津力达颗粒对高糖作用下小鼠胰岛β细胞增殖和凋亡的影响

目的 探讨津力达颗粒对高糖作用下小鼠胰岛β细胞增殖和凋亡的影响。方法 将MIN6细胞分为正常对照(NC)组、津力达(JLD)组、高糖(HG)组和高糖+津力达(HG+JLD)组。培养48 h后,分别用2.8,16.7 mmol·L^-1葡萄糖KRBH缓冲培养液孵育0.5 h,酶联免疫吸附实验(ELISA)检测上清液胰岛素含量。采用噻唑蓝(MTT)法检测细胞增殖率,流式细胞仪检测细胞凋亡率,免疫印迹法(Western blotting)检测Caspase-3、Bax、Bcl-2和Smad2/3蛋白表达。结果 与2.8 mmol·L^-1葡萄糖刺激比较,16.7 mmol·L^-1葡萄糖刺激时各组胰岛素分泌(GSIS)均升高(均P<0.01)。16.7 mmol·L^-1葡萄糖刺激时HG+JLD组GSIS高于HG组(P<0.01)。与HG组比较,HG+JLD组细胞增殖率增高(P<0.01),早晚期凋亡率降低(均P<0.01),Caspase-3-2、Bax蛋白表达下调(P<0.01或...     

新绿原酸抑制血管生成的研究

目的 考察新绿原酸(5-caffeoylquinic acid, 5-CQA)对血管生成的影响。方法 应用人脐内皮细胞(HUVEC)、大鼠动脉环模型和转基因血管荧光斑马鱼模型考察5-CQA对血管生成的抑制作用。同时建立肿瘤细胞斑马鱼移植瘤模型，考察5-CQA对肠下静脉(subintestinal vein, SIV)生成的作用。结果 5-CQA在320μmol·L^-1及以下浓度对HUVEC的增殖无明显影响，5-CQA能够抑制HUVEC体外迁移及细胞中VEGF、bFGF mRNA的表达；与对照组比较，56和112μmol·L^-1的5-CQA具有显著抑制大鼠动脉环微血管生成的作用(P<0.05);在斑马鱼血管荧光模型中56和112μmol·L^-1的5-CQA具有显著抑制24 h斑马鱼节间血管(intersegmental vessel, ISV)生成的作用；同时5-CQA对斑马鱼移植瘤的肠下静脉生成具有明显抑制作用(P<0.05)。结论 5-CQA具有抑制血管生成的作用，其作用机制与抑制VEGFR2受体的表达有关...     

天花粉对人乳腺癌MCF-7细胞凋亡及相关基因表达的影响

目的研究天花粉对人乳腺癌MCF-7细胞凋亡及相关基因表达的影响。方法取对数生长期人乳腺癌MCF-7细胞以30,60,90,120μg·m L^-1的天花粉蛋白处理,作为实验组,对照组用等量的0.9%Na Cl处理,继续培养24,48,72h。观察并比较各组细胞形态学及细胞核凋亡形态学变化,以噻唑蓝(MTT)比色法检测各组人乳腺癌MCF-7细胞增殖抑制情况,分光光度法检测天花粉蛋白对人乳腺癌MCF-7细胞胱天蛋白酶(caspase)-3和caspase-8表达的影响。结果实验组人乳腺癌MCF-7细胞呈现明显的细胞凋亡及核凋亡现象,且随天花粉蛋白质量浓度的增加,其细胞核凋亡现象越明显;干预24 h后,30,60,90,120μg·mL^-1实验组增殖抑制率分别为(1.61±0.94)%,(2.81±1.01)%,(7.05±1.03)%和(9.59±1.12)%;干预48 h后增殖抑制率分别为(2.13±1.01)%,(6.14±1.12)%,(9.05±1.09)%和(19.60±1.15)%;干预72 h后增殖抑制率分别为(3.28±1.07)%,(7.18±1.51)%,(18.39±1.81)%和(29.59±1.63)%。实验组人乳腺癌MCF-7细胞增殖抑制率随作用时间的延长及天花粉蛋白质量浓度的增大而逐渐升高(均P<0.01);90μg·m L^-1实验组24,48,72 h caspase-3分别为1.49±0.15,2.36±0.25,2.15±0.23;caspase-8分别为1.94±0.26,1.56±0.19,1.39±0.20。对照组人乳腺癌MCF-7细胞caspase-3及caspase-8随作用时间的延长无显著变化(均P>0.05),而90μg·mL^-1实验组caspase-3先升高后降低(P<0.05或P<0.01),以48 h时最高;caspase-8则逐渐降低(P<0.01);且各时间点90μg·m L^-1实验组caspase-3及caspase-8均显著高于对照组(均P<0.01)。结论天花粉蛋白可有效促进人乳腺癌MCF-7细胞凋亡,抑制其增殖,且增殖抑制作用呈时间-剂量依赖性,其作用机制可能与上调人乳腺癌MCF-7细胞caspase-3及caspase-8表达相关。     

