Study of the mechanism on the apoptosis induced in Human leukemia cell line K562 by the combination of indole-3-acetic acid and horseradish peroxidase

Objective To investigate the mechanisms of apoptosis induced in Human leukemia cell line K562 by the combination of indole-3-acetic acid and horseradish peroxidase. Methods Human leukemia cell line K562 were exposed to indole-3-acetic acid （IAA） at 20, 40, 60, 80 or 100 mol/L and horseradish peroxidase（HRP） at 1.2 g/mL for varying times. MTT assay was applied to detect the cell proliferation. Flow cytometry was performed to detect the arrest of cell cycle. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） assay was used to measure apoptosis. 2, 7-dichlorofluorescin diacetate （DCFH-DA） uptake was measured to determine free radical by confocal microscope. Content of malondiadehyde （MDA） and activity of superoxide dismutase （SOD） were measured by biochemical methods. Results IAA/HRP initiated growth inhibition of K562 cells in a dose- and time-dependent manner. Flow cytometry revealed that cell cycle arrested at G1/G0 after 24 hours treatment. After 72 hours treatment, apoptotic rate of 100 mol/L IAA group increased to 43.9 %, which was 5 times that of control（P〈0.01）. Content of MDA and activity of SOD increased respectively in treatments compared with control. Meanwhile, IAA/HRP stimulated the formation of free radical, which was increased by IAA concentration-dependently. Conclusion The combination of IAAand HRP can inhibit the growth of Human leukemia cell line K562 in vitro by inducing apoptosis which is associated with the increase of free radical. The combination of IAA and HRP might be a promising chemopreventive and chemotherapeutic agent against human leukemia.     

IAA与HRP联合作用对人胰腺癌BXPC-3的细胞毒效应

目的　研究吲哚乙酸(IAA)与辣根过氧化物酶(HRP)在体外共同作用于人胰腺癌BXPC 3 细胞所产生的细胞毒效应。方法　MTT法测定IAA/HRP对BXPC 3细胞增殖的影响;流式细胞仪分析BXPC 3 细胞周期时相变化;胎盘兰(曲利本兰)排斥试验,研究IAA/HRP对BXPC 3 细胞死亡率的影响;原位细胞凋亡检测试剂盒检测IAA/HRP作用后BXPC 3细胞的凋亡。结果　IAA/HRP体外对BXPC 3 细胞有抑制生长的效应,且这种效应有浓度和时间依赖性;BXPC 3细胞在IAA/HRP作用48 h后细胞周期被阻滞在G1期和G2期;IAA/HRP作用72 h后对照组和药物组的凋亡细胞数存在着显著性差异,且凋亡细胞数有浓度依赖性。结论　IAA/HRP对胰腺癌BXPC 3 细胞有细胞毒作用,可体外抑制细胞的增殖,诱导凋亡。     

EXPLORE ON DIAGNOSTIC AND PROGNOSTIC VALUES OF EXTRAHEPATIC CHOLANGIOCARCINOMA： UTILITY OF SERUM CA19-9 AND SERUM CEA

Extrahepaticcholangiocarcinoma (EHCC)isamalignanttumorarisingfrombileductepithelium .Unlikemosthumancancers ,atissuediagnosisofcholangiocarcinomaisoftenextremelydifficultbe causeoftumorlocation ,size ,anddesmoplasticcharacteristics .Percutaneousfineneedleaspirationisfrequentlynotpossiblebecausemanyofthesetu morsarelocatedintheliverhilumamidlargevascu larstructure .Furthermore ,tumormassesareoftennotevenidentifiablebyCT ,ultrasound ,ormag neticresonanceimaging .Endoscopicapproachesarealsoofl…     

EFFECTS OF DESENSITIZATION AND REBOUND TO ADENOSINE ON ACTION POTENTIAL AND CONTRACTILITY IN ATRIAL CELLS IN GUINEA-PIGS

Ａｄｅｎｏｓｉｎｅ (Ａｄｏ)ｉｓａｎｅｎｄｏｇｅｎｏｕｓｐｕｒｉｎｅｎｕ ｃｌｅｏｓｉｄｅｔｈａｔｒｅｇｕｌａｔｅｓｍａｎｙｐｈｙｓｉｏｌｏｇｉｃａｌａｎｄｐａｔｈｏ ｐｈｙｓｉｏｌｏｇｉｃａｌｐｒｏｃｅｓｓｅｓｉｎｂｏｄｙ .Ｔｈｅｅｆｆｅｃｔｓｏｎｃａｒｄ ｉｏｖａｓｃｕｌａｒｓｙｓｔｅｍａｒｅｅｓｐｅｃｉａｌｌｙｏｂｖｉｏｕｓ .Ｐｅｏｐｌｅｆｏｕｎｄｔｈａｔａｃｅｔｙｌｃｈｏｌｉｎｅ (ＡＣｈ)ａｎｄａｄｅｎｏｓｉｎｅｈａｄｔｈｅｎｅｇａｔｉｖｅｉｎｏｔｒｏｐｉｃ ,ｎｅｇａｔｉｖｅｃｈｒｏｎｏｔｒ…     

