阿托伐他汀的肝毒性机制研究

目的 研究阿托伐他汀肝毒性损伤作用及机制。方法 将24只Wistar han雄鼠分为对照组和阿托伐他汀低（68.5mg/kg）、高剂量组（205.5 mg/kg）,按照10 mL/kg的药液体积给药,溶媒对照组ig等体积5% CMC-Na,连续ig 28 d。检测血清中天门冬氨酸氨基转移酶（AST）、丙氨酸氨基转移酶（ALT）、碱性磷酸酶（ALP）、尿素氮（BUN）和血肌酐（CRE）的含量,HE染色观察肝组织病理。在体外,HepG2细胞经传代培养后,给予阿托伐他汀干预24 h,检测细胞存活率,丙二醛（MDA）水平、Na⁺-K⁺-ATP酶和Ca²⁺-Mg²⁺-ATP酶活性及线粒体膜电位。结果 与对照组比较,阿托伐他汀高剂量组大鼠肝细胞弥漫性肿胀,核分裂多见,部分肝细胞极性消失,排列紊乱（P<0.05）。与对照组比较,阿托伐他汀高剂量组给药后血清中ALT和AST显著升高（P<0.05、0.01）。在体外,与对照组比较,阿托伐他汀125、250、500 μmol/L能明显抑制细胞存活率（P<0.05、0.001）。与对照组比较,阿托伐他汀500 μmol/L HepG2细胞MDA含量明显升高（P<0.01）。与对照组比较,阿托伐他汀125 μmol/L能使Na⁺-K⁺-ATP酶活性增强,500 μmol/L使Na⁺-K⁺-ATP酶活性降低（P<0.001）。与对照组比较,阿托伐他汀125、250、500 μmol/L均能使能使Ca²⁺-Mg²⁺-ATP酶活性降低（P<0.01,0.001）。与对照组比较,阿托伐他汀125、250、500 μmol/L均能降低线粒体膜电位（P<0.001）。结论 阿托伐他汀高剂量可导致肝组织损伤,其毒性作用通过破坏细胞的线粒体膜电位,抑制Na⁺-K⁺-ATP酶、Ca²⁺-Mg²⁺-ATP酶活性,细胞膜脂质过氧化,从而破坏细胞内微环境的平衡,导致细胞凋亡和坏死。     