利拉鲁肽抗CXC趋化因子配体16诱导的人足细胞脂质沉积作用及机制

目的 探讨利拉鲁肽(liraglutide)抗CXC趋化因子配体16(CXCL16)诱导的人足细胞脂质沉积的作用及机制。方法 (1)葡萄糖40 mmol·L^-1刺激人足细胞24 h，棕榈酸250μmol·L^-1刺激人足细胞6 h,Western印迹法检测足细胞CXCL16蛋白表达水平。(2)重组CXCL16 100μg·L^-1刺激足细胞0,6,12和24 h,Western印迹法检测足细胞裂隙素蛋白表达水平。(3)足细胞分为细胞对照组、CXCL16 100μg·L^-1组、CXCL16+利拉鲁肽10,50和100 nmol·L^-1组及CXCL16+辛伐他汀100 nmol·L^-1组，加药2 h后加重组CXCL16 100μg·L^-1继续培养24 h，油红O染色检测足细胞脂滴面积。(4)足细胞分为细胞对照组、CXCL16100μg·L^-1组、CXCL16+利拉鲁肽100 nmol·L^-1组和CX...     

地西他滨联合丙戊酸钠对白血病细胞株HL-60促凋亡和分化的影响

目的探讨地西他滨（DCA）和丙戊酸钠（VPA）联用对白血病细胞株HL-60促凋亡和分化诱导的协同作用。方法白血病细胞株HL-60分别与以下药物终浓度组作用48 h:对照组,DCA单药A组(1.0μmol·L^-1),DCA单药B组(4.0μmol·L^-1),VPA单药组(2.0 mmol·L^-1),联合用药A组(IDCA 1.0μmol·L^-1+VPA 2.0 mmol·L^-1),联合用药B组(DCA 4.0μmol·L^-1+VPA2.0 mmol·L^-1)。应用Annexin V-FTTC/PI标记法检测早期凋亡率,流式细胞术检测CD34、CD117平均荧光强度（MFI）以及CD11b、CD14表达率。结果联合用药A、B组的凋亡率高于其各自的单药组,差异具有高度统计学意义（P<0.01）;联合用药A、B组的CD117和CD34 MFI低于其各自的单药组,差异具有高度统计学意义（P<0.01）;联合用药A组的CD11b和CD14表达率以及联合用药B组的CD11b表达率均低于其各自的单药组,差异具有高度统计学意义（P<0.01）。结论白血病细胞株HL-60中VPA能显著增强DCA的促凋亡分化作用。     

参芪复方对高糖“代谢记忆”介导的人脐静脉内皮细胞的影响

目的 初步探讨参芪复方对高糖“代谢记忆”介导的人脐静脉内皮细胞（HUVEC）的影响。方法 将HUVEC在高糖(30 mmol·L^-1)中培养3 d,再转入正常糖(5.5 mmol·L^-1)培养4 d,建立高糖“代谢记忆”细胞模型。将造模成功的细胞分为对照组和低、中、高剂量实验组;另取正常细胞作为空白组和模型组。对照组给予25 mmol·L^-1白藜芦醇干预3 d;低、中、高剂量实验组分别给予100,200和400μL·mL^-1参芪复方含药血清干预3 d;空白组给予5.5 mmol·L^-1葡萄糖干预3 d;模型组给予30.0 mmol·L^-1葡萄糖干预3 d。用CCK-8法检测细胞增殖活性,用酶联免疫吸附实验法检测细胞白细胞介素-6（IL-6）水平。结果 低、高剂量实验组和对照组、空白组、模型组在108 h时细胞活性（光密度值）分别为0.78±0.05,1.41±0.14,1.92±0.36,1.39±0.01和0.64±0.02,IL-6水平分别为21.29...     

