目的 研究汉防己甲素(tetrandrine，TET)对联苯苄唑(bifonazole，BFZ)治疗须癣毛癣菌所致豚鼠皮肤癣菌病是否有增效作用.方法 以须癣毛癣菌构建豚鼠皮肤癣菌病模型，将造模成功的20只豚鼠按随机数字表法等分为4组，进行双盲实验，分别涂用复方BFZ(含1%BFZ和1% TET)、1%BFZ、1%TET和基质，1次/d，共14天，进行皮损评分、病原学和病理学检查，分析4组药物间疗效差异.结果 用药期间，4组皮损评分差异有统计学意义(F=93.49，P=0.000);从用药第3天起，复方BFZ组和1%BFZ组皮损评分逐日下降，前者皮损评 分持续低于后者，差异有统计学意义(P=0.000);1%TET组和基质组皮损评分差异无统计学意义(P=1.000).复方BFZ组皮损真菌学检查比 1%BFZ组转阴早2天，炎症反应轻于其他3组.结论 TET对BFZ治疗须癣毛癣菌所致豚鼠皮肤癣菌病有增效作用.
ABSTRACT The aim of present work was to develop and evaluate solid lipid nanoparticles (SLNs) based gel for topical delivery of antifungal agent. Bifonazole (BFZ) is an imidazole antifungal agent having broad spectrum activity against dermatophytes， moulds， yeasts， fungi and some gram positive bacteria. BFZ SLNs systems were developed by melt emulsification followed by ultrasonication technique using Precirol ATO5 as a solid lipid and Tween 80 as a surfactant. Developed SLNs were evaluated for particle size， polydispersity index (PI)， entrapment efficiency (EE) and drug release profiles. Process and formulation parameters were optimized. Differential scanning calorimetry (DSC) and X-ray diffraction (XRD) studies were carried out on SLNs to mark the changes in the drug and lipid modifications. The BFZ SLNs based gels were prepared using Carbopol 940 as a gelling agent. The SLNs based gels were evaluated for rheological parameters， in vitro drug release and permeation studies. In vitro antifungal study suggested that the SLNs based gel was more effective in inhibiting growth of Candida albicans. Thus the study concludes that SLNs based gel of BFZ gives a sustained release profile of BFZ and has the potential for treatment of topical fungal infections.
[In vitro antifungal susceptibility profile of Scopulariopsis brevicaulis isolated from onychomycosis].
We studied the in vitro antifungal activity profile of amorolfine (AMR)， bifonazole (BFZ)， clotrimazole (CLZ)， econazole (ECZ)， fluconazole (FNZ)， itraconazole (ITZ)， ketoconazole (KTZ)， miconazole (MNZ)， oxiconazole (OXZ)， tioconazole (TCZ) and terbinafine (TRB) against 26 clinical isolates of Scopulariopsis brevicaulis from patients with onychomycosis by means of an standardized microdilution method. Although this opportunistic filamentous fungi was reported as resistant to several broad-spectrum antifungals agents， obtained data shows a better fungistatic in vitro activity of AMR， OXZ and TRB (0.08， 0.3， and 0.35 mg/L， respectively) in comparison to that of CLZ (0.47 mg/L)， ECZ (1.48 mg/L)， MNZ (1.56 mg/L， BFZ (2.8 mg/L)， TCZ (3.33 mg/L)， KTZ (3.73 mg/L). FNZ (178.47 mg/L) and ITZ (4.7 mg/L) showed a reduced in vitro antifungal activity against S. brevicaulis. Obtained MICs show the low in vitro antifungal susceptibility of S. brevicaulis to topical drugs for onychomycosis management， with exceptions (AMR， OZX and TRB).
