HMS-01在小鼠体内的药动学研究

目的 研究HMS-01在小鼠体内的药动学,为后续研究、提供支持。方法 采用液相色谱-串联质谱(LC-MS/MS)技术,建立灵敏、特异的测定血浆等生物样品中HMS-01浓度的分析方法,用建立的方法开展HMS-01在C57BL/6J小鼠体内药动学研究。分别对其进行了3个剂量单次灌胃给药、1个剂量单次静注给药的药动学研究,以获得基本药动学参数。结果 小鼠药代动力学结果表明,HMS-01肠道吸收快,小鼠口服生物利用度中等(50%～70%)。HMS-01在小鼠体内的暴露水平(AUC和c_max)随剂量的增加而增加,其中AUC随剂量的增加成线性相关。HMS-01静脉给药后,在小鼠体内的半衰期不长(约1 h);血浆清除率(CL_tot,p)为2.8L/(h·kg),与小鼠肝血流量相当;表观分布容积(V_SS)为5 L/kg,远大于小鼠总体液。雌雄小鼠经HMS-01口服30、60mg/kg,在AUC和F有显著差异(P<0.05),在c_max、AUC_0-∞、t_1/2、C     

HMS-01在大鼠体内的药动学研究

目的研究HMS-01在大鼠体内的药动学,为后续研究提供支持。方法采用液相色谱-串联质谱(LCMS/MS)技术,建立灵敏、特异的测定血浆等生物样品中HMS-01浓度的分析方法,用建立的方法开展HMS-01在大鼠体内药动学研究。在SD大鼠上分别进行了1个剂量单次灌胃给药、1个剂量单次静注给药的药动学研究,以获得基本药动学参数。结果大鼠静脉注射1 mg/kg的HMS-01后,雄性与雌性大鼠血浆浓度-时间曲线下面积(AUC0-t)分别为221和409 ng·h/ml,平均清除率分别为4.53和2.41 L/h·kg,平均血浆消除半衰期分别为0.786和1.27 h,表观分布容积分别为5.13和3.82 L/kg。灌胃给予30 mg/kg的HMS-01后,在大鼠体内血浆浓度达峰时间tmax为1.17 h,达峰浓度cmax为1243 ng/ml,消除半衰期(t1/2)为2.00 h。雄、雌大鼠AUC0-t分别为2271和8529 ng·h/ml,生物利用度分别为34.3%和69.5%。结论HMS-01在大鼠体内的药动学过程存在显著的性别差异,口服吸收较好,雌性大鼠的生物利用度远高于雄性。     

手动膜片钳检测盐酸罗哌卡因及其右旋异构体对HEK293细胞hERG电流的影响

目的 研究比较盐酸罗哌卡因和盐酸罗哌卡因右旋异构体对高表达hERG钾通道的HEK293细胞hERG电流的影响。方法 用手动膜片钳检测转染后hERG钾通道稳定表达的HEK293细胞电流,多菲莱德做阳性药,将盐酸罗哌卡因和盐酸罗哌卡因右旋异构体依次稀释成30.00、10.00、3.33、1.11、0.37μmol·L^-1,依次作用于细胞,记录电流变化,计算抑制率。结果 盐酸罗哌卡因0.37、1.11、3.33、10、30μmol·L^-1对电流Iherg-tail的抑制率分别为(6.12±0.30)%、(13.04±1.20)%、(19.21±0.33)%、(35.56±0.66)%、(65.37±4.17)%,IC₅₀为19.482μmol·L^-1(n=15)。盐酸罗哌卡因右旋异构体0.37、1.11、3.33、10.00、30.00μmol·L^-1对电流Iherg-tail的抑制率分别为(4.13±3.43)%、(7.34±5.60)%、(9.49±2.75)%、(16.60±0...     

探讨蛋白酶体抑制剂抑制晶状体上皮细胞增殖的机制

目的 探讨蛋白酶体抑制剂MG132抑制晶状体上皮细胞系SRA01/04增殖的机制.方法 MTT比色法检测MG132对SRA01/04细胞增殖的影响,流式细胞术检测MG132对SRA01/04细胞凋亡的影响,荧光显微镜下Annexin V/FITC-PI双染法观察MG132对SRA01/04细胞的影响.结果 MG132随着浓度的增高对SRA01/04细胞增殖的抑制作用逐渐增强,作用36 h时半数有效抑制浓度IC50为2.48μmol/L.2.5、5.0μmol/L MG132分别与SRA01/04细胞作用24 h,早期细胞凋亡率分别为（5.75±0.24）％和（10.23±2.68）％,而对照组为（0.34±0.26）％,差异有统计学意义（P＜0.05）.结论 MG132诱导细胞凋亡来抑制SRA01/04细胞的增殖,可起到防治后发性白内障的作用.     