半枝莲总黄酮调控MIIP对人胃腺癌细胞AGS增殖、凋亡和放疗敏感性的作用及机制研究

目的 研究半枝莲总黄酮（TF-SB）对人胃腺癌细胞AGS增殖、凋亡和放疗敏感性的影响和分子机制。方法 采用细胞计数试剂盒（CCK-8）法检测TF-SB（0、25、50、100、200 μg·mL⁻¹）作用24 h对AGS细胞存活率的影响。将AGS细胞分为对照组、溶剂对照（含相同浓度的甲醇）组、TF-SB（100 μg·mL⁻¹）组、质粒空载体（pcDNA,200 nmol·L⁻¹）组、迁移侵袭抑制蛋白（MIIP）过表达质粒（pcDNA-MIIP,200 nmol·L⁻¹）组、TF-SB（100 μg·mL⁻¹）＋小干扰RNA对照（si-con,200 nmol·L⁻¹）组、TF-SB（100 μg·mL⁻¹）＋MIIP小干扰RNA（si-MIIP,200 nmol·L⁻¹）组,转染质粒后TF-SB处理24 h,CCK-8法检测细胞存活率;流式细胞术检测细胞凋亡;Western blotting法检测B细胞淋巴瘤-xl（Bcl-xl）、裂解的半胱氨酸天冬氨酸蛋白酶（cleaved Caspase-3）、MIIP表达;不同照射量X射线（0、2、4、6、8 Gy）照射后,克隆形成实验检测细胞存活率,并绘制单击多靶模型拟合曲线,Western blotting法检测γ-H2AX蛋白表达。结果 与TF-SB 0 μg·mL⁻¹组比较,TFSB 25、50、100、200 μg·mL⁻¹组AGS细胞存活率显著降低（P＜0.05）;与对照组及溶剂对照组比较,TF-SB 100 μg·mL⁻¹组AGS细胞凋亡率显著升高（P＜0.05）,Bcl-xl蛋白表达显著降低（P＜ 0.05）,cleaved Caspase-3和MIIP蛋白表达显著升高（P＜0.05）;放疗后,TF-SB组细胞存活率显著降低（P＜0.05）,γ-H2AX蛋白表达显著升高（P＜0.05）,放疗敏感性增加。与pcDNA组比较,pcDNA-MIIP组AGS细胞凋亡率显著增加,细胞存活率显著降低,Bcl-xl蛋白表达显著降低,cleavedCaspase-3蛋白表达增加（P＜0.05）;放疗后,pcDNA-MIIP组细胞存活率显著降低（P＜0.05）,γ-H2AX蛋白表达显著升高（P＜0.05）,敏感性增加。与TF-SB＋si-con组比较,TF-SB＋si-MIIP组AGS细胞凋亡率显著降低,细胞存活率显著增加,Bcl-xl蛋白表达显著增加,cleaved Caspase-3蛋白表达显著降低（P＜0.05）;放疗后,TF-SB＋si-MIIP组细胞存活率显著升高（P＜0.05）,γ-H2AX蛋白表达显著降低（P＜0.05）,敏感性降低。结论 TF-SB通过上调MIIP可抑制AGS细胞增殖,诱导细胞凋亡,提高其放疗敏感性。     

新绿原酸抑制人宫颈癌HeLa细胞迁移的实验研究

目的 考察新绿原酸对宫颈癌HeLa细胞体外迁移模型和斑马鱼移植瘤模型的影响。方法 MTT法检测新绿原酸（20、40、80、160、320 μmol/L）对HeLa细胞增殖的作用;应用宫颈癌细胞HeLa划痕愈合模型和Transwell实验考察新绿原酸（28、56、113 μmol/L）对体外迁移作用的影响;同时利用构建的绿色荧光蛋白（GFP）的HeLa细胞系建立斑马鱼移植瘤模型,考察新绿原酸（56、113、226 μmol/L）对体内移植瘤模型细胞迁移的作用;Western blotting法检测新绿原酸对内皮生长因子（EGF）诱导的HeLa细胞中表皮生长因子受体（EGFR）、p-EGFR蛋白表达的影响。结果 MTT结果显示,新绿原酸在160 μmol/L及以下浓度对HeLa细胞增殖无明显影响;与对照组比较,56、113 μmol/L新绿原酸显著抑制划痕愈合,抑制率分别为49.3%（P<0.05）和64.1%（P<0.01）;Transwell实验结果提示,113 μmol/L新绿原酸具有显著抑制HeLa细胞向下层迁移的作用（P<0.05）;与模型组比较,113、226 μmol/L新绿原酸可显著减少斑马鱼移植瘤模型的HeLa细胞的迁移面积（P<0.05、0.01）;Western blotting结果显示,与EGF组比较,新绿原酸对EGFR表达和磷酸化有明显抑制作用,56、113 μmol/L浓度组差异显著（P<0.01）。结论 在未影响HeLa细胞增殖的浓度下,新绿原酸具有抑制宫颈癌HeLa细胞迁移的作用,其作用机制与抑制EGFR的表达有关。     