亚精胺对脂多糖诱导损伤的人脐静脉内皮细胞自噬的影响及机制

目的 探讨亚精胺(SPD)对血管内皮细胞损伤后自噬的影响及机制。方法 (1) SPD 50,100 和200 μmol·L^-1预处理人脐静脉内皮细胞(HUVEC)3 h,加脂多糖(LPS)1 mg·L^-1孵育 24 h,CCK-8 检测细胞存活率。(2) HUVEC 细胞分成 7 组,细胞对照组、模型组、模型+SPD(100 μmol·L^-1预处理 3 h)组、模型+腺苷酸激活蛋白激酶(AMPK)抑制剂多索霉素(Dor,10 μmol·L^-1预处理 1 h)组、模型+SPD+Dor 组、模型+AMPK激活剂阿卡地新(Aicar,0.5 mmol·L^-1预处理1.5 h)组和模型+SPD+Aicar组,药物预处理结束,模型组和用药组加 LPS 1 mg·L^-1共孵育 24 h。流式细胞术检测细胞凋亡,免疫荧光和 Western印迹法检测微管相关蛋白 1 轻链 3-Ⅰ(LC3-Ⅰ)、LC3-Ⅱ和 P62 蛋白表达水平,Western 印迹法检测细胞 AMPK/...     

归芍方对邻苯二甲酸二丁酯诱导的斑马鱼眼部细胞凋亡的影响

目的 探讨归芍方对斑马鱼眼部细胞凋亡模型的影响及其不同提取方法与药理作用间关系,进一步研究归芍方抗眼部细胞凋亡作用相关机制。方法 将受精后30 h斑马鱼分为正常对照组、归芍方药材水提取物(ME)组、水提醇沉提取物(MEA)组和提取物配方(PS)组,于给药后24和48 h观察斑马鱼存活率。吖啶橙染色观察药物对斑马鱼眼部细胞凋亡的影响。将受精后30 h斑马鱼分为正常对照组、模型对照组、人参皂苷Rg1组和低中高浓度ME、MEA、PS组。模型对照组给予10μmol·L^-1邻苯二甲酸二丁酯(DBP),人参皂苷Rg1组及归芍方各给药组于造模同时分别给予1μmol·L^-1人参皂苷Rg1和10,50,100μg·mL^-1ME、MEA及PS,培养18 h后观察眼部细胞凋亡数目。免疫印迹(Western blotting)法检测凋亡相关蛋白Bcl-2表达,实时荧光定量聚合酶链反应(PCR)检测线粒体mtDNA拷贝数变化。结果 归芍方各提取物100μg·mL^-1及其以下浓度对斑马鱼存活率无显著影响;1μmol·L     