妊娠糖尿病患者胰岛β细胞功能和胰岛素抵抗的评价及临床意义

目的 探讨妊娠糖尿病患者胰岛B细胞功能和胰岛素抵抗及临床意义。方法 孕25—32周经初筛和OGTT（同时做胰岛素释放试验）确诊的妊娠糖尿病患者56例，并于分娩后6—8周复查OG—TT（同时做胰岛素释放试验），其中31例为葡萄糖耐量正常（NGT），分娩前后样本分别确定为GDM1和NGT；25例葡萄糖耐量受损（IGT），分娩前后样本分别定为样本GDM2和IGT，四份样本均考查胰岛素曲线下面积（INS ACC）、β细胞功能指数（MBCI）及Homa胰岛素抵抗指数（Homa—IR）的变化。血浆葡萄糖、胰岛素分别采用葡萄糖氧化酶法和放免分析法测定。结果 GDM2、GDM1、IGT、NGT的血糖值依次降低，四份两两比较，除GDM2和GDM1外差异均存在统计学意义（P〈0．001）。GDM2、GDM1、IGT、NGT的INS。。值依次降低，四份两两比较，差异均存在统计学意义（P〈0．01）。NGT、IGT、GDM1、GDM2的MBCI值依次降低，四份两两比较，差异有统计学意义（P〈0．01）。GDM2、GDM1、IGT、NGT的Homa—IR值依次降低，四份两两比较，差异均存在统计学意义（P〈0．01）。结论 妊娠糖尿病患者胰岛β细胞功能越差，胰岛素抵抗越强，分娩后患IGT的可能性越大。     

红景天苷衍生物S01对谷氨酸及缺氧缺糖所致SH-SY5Y细胞损伤的保护作用

目的探讨红景天苷衍生物S01(3′,4′,5′-三甲氧基苄基-1-硫代-β-D-吡喃葡萄糖苷)对神经细胞的保护作用,并初步探讨其作用机制。方法用不同浓度S01(2,10和50mg.L-1)预处理SH-SY5Y细胞,12h后用谷氨酸或连二亚硫酸钠(Na2S2O4)诱导SH-SY5Y细胞损伤,用MTT法检测细胞存活率,比色法检测乳酸脱氢酶(LDH)释放、超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量,荧光探针负载法检测细胞内活性氧(ROS)水平和细胞内游离钙离子浓度([Ca2+]i),化学发光法检测细胞中腺苷三磷酸(ATP)水平。结果在谷氨酸所致SH-SY5Y细胞损伤模型中,与对照组比较,模型组细胞存活率、SOD活性和ATP水平降低,LDH释放、MDA含量和细胞内ROS水平增加。与模型组比较,S01(2,10和50mg.L-1)可提高细胞存活率,减少谷氨酸引起的LDH释放,拮抗谷氨酸引起的SOD活性和ATP水平降低,并拮抗MDA和ROS水平升高。在Na2S2O4所致SH-SY5Y细胞缺氧缺糖损伤模型中,与对照组比较,模型组细胞存活率下降,LDH释放增多,细胞[Ca2+]i升高。与模型组比较,S01(2,10和50mg.L-1)可提高细胞存活率,减少缺氧缺糖引起的LDH释放和细胞内钙超载。结论S01对谷氨酸和缺氧缺糖所致神经细胞损伤具有保护作用,提示S01可能对脑缺血具有治疗作用。     