五味子乙素通过ROS介导内质网应激诱导人乳腺癌MDA-MB-231细胞凋亡的研究

目的 研究五味子乙素对人乳腺癌MDA-MB-231细胞凋亡的影响及其作用机制。方法 用细胞计数试剂(CCK-8)检测不同浓度五味子乙素对MDA-MB-231细胞存活率的影响;五味子乙素（10、20、40 μmol/L）作用 MDA-MB-231 细胞 24 h,分别用Annexin V-FITC/PI检测细胞凋亡情况;用DCFA-DA荧光探针检测细胞内活性氧（ROS）水平;用Western blot法检测细胞凋亡及内质网应激相关蛋白（Bcl-2、Bax、CHOP、GPR78、PERK、p-PERK、p-eIF2α、eIF2）的表达。结果 与空白组比较,随着五味子乙素浓度增大,细胞存活率明显降低,其IC₅₀为19.16 μmol/L;与对照组比较,五味子乙素（10、20、40 μmol/L）均能抑制细胞克隆形成(P<0.05),且呈剂量依赖;五味子乙素（10、20、40 μmol/L）均可诱导细胞凋亡（P<0.05）,使抗凋亡蛋白BCL-2的表达显著降低,促凋亡蛋白Bax的表达显著升高(P<0.05);五味子乙素（10、20、40 μmol/L）显著升高细胞内ROS水平(P<0.05),且呈剂量依赖;五味子乙素（10、20、40 μmol/L）能够激发内质网应激,使内质网应激相关蛋白CHOP、GPR78、p-eIF2α表达增多(P<0.05),且呈剂量依赖。结论 五味子乙素可能通过ROS介导内质网应激诱导MDA-MB-231细胞凋亡。     

基于Nrf2/HO-1信号通路研究橄榄苦苷的体外抗炎作用

目的 探讨橄榄苦苷对脂多糖（LPS）诱导的小鼠腹腔巨噬细胞（RAW264.7）炎症的保护作用及机制。方法 MTT法检测橄榄苦苷（0、10、20、40 μmol/L）对RAW264.7细胞活性的影响;用橄榄苦苷（10、20、40 μmol/L）预处理细胞1 h后,LPS诱导炎症模型,Griess试剂检测细胞内Nitrite释放;Western blotting方法检测细胞iNOS、COX-2、Nrf2、Keap-1、HO-1、NQO1蛋白表达水平;流式细胞仪检测细胞内NO、ROS、Ca²⁺的水平;荧光显微镜检测线粒体膜电位（MMP）水平;ELISA法检测细胞上清液中肿瘤坏死因子-α（TNF-α）、白介素-6（IL-6）的释放。结果 与模型组比较,橄榄苦苷组Nitrite释放水平显著降低（P<0.05、0.01、0.001）,iNOS、COX-2蛋白的表达显著降低（P<0.001）,NO的产生显著减少（P<0.05、0.01）;TNF-α、IL-6的释放受到显著抑制（P<0.001）;Ca²⁺释放受到显著抑制（P<0.01）;ROS生成受到显著抑制（P<0.01）;JC-1单体降低,恢复聚合物状态（红色荧光变多）,MMP稳定性增强;Keap1蛋白表达显著降低, Nrf2、HO-1、NQO1蛋白的表达均显著升高（P<0.05、0.01、0.001）。结论 橄榄苦苷对LPS诱导的RAW264.7细胞炎症具有抑制作用,其作用机制可能与激活Nrf2/HO-1信号通路相关。     

大黄素诱导人肝癌HepG2细胞线粒体凋亡作用研究

目的 研究大黄素对人肝癌HepG2细胞线粒体凋亡的影响。方法 培养人肝癌HepG2细胞,与5、10、20、40、60、80、100 μmol/L的大黄素作用24、48 h,MTS法检测细胞增殖;40、80、160 μmol/L大黄素作用HepG2细胞24 h,AO/EB双荧光染色法观察细胞凋亡的形态学改变;Annexin V/PI染色经流式细胞仪检测细胞凋亡;分光光度法检测caspase 3活性;ATP试剂盒检测细胞ATP含量,不同荧光探针加载后流式细胞仪测定大黄素对HepG2细胞内活性氧（ROS）含量、Ca²⁺浓度、线粒体膜电位（MMP）变化的影响。结果 大黄素抑制HepG2细胞生长,且呈时间、浓度相关性,半数抑制浓度（IC₅₀）为（77.42±1.25）μmol/L;随着大黄素浓度升高,AO/EB双染观察到细胞核浓缩、碎裂、凋亡小体等凋亡形态;与对照组比较,大黄素40、80、160 μmol/L作用于HepG2细胞24 h后细胞凋亡率显著增加,caspase 3活性显著增强,ROS水平、Ca²⁺浓度明显增加（P<0.05、0.01、0.001）,80、160 μmol/L组线粒体膜电位明显降低,ATP含量显著下降（P<0.05、0.01、0.001）。结论 大黄素造成HepG2细胞内ROS堆积,ATP合成功能障碍,线粒体膜电位明显下降,进而诱导线粒体通透转运孔开放,导致钙离子和细胞色素C外流,活化caspase蛋白家族,导致细胞凋亡。     