氧化苦参碱对SGC-7901胃癌细胞增殖及对血管内皮生长因子表达的影响

目的研究中药苦参有效成分氧化苦参碱对SGC-7901胃癌细胞增殖及对血管内皮生长因子(VEGF)表达的影响。方法用质量浓度为0.5,1.0,2.0,4.0,8.0 g·L^-1的氧化苦参碱干预SGC-7901胃癌细胞分别为0.5,1.0,2.0,4.0,8.0 g·L^-1质量浓度实验组,无任何药物处理的SGC-7901胃癌细胞为对照组。用噻唑蓝(MTT)法检测各组SGC-7901胃癌细胞培养第1,2,3,4天的增殖情况。用1.0,2.0和4.0 g·L^-1氧化苦参碱处理SGC-7901细胞并培养48 h后,用实时荧光定量-聚合酶链式反应(RT-PCR)法检测各组VEGF mRNA的表达情况。用酶联免疫吸附(ELISA)法检测各组蛋白表达情况。结果细胞培养第4天,对照组和0.5,1.0,2.0,4.0,8.0 g·L^-1实验组的增殖抑制率分别为(4.15±3.93)%,(14.79±1.66)%,(31.65±2.07)%,(41.57±2.93)%,(64.37±4.13)%,(79.17±4.75)%,与对照组相比,各质量浓度实验组SGC-7901胃癌细胞增殖抑制率均显著升高(P<0.01)。实验组的增殖抑制率随着培养时间和药物浓度的增加而增加(P<0.05或P<0.01)。氧化苦参碱干预48 h后,对照组和1.0,2.0和4.0 g·L^-1实验组VEGF mRNA表达量分别为0.82±0.03,0.65±0.05,0.54±0.06,0.46±0.04,与对照组比较,实验组VEGF mRNA表达量明显降低,且随氧化苦参碱质量浓度的升高其VEGF mRNA表达量呈逐渐降低趋势,组间差异均有统计学意义(均P<0.01)。对照组和0.5,1.0,2.0,4.0,8.0 g·L^-1实验组上清液中VEGF蛋白表达量分别为(1326.52±139.57),(1305.69±119.88),(1235.59±102.36),(1083.33±89.65),(789.85±95.26),(426.59±83.16)pg·mL^-1,与对照组相比,2.0,4.0,8.0 g·L^-1实验组细胞上清中VEGF蛋白表达量随氧化苦参碱质量浓度的增加而逐渐降低(P<0.05)。结论氧化苦参碱可抑制SGC-7901胃癌细胞增殖,增殖抑制率与作用时间、药物浓度呈正比,同时氧化苦参碱具有抑制相关肿瘤血管生成因子表达的作用。     

小剂量氯胺酮联用丁基苯酞的抗抑郁作用及其机制

目的 研究小剂量氯胺酮(Ket)联用丁基苯酞(NBP)的抗抑郁作用及其机制。方法 建立皮质酮(CORT)诱导PC12细胞损伤的体外抑郁模型,PC12细胞分成6组：空白对照组、模型对照组(CORT 400μmol·L^-1)、对照A组(CORT 400μmol·L^-1+NBP 1μmol·L^-1)、对照B组(CORT 400μmol·L^-1+Ket 0.01μmol·L^-1)、阳性对照组(CORT 400μmol·L^-1+Ket 0.1μmol·L^-1)、联合组(CORT 400μmol·L^-1+Ket 0.01μmol·L^-1+NBP 1μmol·L^-1)。CCK-8法检测细胞存活率,荧光显微镜观察细胞神经元形态变化,蛋白免疫印迹法评价联合用药对损伤细胞p-ERK/ERK、脑源性神经营养因子(BDNF)、synapsin蛋白表达变化。将ICR小鼠随机分为模型对照组、NBP...     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

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.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.     

Aquatic Insects and Trace Metals: Bioavailability,Bioaccumulation, and Toxicity

Abstract

The uptake of metals from food and water sources by insects is thought to be additive. For a given metal, the proportions taken up from water and food will depend both on the bioavailable concentration of the metal associated with each source and the mechanism and rate by which the metal enters the insect. Attempts to correlate insect trace metal concentrations with the trophic level of insects should be made with a knowledge of the feeding relationships of the individual taxa concerned. Pathways for the uptake of essential metals, such as copper and zinc, exist at the cellular level, and other nonessential metals, such as cadmium, also appear to enter via these routes. Within cells, trace metals can be bound to proteins or stored in granules. The internal distribution of metals among body tissues is very heterogeneous, and distribution patterns tend to be both metal and taxon specific. Trace metals associated with insects can be both bound on the surface of their chitinous exoskeleton and incorporated into body tissues. The quantities of trace meals accumulated by an individual reflect the net balance between the rate of metal influx from both dissolved and particulate sources and the rate of metal efflux from the organism. The toxicity of metals has been demonstrated at all levels of biological organization: cell, tissue, individual, population, and community. Much of the literature pertaining to the toxic effects of metals on aquatic insects is based on laboratory observations and, as such, it is difficult to extrapolate the data to insects in nature. The few experimental studies in nature suggest that trace metal contaminants can affect both the distribution and the abundance of aquatic insects. Insects have a largely unexploited potential as biomonitors of metal contamination in nature. A better understanding of the physico-chemical and biological mechanisms mediating trace metal bioavailability and exchange will facilitate the development of general predictive models relating trace metal concentrations in insects to those in their environment. Such models will facilitate the use of insects as contaminant biomonitors.