紫衫醇脂质体对卵巢癌细胞增殖及转移能力的影响及其相关机制

目的研究紫杉醇脂质体对卵巢癌细胞增殖及转移能力的影响及其相关机制。方法分别用2. 5μg·m L^-1紫杉醇脂质体及等量0. 9%Na Cl处理人卵巢癌细胞SKOV-3 24 h,作为本研究的实验组和对照组。用噻唑蓝染色法检测2组细胞的增殖能力,用Transwell侵袭及迁移实验检测2组细胞的侵袭迁移能力,用荧光定量链式聚合酶反应技术检测2组细胞微小RNA-499(miR-499)和生长抑制特异性转录本5(GAS5)的表达水平,用Western Blot法检测2组细胞骨髓细胞瘤病毒癌基因同源物(c-Myc)分子的表达水平。结果实验组和对照组的增殖活力分别为(0. 45±0. 01)和(0. 75±0. 03) OD,穿透Transwell基底膜的细胞数分别为(44. 9±7. 5)和(83. 6±11. 7)个,迁出Transwell小室孔的细胞数分别为(67. 0±10. 2)和(91. 4±12. 1)个,miR-499相对表达水平分别为(0. 09±0. 01)和(0. 21±0. 01),GAS5相对表达水平分别为(0. 14±0. 01)和(0. 06±0. 01),c-Myc分子的表达水平分别为(0. 32±0. 05)和(0. 65±0. 09)DPI,差异均有统计学意义(均P <0. 05)。结论紫杉醇脂质体可有效抑制卵巢癌细胞增殖及转移能力,其机制与调控miR-499-GAS5-c-Myc表达水平有关。     

草酸铂对大鼠神经节细胞元细胞膜上钠离子通道电流影响的实验研究

目的 初步观察草酸铂(抗肿瘤药)神经毒性的作用机制.方法 分离大鼠脊髓背根神经节神经元细胞,用膜片钳法分别检测草酸铂细胞外和细胞内给药时,神经元细胞膜七钠离子通道电流的变化.结果 在给药5、10、15 min,与给药前比较,钠离子电流振幅大、中、小细胞都有明显降低(P<0.05);小细胞给药后不同时间点的钠离子电流振幅抑制率明显大于同期大、中细胞的抑制率,其对小细胞的抑制作用明显高于大细胞和中细胞(P<0.05).结论 草酸铂对神经元细胞膜上电压门控钠离子通道的抑制可能是其神经毒性作用机制;背根神经节小细胞可能与草酸铂神经毒性症状的产生关系较密切.     

寡核苷酸促人牙周膜细胞增殖及成骨分化的实验研究

目的研究寡核苷酸(ODN)mt01对人牙周膜细胞增殖和向成骨细胞分化的影响,为临床药剂的开发奠定实验基础。方法采用人离体牙牙周膜细胞作为研究对象,原代培养牙周膜细胞,确定细胞最佳铺板浓度后,通过四甲基偶氮唑盐(MTT)比色法研究ODN mt01对人牙周膜细胞增殖的影响;用碱性磷酸酶(ALP)试剂盒检测ODN mt01影响人牙周膜细胞成骨分化的最佳工作浓度。结果分别作用于人牙周膜细胞24、48、72和96 h,ODN mt01在整个作用过程中均表现出显著的促人牙周膜细胞增殖作用(P<0.05);与磷酸盐缓冲液(PBS)对照组ALP表达量相比,工作浓度为1.0 mg/L时,ODN mt01可促进人牙周膜细胞表达ALP水平显著升高(P<0.05)。结论适当工作浓度的ODN mt01可以促进体外培养的人牙周膜细胞增殖和成骨分化。     

ERK抑制剂U0126通过下调cyclin D1与survivin蛋白表达抑制乳腺癌细胞增殖

目的研究ERK通路抑制剂U0126对乳腺癌细胞增殖的抑制作用,并探讨其调控机制。方法培养人乳腺癌细胞株MCF-7、MDA-MB231,将细胞分组干预,MTT法检测各组细胞增殖情况;流式细胞仪检测各组细胞周期及细胞凋亡; Western blot检测细胞中p-ERK/ERK、cyclin D1、survivin及cleaved caspase-3蛋白表达。结果MTT结果显示,U0126干预24、48 h后,MCF-7、MDA-MB231细胞抑制率明显增加(P <0. 01); MCF-7、MDA-MB231细胞经U0126干预24 h后,检测细胞周期及细胞凋亡,G_0/G_1期细胞比例较对照组明显增加,S及G_2期细胞比例下降(P <0. 05);而干预组细胞凋亡率较未干预组明显增多(P <0. 01); U0126处理人乳腺癌细胞2 h后,可阻断ERK的磷酸化,减少cyclin D1的蛋白表达,抑制survivin而上调cleaved caspase-3的表达,与对照组相比差异有显著性(P <0. 01)。结论 U0126通过阻断MCF-7、MDA-MB231细胞中ERK信号通路,调控cyclin D1,将细胞周期阻断在G_0/G_1期,抑制survivin而增加cleaved caspase-3表达,增加细胞凋亡,继而使乳腺癌细胞MDA-MB231、MCF-7的增殖受到抑制。     

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.