胡黄连苷I、II通过调节C/EBP-PPARγ通路抑制3T3-L1脂肪前体细胞的分化和脂肪合成

目的 探讨胡黄连主要有效成分胡黄连苷I、II对3T3-L1前脂肪细胞成脂分化的影响。方法 采用MTT法检测胡黄连苷I、II对3T3-L1细胞活力的影响,确定给药浓度;使用诱导分化培养基诱导3T3-L1细胞分化为成熟脂肪细胞,通过油红O染色法观察胡黄连苷I、II（20、40 μmol·L^-1）对细胞脂肪蓄积的影响;实时荧光定量PCR（qRT-PCR）法检测3T3-L1细胞脂肪合成相关的乙酰辅酶A羧化酶1（ACACA）、脂肪酸合酶（FASN）、硬脂酰辅酶A去饱和酶（SCD1）、脂肪酸结合蛋白（FABP4）和调节脂肪合成的转录因子过氧化物酶体增殖物激活受体γ（PPARγ）、固醇调节元件结合蛋白1（SREBP1）的mRNA表达水平;Western blotting实验检测CCAAT/增强子结合蛋白β（C/EBPβ）、SCD1和PPARγ的蛋白表达。结果 浓度低于50 μmol·L^-1的胡黄连苷I、II处理不会影响细胞的存活率;与对照组比较,模型组在诱导分化后细胞形态变圆,油红O染色显示细胞中有大量脂肪蓄积（P＜0.01）;与模型组比较,胡黄连苷I、II显著减少了脂肪蓄积（P＜0.01）。与对照组比较,模型组ACACA、FASN、SCD1、FABP4、PPARγ和SREBP1的mRNA表达均显著上调（P＜0.05、0.01）;与模型组比较,胡黄连苷I、II 20、40 μmol·L^-1均显著降低了ACACA、SCD1、FASN、FABP4、SREBP1 mRNA的表达（P＜0.05、0.01）,胡黄连苷I、II仅在40 μmol·L^-1时显著抑制SREBP1 mRNA的表达（P＜0.05、0.01）。Western blotting结果显示,与对照组比较,模型组PPARγ、C/EBPβ和SCD1的蛋白表达显著上调（P＜0.01）;与模型组比较,胡黄连苷I、II各浓度均显著降低了SCD1蛋白的表达（P＜0.01）,胡黄连苷I 40 μmol·L^-1和胡黄连苷II 20、40 μmol·L^-1组PPARγ、C/EBPβ蛋白的表达显著降低（P＜0.05、0.01）。结论 胡黄连苷I、II均可以显著抑制3T3-L1细胞的脂肪分化,并减少脂肪蓄积,胡黄连苷I表现出更好的剂量相关性,其作用机制可能与抑制C/EBPβ-PPARγ通路有关。     

