicalis and C. glabrata (Table 2). The difference of MIC values of both compounds was supposed an error of weighting materials. The IC50 values of the compound 1 were 58 μM at 24 h and 40 μM at 48 h by MTT assay of cell viability using HaCaT cell line (Figure 2). The above results indicated that the compound 1, berberine, was a good candidate to inhibit the propagation of C. tropicalis and C. glabrata, even in human skin. Candidiasis has been the most common fungal infection in HIV infected individuals and patients in hospital. Although Candida spp. resistant to antifungal agents have been a rarity until three decades ago, resistance of Candida spp. was spread in a serious infection disease of the hospital environment. Although
Table 2. MIC of a compound 1 from coptidis rhizoma. Species Compound 1 (μg/ml) Berberine (μg/ml) Amphotericin B (μg/ml) C. albicans - - 0.2 C. tropicalis 24 30 0.4 C. glabrata 48 50 0.4 C. parapsilensis - - 0.4 C. utils - - 0.4 - : Not detected. Cell viability ( IC 50 value(M) ) 120 100 80 60 40 20 0 24 h 48 h Amphotericin B 24hr 48hr Berberin Figure 2. IC50 values for viability on HaCaT cells. Growth inhibition of HaCaT cell after treatment with berberine and Amphotericin B. Cells were plated at 1x10 4 cell/well per 96 well plate, and incubated for 24 h. The cells treated with variable concentrations of berberine for 24 and 48 h, and amphotericin B for 24 h and the viability were measured by MTT assay. The data represent means ±SD of five independent experiments. uncommon, this outbreak should serve as a warning that multi-resistance of Candida strains may develop and spread within the hospital environment (Bodey, 1988; Schaberg et al., 1991; Masia and Gutierrez, 2002). In this work, we did seek what a kind of the extract from Coptidis Rhizoma was responsible for the antifungal effect. It is suggested that the compound 1, berberine, can be applied as a phytomedicine for treatment of the Candidal infection (Williamson, 2001; Han and Lee, 2005). ACKNOWLEDGEMENT This research was supported by the <strong>Academic</strong> Research Fund of Hoseo University in 2010 (2010-0131). REFERENCES Kim et al. 2297 Abi-Said D, Anaissie E, Uzun O, Raad I, Pinzcowski H, Vartivarian S (1997). The epidemiology of hematogenous candidiasis caused by different Candida species. Clin. Infect. Dis., 24: 1122-1128. Blumberg EA, Reboli AC (1996). Failure of systemic empirical treatment with amphotericin B to prevent candidemia in neutropenic patients with cancer. Clin. Infect. Dis., 22: 462-466. Bodey GP (1988). The emergence of fungi as major hospital pathogens. J. Hosp. Infect., 11: 411-426. Fonos V, Cataldi L (2000). Amphotericin B-induced nephrotoxicity. J. Chemother., 12: 463-470. Gallis HA, Drew RH, Pickard WW (2000). Amphotericin B: 30 years of clinical experience. Rev. Infect. Dis., 12: 308-329. Girmenia C, Martino P (1998). Fluconazole and the changing epidemiology of candidemia. Clin. Infect Dis., 27: 234. Han Y, Lee JH (2005). Berberine synergy with amphotericin B against disseminated candidiasis in mice. Biol. Pharm. Bull., 28: 541-544. Masia CM, Gutierrez RF (2002). Antifungal drug resistance to azoles
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Journal of Medicinal Plants Researc
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Editors Prof. Akah Peter Achunike E
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Electronic submission of manuscript
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Fees and Charges: Authors are requi
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Table of Contents: Volume 6 Number
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Figure 1. Chuanxiong Rhizoma (http:
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Table 1. Identification of main pea
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information on plants used for the
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Table 1. Levels of independent vari
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Arbutin (mg/g) Co-solvent Figure 1.
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Arbutin (mg/g) Extraction pressure
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Table 3. Arbuitn content of Asian p
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Arbutin (mg/g) Extraction pressure
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Arbutin (mg/g) Extraction temperatu
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Table 1. Chemical composition of es
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Darjazi 2369 Table 2. Statistical a
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Table 3. Correlation matrix (number
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Table 1. Precision test. extraction
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Figure 2. Effect of different alcoh
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Table 6. Results of ANOVA. Nie et a
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Table 2. Plant data and MIC values
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exhibiting the polar extracts of P.
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Table 3. Contd. AcOEt ++ ++ ++ - Me
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Figure 1. Map of Ladakh region of I
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Warghat et al. 2391 Table 3. Summar
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Table 5. Inter-Population genetic i
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Excoffier L, Smouse PE, Quattro JM
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ice residue is an ideal raw materia
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DPPH• scavenging ratio(%) OD valu
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scavenging effect on free radical a
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our new CL system. Fenton reaction
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1 2 Time (s) Figure 2. Kinetic curv
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Wong et al., 2006). It can be seen
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Pan YM, Liang Y, Wang HS, Liang M (
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2422 J. Med. Plants Res. close cano
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2442 J. Med. Plants Res. Stability
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UPCOMING CONFERENCES 15th European
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