Antifungal Activities of cis-trans Citral Isomers against Trichophyton rubrum with ERG6 as a Potential Target.

Trichophyton rubrum causes ringworm worldwide. Citral (CIT), extracted from Pectis plants, is a monoterpene and naturally composed of geometric isomers neral (cis-citral) and geranial (trans-citral). CIT has promising antifungal activities and ergosterol biosynthesis inhibition effects against several pathogenic fungi. However, no study has focused on neral and geranial against T. rubrum, which hinders the clinical application of CIT. This study aimed to compare antifungal activities of neral and geranial and preliminarily elucidate their ergosterol biosynthesis inhibition mechanism against T. rubrum. Herein, the disc diffusion assays, cellular leakage measurement, flow cytometry, SEM/TEM observation, sterol quantification, and sterol pattern change analyses were employed. The results showed geranial exhibited larger inhibition zones (p < 0.01 or 0.05), higher cellular leakage rates (p < 0.01), increased conidia with damaged membranes (p < 0.01) within 24 h, more distinct shriveled mycelium in SEM, prominent cellular material leakage, membrane damage, and morphological changes in TEM. Furthermore, geranial possessed more promising ergosterol biosynthesis inhibition effects than neral, and both induced the synthesis of 7-Dehydrodesmosterol and Cholesta-5,7,22,24-tetraen-3ß-ol, which represented marker sterols when ERG6 was affected. These results suggest geranial is more potent than neral against T. rubrum, and both inhibit ergosterol biosynthesis by affecting ERG6.

[Preparation of O/W ginseng saponins-based nanoemulsion and its amplified immune response].

OBJECTIVE: To prepare an O/W ginseng saponins-based nanoemulsion and investigate its amplified immune response. METHOD: The formulation of ginseng saponins-based nanoemulsion was optimized via the range of nanoemulsion zone in phase diagrams and the solubility of ginseng saponins. Its physicochemical properties were investigated, including morphology, particle size distribution, pH, viscosity and stability. Ginseng saponins-based nanoemulsion as adjuvant was co-administrated with a model antigen ovalbumin (OVA) in mice. Two weeks after the boosting, the serum levels of OVA-specific antibody and its isotypes were determined. RESULT: The optimized ginseng saponins-based nanoemulsion formulation consisted of ginseng saponins, IPM, Cremophor RH 40, glycerol and water (with the weight ratio of 2 : 4 : 17.8 : 17.8 : 58.4), which was a light yellow fluid. The shape of droplets was spherical under transmission electron microscopy with an average diameter of 72.20 nm and a polydispersity index of 0.052. The viscosity and pH value of it were 4.20 s and 6.02, respectively. And it showed good stability. When co-administered with OVA, no obvious side effects were observed in the mice immunized with ginseng saponin-based nanoemulsion. The serum levels of IgG, IgG1 and IgG2a antibody in the group of ginseng saponin-based nanoemulsion immunized mice was significantly increased compared to the groups of OVA and the saline solution of ginseng saponin. Compared with the adjuvant aluminium hydroxide, the serum levels of IgG and IgG1 antibodys in the groups of ginseng saponins-based nanoemulsion had no significant difference, but the level of IgG2a was obviously higher. CONCLUSION: ginseng saponin-based nanoemulsion could amplify the Th1 and Th2 immune responses, and can be used as the vaccine adjuvant.

[Effects of vinblastine nanoparticles on growth and apoptosis of glioma cell line C6].

OBJECTIVE: To compare effects of vinblastine (VLB) nanoparticles (NPS) and VLB physiologic saline solution on inhibiting glioma cell lines C6 growth and inducting its apoptosis. METHOD: Glioma cell lines C6 were respectively treated with 500 micro x L(-1) VLB NPS and VLB physiologic saline solution for 7 days. Amount of cells were counted by blood cell counting chamber. Glioma C6 growth curve was draw according to cells amount. Clone formation rate of glioma C6 was detected after 500 microg x L(-1) VLB NPS and VLB physiologic saline solution incubation for 2 weeks. In addition, the whole morphology of glioma C6 were observed by inverted microscope and inverted fluorescence microscope after 500 microg x L(-1) VLB NPS and VLB physiologic saline solution incubation for 48 h. RESULT: Entrapment of VLB in NPS may significantly inhibit glioma cells C6 growth from 2 to 7 days compared with VLB physiologic saline solution in the same dose (P < 0.05). Clone formation rate of glioma C6 in VLB physiologic saline solution group is 1. 3 times better than VLB NPS. The difference between VLB NPS and VLB physiologic saline solution is significant (P < 0.05). Results of morphology change indicated glioma cells C6 with the VLB NPS treatment were intermediate or end stage, missed structure integrality. Amount of cells was distinctly decreased, and apoptosis cells number was apparently increased compared with VLB physiologic saline solution group. CONCLUSION: VLB NPS have stronger cytotoxicity to glioma cells line C6 compared with VLB physiologic saline solution in the same dose. NPS may be effective as promising carrier for the transport of VLB into the glioma cells.

[Effect of vinblastine nanoparticles on antiproliferation in human glioma cell lines BT325].

OBJECTIVE: To compare antiproliferation effects of vinblastine nanopraticles and vinblastine water solution in human glioma cell lines BT325. METHOD: Vinblastine nanoparticles were prepared by emulsion polymerization process and using dextran as a stabilizing agent. It was characterized by means of morphology, size, drug entrapment efficiency and loading efficiency. Human glioma cell lines BT325 were treated with different concentrations of vinblastine nanoparticles and vinblastine water solution for 48 h, Antiproliferation effect was measured by MTT method. Morphological changes were observed by inverted microscope, transmission electron microscope and scanning electron microscope. RESULT: Mean diameter of VLB-PBCA-NP was about 74.4 nm, and drug entrapment efficiency and loading efficiency was 78.47% and 39.24%, respectively. Cell growth inhibition rate of vinblastine nanoparticles group and vinblastine water solution group in a concentration range (5-5 000 g x L(-1)) for 48 h was 41%, 49%, 73%, 83% and 28%, 33%, 54%, 60% respectively. Entrapment of VLB in NPS may distinctly degrade absorbency as compared to free drugs. Glioma cell BT325 which treated with VLB water solution were initial stage of apoptosis, and apoptosis body were forming. But VLB NPS-treated BT325 cells were intermediate or end stage, and missed structure integrality. CONCLUSION: VLB-PBCA-NP and VLB water solution could inhibit the growth of human glioma cell lines BT325, and VLB nanoparticles have stronger inhibition effect compared with VLB water solution in the same dose. PBCA may be effective as promising carrier for the transport of vinblastine into the glioma cells.