槐耳颗粒联合DC-CIK细胞对乳腺癌MDA-MB-231干细胞体内外杀伤作用研究

目的 探究白屈菜红碱（CHE）对腺样囊性癌细胞（ACC2）生长的抑制作用及机制。方法 利用CCK8法、EdU法、Hoechst33342/PI双染色法、试剂盒法检测CHE对ACC2细胞活力、细胞增殖、细胞凋亡和活性氧（ROS）水平的影响;通过Western blotting技术检测CHE对Cleaved-Caspase 3、PARP、NF-κB、p-JNK、p-p38蛋白表达的影响;利用斑马鱼移植瘤模型检测CHE对斑马鱼体内ACC2细胞生长的抑制作用。结果 CCK-8结果显示：与对照组比较,2、3、4、5、6、7、8、9、10 μmol/L的CHE显著降低ACC2细胞的存活率（P<0.05、0.01）,且呈浓度相关性; ROS检测结果显示：与对照组比较,5、8 μmol/L的CHE导致ACC2细胞内的ROS水平显著上升（P<0.05、0.01）; EdU增殖检测结果表明：与对照组比较,5、8 μmol/L的CHE致使ACC2细胞的增殖能力显著下降（P<0.01）;Hoechst/PI染色结果显示：与对照组比较,CHE 5、8 μmol/L组ACC2细胞凋亡率显著上升（P<0.01）。抗氧化剂N-乙酰半胱氨酸（NAC）显著抑制CHE诱导的ROS水平升高、细胞凋亡增加（P<0.01）;Western blotting结果显示：2、5、8 μmol/L的CHE能够显著上调Cleaved-Caspase 3、PARP、NF-κB蛋白的表达（P<0.01）,且呈现浓度相关性,5、8 μmol/L的CHE能够显著上调p-JNK的蛋白表达（P<0.01）,8 μmol/L的CHE能够显著上调p-p38的蛋白表达（P<0.01）;NAC显著降低由CHE导致的Cleaved-Caspase 3、PARP、NF-κB、p-JNK、p-p38蛋白表达增加（P<0.01）,5、8 μmol/L CHE能够有效抑制斑马鱼体内肿瘤的生长（P<0.01）。结论 体外及斑马鱼移植瘤模型证明,CHE可以有效抑制ACC2细胞生长,其机制与提高细胞ROS水平,上调NF-κB、p-JNK、p-p38表达,从而抑制细胞增殖、诱导细胞凋亡相关。     

油酸诱导胰岛素抵抗HepG2细胞模型优化及小檗碱、黄芩苷、葛根素和甘草苷的体外降糖作用研究

目的 以油酸诱导脂代谢紊乱建立肝胰岛素抵抗（IR）细胞模型,研究中药单体成分小檗碱、黄芩苷、葛根素和甘草苷的体外降糖作用,为中药降糖组方或成分配伍优化提供研究基础。方法 采用油酸不同浓度（0.1、0.2、0.5、1.0 mmol/L）及不同作用时间点（24、36、48 h）诱导HepG2细胞,建立IR-HepG2细胞模型,葡萄糖氧化酶法测定细胞葡萄糖消耗量,CCK-8法检测细胞活力,确立模型的油酸最佳诱导浓度及最佳作用时间;蒽酮法和三酰甘油氧化酶法检测IR模型细胞内肝糖原和三酰甘油水平;油红O染色检测细胞形态的变化;观察葡萄糖消耗量检测IR模型的稳定性。采用油酸最佳诱导浓度及最佳作用时间建立IR-HepG2细胞模型,研究不同剂量小檗碱、葛根素、黄芩苷和甘草苷对细胞葡萄糖消耗量、糖原含量、三酰甘油及细胞活力的影响。结果 油酸诱导IR-HepG2细胞模型最佳浓度1 mmol/L,最佳作用时间24 h,此时与对照组比较,HepG2细胞肝糖原含量明显下降（P<0.001）,三酰甘油显著上升（P<0.01）,模型建立后可至少持续稳定36 h以上。与IR模型组比较,给药干预24 h后,不同浓度小檗碱（5、10、20、50 μmol/L）、黄芩苷（1、5、10、20、50 μmol/L）、葛根素（20、40、80、160 μmol/L）和1 μmol/L甘草苷显著增加葡萄糖消耗量（P<0.05、0.01、0.001）。与IR组比较,不同浓度小檗碱（10、20、50 μmol/L）、黄芩苷（20、50 μmol/L）、葛根素（10、20、80、160 μmol/L）显著提高肝糖原含量（P<0.001）;甘草苷升高糖原含量但是差异不显著。与对照组比较,除160 μmol/L葛根素和1 μmol/L黄芩苷显著抑制细胞活力（P<0.05）外,其他组对细胞活力的影响均不显著,结论 1 mmol/L油酸诱导24 h后能够建立稳定IR-HepG2细胞模型,适合作为高脂饮食诱导2型糖尿病的体外肝IR细胞模型。小檗碱、黄芩苷和葛根素显著增加油酸诱导的IR-HepG2细胞葡萄糖消耗量和糖原合成,进而改善脂代谢诱发的肝IR。     