There have been the current Japanese data on susceptibility testing forCandidaisolates from vaginal candidiasis. Thein vitroactivities of therapeutic antifungal drugs for vulvovaginal candidiasis (VVC); miconazole (MCZ)， itraconazole (ITCZ)， fluconazole (FLCZ)， clotrimazole (CTZ)， oxiconazole (OCZ)， isoconazole (ICZ) and bifonazole (BFZ) against vaginal isolates. Fifty-four strainsCandida albicansand 19 strains ofCandida glabratawere evaluated using a broth microdilution method specified by Clinical Laboratories Standard Institute (CLSI) document M27-A3. The MIC90of each drug， MCZ， ITCZ， FLCZ， CTZ， OCZ， ICZ and BFZ， againstC. albicansandC. glabrataisolates were 0.25， 0.12， 1， 0.06， 0.12， 0.12 and 1 μg/ml and 1， 1， 8， 0.5， 0.25， 0.5 and 1 μg/ml respectively. The activities of these drugs， except for BFZ， againstC. glabratawere lower than that of C. albicans. There was one azole-resistant isolate inC. glabrataof which MIC of FLCZ is > 64 μg/ml and this isolate had cross resistance to other antifungal drugs tested. These results suggest that antifungal drugs for treatment of VVC continues to have potent antifungal activities againstC. albicansandC. glabrataisolates from vaginitis. CTZ， OCZ and ICZ susceptibility of FLCZ low susceptibilityC. glabrataare relatively higher than MCZ， ITCZ and FLCZ.
Comparison of Superficial Mycosis Treatment using Butenafine and Bifonazole nitrate Clinical Efficacy
Abstract Superficial fungal infections are commonly encountered by the physician. And the continuously changing epidemiology of invasive fungal infections results in the need for an expanded armamentarium of antifungal therapies. This study was designed to evaluate the safety and efficacy of Butenafine (BTF) versus Bifonazole (BFZ) in the treatment of superficial mycosis in a randomized， double-blind， parallel-group trial. Of 96 patients， 48 applied (BTF) cream and 48 applied (BFZ) cream for 2 weeks to tinea versicolor， corporis and cruris treat， while tinea of feet & hands was treated for 4 weeks duration. Efficacy was assessed after the end of treatment and 2 weeks later. At the end of therapy， we find somewhat more patients using (BTF) than using (BFZ) had a mycologic cure ((BTF)， 87.5%; (BFZ) 83.3%) and effective clinical response ((BTF)， 91.7%; (BFZ)， 83.3%). (BTF) provides rapid and persistent antifungal activity and symptom relief in patients with superficial mycosis during treatment. And patients continued to improve for at least 2 weeks after treatment. The Rates of mycologic cure and effective treatment with (BTF) were higher than with (BFZ) at cessation of treatment and 2 weeks later. However， no significant difference found between the two drugs (p> 0.05).
Binding of diuretic antihypertensive bendroflumethiazide to human serum albumin studied by 19 F nuclear magnetic resonance method
Simultaneous specific and nonspecific binding of bendroflumethiazide (BFZ) to human serum albumin (HSA) and concentration profile of BFZ in HSA buffer (pH 7.40) solution were investigated by 19 F nuclear magnetic resonance (NMR) method. The 19 F NMR spectrum of BFZ (200μM) in a buffer solution showed a sharp signal of its CF 3 group at 17.8ppm from the reference trifluoroethanol. Addition of 0.60mM HSA to the sample solution caused the CF 3 signal splitting into three broadened peaks at 18.4 (A)， 17.9 (B) and 17.4ppm (C). By its chemical shift and spectral behavior， B was assigned to unbound BFZ. Competition experiments with Site I and II ligands lead to C being assigned to Site II bound BFZ. However， the peak intensity (areas) of A was not reduced by these ligands， suggesting that A arises from nonspecific binding. Using the peak intensities at several total concentrations of BFZ， Scatchard plot was performed. The plot for A provided a straight line parallel to the x -axis confirming nonspecific binding and that for C was consistent with specific binding. The binding constants for nonspecific and specific Site II binding were 1.02 and 1.00×10 4 (M −1 ) ( n =1.1)， respectively. The presence of 0.10M Cl − in the sample solution affected the binding constant of Site II binding， but not that of nonspecific binding. The concentration profile of BFZ calculated using the binding constants revealed that nonspecific binding is more effective than Site II binding for the binding of BFZ to HSA. It was also confirmed that considerable amounts of BFZ liberated from Site II by the Site II ligands or Cl − ions bind again nonspecifically.
