Preventing Candida albicans from subverting host plasminogen for invasive infection treatment.

Candida albicans is a common fungal pathogen in humans that colonizes the skin and mucosal surfaces of the majority healthy individuals. How C. albicans disseminates into the bloodstream and causes life-threatening systemic infections in immunocompromised patients remains unclear. Plasminogen system activation can degrade a variety of structural proteins in vivo and is involved in several homeostatic processes. Here, for the first time, we characterized that C. albicans could capture and "subvert" host plasminogen to invade host epithelial cell surface barriers through cell-wall localized Eno1 protein. We found that the "subverted" plasminogen system plays an important role in development of invasive infection caused by C. albicans in mice. Base on this finding, we discovered a mouse monoclonal antibody (mAb) 12D9 targeting C. albicans Eno1, with high affinity to the 254FYKDGKYDL262 motif in α-helices 6, ß-sheet 6 (H6S6) loop and direct blocking activity for C. albicans capture host plasminogen. mAb 12D9 could prevent C. albicans from invading human epithelial and endothelial cells, and displayed antifungal activity and synergistic effect with anidulafungin or fluconazole in proof-of-concept in vivo studies, suggesting that blocking the function of cell surface Eno1 was effective for controlling invasive infection caused by Candida spp. In summary, our study provides the evidence of C. albicans invading host by "subverting" plasminogen system, suggesting a potential novel treatment strategy for invasive fungal infections.

Simultaneous determination of 5 major bioactive components in hugu capsule by HPLC-DAD.

OBJECTIVE: To develop a qualitative and quantitative determination of multiple components for quality control of Hugu Capsule, a composite prescription in TCM. METHODS: HPLC analysis was performed on a Kromasil C18 column (5.0 microm, 250 mm x 4.6 mm) by gradient elution with methanol-acetonitrile-1% glacial acetic acid as the mobile phase at a flow-rate of 0.8 mL/min,the detection wavelengths were set at 270, 283, 320 and 325 nm at the temperature of 30 degrees C. RESULTS: Five components: chlorogenic acid, tetrahydroxy stilbene glucoside, ferulaic acid, naringin and icariin were simultaneously analyzed in this study. The calibration curves were exhibited linear regressions of at least r > 0.9992. The injection precision,the intra-day precisions and the analysis repeatability were evaluated with the RSD values were all less than 5%. The mean recoveries of the five components were ranged from 97.4% to 99.4%, and RSD values all were less than 1.72%. CONCLUSION: This method is found to be convenient, fast, accurate, and is applicable to analyze the multi-constituents in Hugu Capsule.

Protective effect of total alkaloids of Sophora alopecuroides on dextran sulfate sodium-induced chronic colitis.

OBJECTIVE: To investigate the effect of total alkaloids of Sophora alopecuroides (TASA) on dextran sulfate sodium (DSS)-induced colitis in mice. METHODS: Chronic experimental colitis was induced by administration of 4 cycles of 4% DSS. Fifty mice were randomly distributed into 4 groups (normal, DSS, DSS/high-dose TASA, and DSS/low-dose TASA groups) by a random number table with body weight stratification. Mice in the normal group (n=11) and DSS-induced colitis control group (n=15) received control treatment of 20 mL/kg distilled water; DSS plus TASA high- and low-dose groups (n=12 each) were treated with TASA solution (20 mL/kg) at the doses of 60 mg/kg and 30 mg/kg, respectively. The severity of colitis was assessed on the basis of clinical signs, colon length, and histology scores. Moreover, secretory immunoglobulin A (sIgA) and haptoglobin (HP) were analyzed by enzyme linked immunosorbent assay; intercellular adhesion molecule 1 (ICAM-1) and macrophage-migration inhibitory factor (MIF) gene expressions were analyzed by quantitative reverse transcriptase realtime polymerase chain reaction (qRT-PCR) using SYBA green I; and nuclear factor κ B (NF-κ B) expression and activation and p65 interaction with the promoter of ICAM-1 gene were assessed by Western blotting and chromatin immunoprecipitation assay. RESULTS: TASA administration significantly attenuated the damage and substantially reduced HP elevation and maintained the level of cecum sIgA. TASA inhibited the ICAM-1 gene expression and had no effect on MIF gene expression. Also, TASA was able to reduce phospho-I κ B α (p-I κ B α) protein expression; however, it had no effect on the activation of I κ B kinase α (IKK α) and inhibitor of NF-κ B α (I κ B α). Moreover, TASA inhibited the p65 recruitment to the ICAM-1 gene promoter. CONCLUSIONS: TASA had a protective effect on DSS-induced colitis. Such effect may be associated with its inhibition of NF-κ B activation and blockade of NF-κ B-regulated transcription activation of proinflammatory mediator gene.

[Analysis on the HPLC chromatographic fingerprint of gegen qinlian decoction].

OBJECTIVE: To investigate the multiple bioactive constituents in Gegen Qinlian decoction and evaluate its bioactivity and quality control. METHODS: A high performance liquid chromatography with diode array detection (HPLC-DAD) method for the simultaneous qualitative determination of multiple components was developed. The separation was performed on a Kromasil C18 column by gradient elution with acetonitrile and ammonium acetate (containing 0.3% triethylamine, and adjusted pH 4. 3 using 1% glacial acetic acid) as the mobile phase at a flow-rate of 0.7 mL/min, the column temperature was maintained at 30 degrees C, HPLC chromatographic comparison between Gegen Qinlian decoction, four medicinal material and 14 components were carried out to investigate components in the composite formula. RESULTS: 23 characteristic peaks were identified and could be dated back to four medicinal materials from Gegen Qinlian decoction, in which 12 peaks were identifed by standards. CONCLUSION: This readily available, low-cost and reliable HPLC-DAD method can be used for simultaneous analysis of multiple bioactive constituents of Gegen Qinlian decoction and will improve the quality control of Gegen Qinlian decoction.

[Effect of pectins of different degree of esterification on in-vitro sophoridine release of hydrophilic matrix tablets containing total alkaloids of Sophora alopecuroides].

OBJECTIVE: To prepare colon-targetting tablets of total alkaloids of Sophora alopecuroides and evaluate the effect of pectins of different degree of esterification (DE) on sophoridine release profiles in-vitro. METHOD: Wet granulation technique was employed to prepare petin-based matrix tablets, then tablets were coated the optimal formulation with Kollicoat MAE 30 DP based on the optimal formulation and analysed their release. RESULT: Coated formulation E could target total alkaloids of S. alopecuroides to colon and various DE of pectin exerted different effects on sophoridine release. The release of low DE pectin-based matrix tablets coating with Kollicoat MAE 30 DP approximatedly fitted zere-order eqution, which was erosion depended. CONCLUSION: Low DE pectin-based matrix tablet coating with Kollicoat MAE 30 DP can deliver sophoridine to colon, hence improve the effectiveness of sophoridine.