Combination therapy could be of use for the treatment of fungal infections, especially those caused by drug-resistant fungi. However, the methods and approaches used for data generation and result interpretation need further optimizing. The fractional inhibitory concentration index (FICI) is the most commonly used method, but it has several drawbacks in characterizing antifungal drug interaction. Alternatively, some new methods can be used such as the ⌬E model (difference between the predicted and measured fungal growth percentages) and the response surface approach, which uses the concentration-effect relationship over the whole concentration range instead of just the MIC. In the present study, in vitro interactions between tacrolimus (FK506) and three azoles-fluconazole (FLC), itraconazole (ITR), and voriconazole (VRC)-against Candida albicans were evaluated by the checkerboard microdilution method and time-killing test. The intensity of the interactions was determined by visual reading and the spectrophotometric method in a checkerboard assay, and the nature of the interactions was assessed by nonparametric models of FICI and ⌬E. Colony counting and colorimetric viable detection methods (2,3-bis {2-methoxy-4-nitro-5-[(sulfenylamino) carbonyl]-2H-tetrazolium hydroxide} [XTT] reduction test)were used for evaluating the combination antifungal effects over time. Synergistic and indifferent effects were found for the combination of FK506 and azoles against azole-sensitive strains, while strong synergy was found against azole-resistant strains analyzed by FICI. The ⌬E model gave more consistent results with FICI. The positive interactions were also confirmed by the time-killing test. Our findings suggest a potential role for combination therapy with calcineurin pathway inhibitors and azoles to augment activity against resistant C. albicans.

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Cyclosporin A showed potent synergism when combined with the three azoles, especially against azole-R C. albicans strains, and there was good agreement between various methods used in this study.

Recent studies have demonstrated that the histone deacetylation level was closely related to the genesis and development of tumors. Thus, activating histone acetyltransferases and/or suppressing histone deacetylases (HDACs) can become an approach for tumor chemotherapy. The histone acetylation regulation often results in the inhibition of cell proliferation, induction of cell apoptosis or differentiation, and cell cycle arrest in G1 phase. It has been demonstrated recently that the traditional anticonvulsant valproic acid was an efficient class I HDAC inhibitor (HDACI); however, its antitumor effect and mechanisms on gastric cancers so far has not been elucidated clearly. In the present study, gastric carcinoma cell lines BGC-823, HGC-27, and SGC-7901 were cultured with valproic acid (VPA) in vitro. The cell morphology was observed by invert microscope, the proliferation was detected by MTT assay, the apoptosis and cell cycle were analyzed by flow cytometry assay with Annexin V/PI and PI, the activities and protein expressions of Caspase 3, Caspase 8, Caspase 9 of BGC-823 cells were detected by spectrophotometry and indirect immunofluorescence technique, respectively. The protein expressions of Cyclin A, Cyclin D1, Cyclin E, P21(Waf/cip1) of BGC-823 cells were analyzed by indirect immunofluorescence assay, and messenger ribonucleic acid (mRNA) expressions were detected by RT-PCR assay. The results showed that the proliferation of three kinds of gastric carcinoma cells could be inhibited obviously by VPA, which was related to the apoptosis induction and cell cycle arrest in G1 phase. The intrinsic pathway (cytochrome C pathway) was chiefly involved in the mechanism of apoptosis, which was indicated by activation of Caspase 9 and Caspase 3. The extrinsic pathway was partially involved, with slight activation of Caspase 8. The mechanism underlying its effect on cell cycle arrest in G1 phase induction was due to the upregulation of P21(Waf/cip1), Mad1 expression and downregulation of Cyclin A, c-Myc expression.