[Influence of the ecological group on the in vitro antifungal susceptibility of dermatophytic fungi].
BACKGROUND： Dermatophytes can be divided into geophilic (soil)， zoophilic (animals) and anthropophilic (humans) strains， depending on the source of the keratin that they use for nutritional purposes. AIMS： The in vitro susceptibility of clinical isolates of dermatophyte fungi has been studied in the 3 ecological groups with several antifungal agents for the topical management of dermatophytoses in order to determine their relationship with the ecological group. METHODS： A standardised dilution micromethod in a liquid medium was used for the determination of the in vitro antifungal activity of 9 topical antifungal drugs： amorolfine (AMR)， bifonazole (BFZ)， clotrimazole， econazole， ketoconazole， miconazole， oxiconazole， terbinafine (TRB) and tioconazole. The in vitro activity was obtained against 124 clinical isolates of dermatophyte moulds from the anthropophilic， zoophilic and geophilic ecological groups. RESULTS： The in vitro antifungal activity was different depending on the ecological group， although a species-dependent profile was also observed. CONCLUSIONS： Azole derivatives showed a similar antifungal profile， being more active against anthropophilic dermatophytes > zoophilic > geophilic. Activity of TRB and AMR was different from that of azole derivatives (zoophilic > anthropophilic > geophilic). A higher in vitro antifungal activity against the 3 ecological groups was observed with TRB and AMR， whilst BFZ was the less active drug. Copyright © 2012 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.
Objective To investigate the synergistic effect of tetrandrine (TET) to bifonazole(BFZ) treatment of dermatophytosis in guinea pigs infected with Trichophyton mentagrophytes. Methods Establishing the dermatophytosis model in guinea pigs with Trichophyton mentagrophytes infection. The 20 infected animals were randomly and equally divided into 4 groups to conduct double-blind experiment and were applied with BFZ compound cream(containing 1%BFZ and 1%TET)，1%BFZ cream， 1%TET cream and matrix cream once per day for successive 14 days respectively. The alterations in the skin lesion scores， mycological and pathology were observed to analyze the efficacy differences between four groups of drugs. Results During the treatment， there were obvious statistical difference (F=93.49，P=0.000)between the skin lesion scores in the four groups; the skin lesion scores of BFZ compound cream group and 1%BFZ cream group started to fall gradually from the 3th day， the former was significantly more effective than the latter， with obviously statistical difference (P=0.000); there was no statistical difference (P=1.000) between the skin lesion scores of 1%TET cream group and matrix cream group. In the BFZ compound cream group， the skin lesion scores became negative 2 days earlier than the 1%BFZ cream group， and inflammatory reaction was slighter than the other 3 groups. Conclusion There is synergistic effect of TET on BFZ treatment of dermatophytosis in guinea pigs infected with Trichophyton mentagrophytes.
A minority of patients on long term treatment with thiazides or beta blockers develop diabetes， and guidelines suggest the risk from such drugs is additive. Our aims were to estimate [i] whether anincrease in 2-h glucose in the oral glucose tolerance test (OGTT) is detectable after 4 weeks of high-dose treatment， and [ii] whether any change is amplified by combining the classes.42 patients with essential hypertension， aged 35-74 (median 59)， 30 male， 12 female， participated in a(4 vs 0 weeks)， 0.013 (BFZ vs placebo). There was no change in 30-min insulin.
In this study， molecular modelling analyses based on molecular mechanics， semi-empirical (PM3) and DFT (at B3LYP/6-31G* level) calculations have been carried out to obtain insight into their toxicity. The results of the analyses show that LLZ， LCZ and BFZ have LUMO-HOMO energy differences ranging from 4.5 to 4.9 eV， indicating that the compounds would be moderately inert with BFZ being the most inert one. The molecular surfaces of all the compounds are found to possess significant amounts of positively charged electron-deficient regions so that they may be subject to nucleophilic attacks by glutathione and nucleobases in DNA， thus causing cellular toxicity due to glutathione depletion and DNA damage due to oxidation of nucleobases. However， because of kinetic inertness of the molecules， the rates of such adverse reactions are expected to be low.