芦荟大黄素对肝癌HepG2细胞放射敏感性的影响

目的 探讨芦荟大黄素（AE）对肝癌HepG2细胞放射敏感性的影响。方法 采用MTT法测定AE对HepG2细胞的IC₅₀值,作为后续实验的药物浓度。将对数生长期HepG2细胞分为对照组、AE组、γ射线组及联合组,对照组给予常规培养液处理,AE组给予IC₅₀浓度的AE处理,γ射线组给予8 Gy γ射线处理,联合组给予IC₅₀浓度的AE联合8 Gy γ射线处理,每组细胞均培养48 h。采用MTT法检测细胞增殖抑制率,微核试验检测细胞DNA损伤情况,流式细胞术检测细胞凋亡,克隆形成实验观察AE对HepG2细胞的放射增敏作用,Western blotting检测p53及γ-H2AX蛋白表达水平。结果 AE对HepG2细胞的IC₅₀值为60 μmol·L^-1,以此作为后续实验的药物浓度。与对照组比较,AE组、γ射线组及联合组细胞增殖抑制率、微核率、凋亡率及p53、γ-H2AX蛋白表达水平均显著升高（P<0.05）,且联合组升高趋势最为明显（P<0.05）;与γ射线组比较,联合组D₀、Dq及SF显著降低（P<0.05）,SER为（1.81±0.24）。结论 AE和γ射线均对HepG2细胞的增殖有抑制作用,并且AE对HepG2细胞具有放射增敏作用。     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Lung disease and PKCs

The lung offers a rich opportunity for development of therapeutic strategies focused on isozymes of protein kinase C (PKCs). PKCs are important in many cellular responses in the lung, and existing therapies for pulmonary disorders are inadequate. The lung poses unique challenges as it interfaces with air and blood, contains a pulmonary and systemic circulation, and consists of many cell types. Key structures are bronchial and pulmonary vessels, branching airways, and distal air sacs defined by alveolar walls containing capillaries and interstitial space. The cellular composition of each vessel, airway, and alveolar wall is heterogeneous. Injurious environmental stimuli signal through PKCs and cause a variety of disorders. Edema formation and pulmonary hypertension (PHTN) result from derangements in endothelial, smooth muscle (SM), and/or adventitial fibroblast cell phenotype. Asthma, chronic obstructive pulmonary disease (COPD), and lung cancer are characterized by distinctive pathological changes in airway epithelial, SM, and mucous-generating cells. Acute and chronic pneumonitis and fibrosis occur in the alveolar space and interstitium with type 2 pneumocytes and interstitial fibroblasts/myofibroblasts playing a prominent role. At each site, inflammatory, immune, and vascular progenitor cells contribute to the injury and repair process. Many strategies have been used to investigate PKCs in lung injury. Isolated organ preparations and whole animal studies are powerful approaches especially when genetically engineered mice are used. More analysis of PKC isozymes in normal and diseased human lung tissue and cells is needed to complement this work. Since opposing or counter-regulatory effects of selected PKCs in the same cell or tissue have been found, it may be desirable to target more than one PKC isozyme and potentially in different directions. Because multiple signaling pathways contribute to the key cellular responses important in lung biology, therapeutic strategies targeting PKCs may be more effective if combined with inhibitors of other pathways for additive or synergistic effect. Mechanisms that regulate PKC activity, including phosphorylation and interaction with isozyme-specific binding proteins, are also potential therapeutic targets. Key isotypes of PKC involved in lung pathophysiology are summarized and current and evolving therapeutic approaches to target them are identified.     

Gender-related symptoms and correlates of alcohol dependence among men and women with a lifetime diagnosis of alcohol use disorders

This study explored gender-related symptoms and correlates of alcohol dependence in a crosssectional study of 150 men and 150 women with a lifetime diagnosis of alcohol use disorders (AUD). Participants were recruited in equal numbers from treatment settings, correctional centres and the general community. Standardized measures were used to determine participants' use of substances, history of psychiatric disorders and psychosocial stress, their sensation seeking and family history of substance use and mental health disorders. Multivariate analyses were used to detect patterns of variables associated with gender and the lifetime severity of AUD. Men had a longer history of severe AUD than women. Women had similar levels of alcohol dependence and medical and psychological sequelae as men, despite 6 fewer years of AUD. More women than men had a history of severe psychosocial stress, severe dependence on other substances and antecedent mental health problems, especially mood and anxiety disorders. There were differences in family history of alcohol-related problems approximating same-gender aggregation. The severity of a lifetime AUD was predicted by its earlier age at onset and the occurrence of other disorders, especially anxiety, among both men and women. The limitations in the generalizability of these findings due to sample idiosyncrasies are discussed.     

Biochemical and molecular characterisation of cubozoan protein toxins

Class Cubozoa includes several species of box jellyfish that are harmful to humans. The venoms of box jellyfish are stored and discharged by nematocysts and contain a variety of bioactive proteins that are cytolytic, cytotoxic, inflammatory or lethal. Although cubozoan venoms generally share similar biological activities, the diverse range and severity of effects caused by different species indicate that their venoms vary in protein composition, activity and potency. To date, few individual venom proteins have been thoroughly characterised, however, accumulating evidence suggests that cubozoan jellyfish produce at least one group of homologous bioactive proteins that are labile, basic, haemolytic and similar in molecular mass (42-46 kDa). The novel box jellyfish toxins are also potentially lethal and the cause of cutaneous pain, inflammation and necrosis, similar to that observed in envenomed humans. Secondary structure analysis and remote protein homology predictions suggest that the box jellyfish toxins may act as α-pore-forming toxins. However, more research is required to elucidate their structures and investigate their mechanism(s) of action. The biological, biochemical and molecular characteristics of cubozoan venoms and their bioactive protein components are reviewed, with particular focus on cubozoan cytolysins and the newly emerging family of box jellyfish toxins.     

Dose tolerability of chronically inhaled voriconazole solution in rodents

Invasive pulmonary aspergillosis (IPA) is a fungal disease of the lung associated with high mortality rates in immunosuppressed patients despite treatment. Targeted drug delivery of aqueous voriconazole solutions has been shown in previous studies to produce high tissue and plasma drug concentrations as well as improved survival in a murine model of IPA. In the present study, rats were exposed to 20 min nebulizations of normal saline (control group) or aerosolized aqueous solutions of voriconazole at 15.625 mg (low dose group) or 31.25 mg (high dose group). Peak voriconazole concentrations in rat lung tissue and plasma after 3 days of twice daily dosing in the high dose group were 0.85 ± 0.63 μg/g wet lung weight and 0.58 ± 0.30 μg/mL, with low dose group lung and plasma concentrations of 0.38 ± 0.01 μg/g wet lung weight and 0.09 ± 0.06 μg/mL, respectively. Trough plasma concentrations were low but demonstrated some drug accumulation over 21 days of inhaled voriconazole administered twice daily. Following multiple inhaled doses, statistically significant but clinically irrelevant abnormalities in laboratory values were observed. Histopathology also revealed an increase in the number of alveolar macrophages but without inflammation or ulceration of the airway, interstitial changes, or edema. Inhaled voriconazole was well tolerated in a rat model of drug inhalation.