顺铂对Hela细胞辐射增敏的体外实验观察

目的探讨顺铂对宫颈癌细胞系Hela的细胞毒性和辐射增敏作用,并研究射线与顺铂联合应用的效应关系。方法 MTT法检测Hela细胞在不同浓度药物和不同辐射剂量下的细胞抑制作用;辐射与顺铂联合应用时,MTT法检测细胞抑制率,绘制细胞抑制率曲线,研究顺铂与辐射联合应用时协同效应的条件;克隆形成法研究药物的辐射增敏作用,多靶单击拟合模型方程测定各组细胞辐射增敏比。结果顺铂作用于Hela细胞的半数致死质量浓度(IC50)为10μg/mL,药物与辐射共同作用于Hela细胞时,12 Gy辐射剂量下,顺铂1~5μg/mL对Hela的细胞毒性显示出明显的浓度依赖性和48 h内的时间依赖性,5μg/mL以上浓度顺铂对Hela的细胞毒性未见明显增加;集落形成法结果显示,顺铂辐射增敏比为1.320;辐射剂量小于10 Gy时,放射线与药物联合应用具有协同效应,而当辐射剂量大于10 Gy时,呈现相加效应或次相加效应。结论顺铂与辐射并用时对宫颈癌Hela细胞增殖具有显著抑制作用,药物具有最佳的辐射增敏剂量;且应尽早应用顺铂增敏化疗,以保证协同效应。     

ANTITUMOR MECHANISM OF GEM10 BY THE NATURAL KILLER ACTIVITY AND INTERLEUKIN-2 PRODUCTION

Studiesongermaniumcompoundsshowedthattheyhavemanybiologicaleffects ,suchasanti tu mor[1] ,improvingimmunity[2 ] ,reducingthecon centrationofbloodcholestrolandtrilycerides[3] ,in hibitingvirusreproduction ,butthesideeffectsoftoxicrestricttheirclinicaluses[4 ,5] .Therefore ,itisstillnecessarytodevelopothernewcompoundsoforganogermaniums.GeM 10 ,aneworganicgerma niumcompounds alkili metalcomplexcombinedwitheffectiveextractsofsomeChinesetraditionalherbs ,hasbeendemonstratedtobesaferandmorereliable.…     

THE INHIBITORY EFFECT OF MELATONIN ON THE GROWTH OF HUMAN BLADDER CARCINOMA T24 CELL LINE

Melatonin(Mel) ,anindoleamineprimarilyderivedfromthepinealgland ,issecretedduringthehoursofdarkness.Melactsasahormonalconduc tionof photoperiodinfluencedbytheseasonanddailyphysiologicalrhythms.Thefunctionofmelinhumansisobscure .However ,melhasattainedprominenceasatreatmentfordisturbedcircadianrhythmsandsleeppatterns ,hasenhancedthefunc tionofimmunitysystem[1] andhasinhibitedtumorgrowth[2 ] .Thepreviousstudieshaveindicatedthatmelcombinedwithotheranticancerdrugscanin creasetheefficiencyanddecre…     

FUCOIDIN INHIBITS OXIDIZED LOW DENSITY LIPOPROTEIN FROM INDUCING HUMAN PERIPHERAL BLOOD MONOCYTE EXPRESSION OF PROINFLAMMATORY CYTOKINES mRNA

Researcheshavealreadydemonstratedthatscav engerreceptorclassA(SR A)isabletobindoxidizedlowdensitylipoprotein(OxLDL)withhighaffinityandisoneofthemainreceptormediatingmurinemacrophagetouptakeOxLDL[1] .Itisalsoexpressedinhumanperipheralbloodmonocyte[2 ] andathero scleroticlesions[3 ] .However ,functionsofSR Ainmediatinghumanperipheralbloodmonocytetoup takeOxLDLaswellaspromotingtheatheroscleroticimmuno pathologicallesioninthelocalbloodvesselarestillunclear .Soforthepurposeofstudyingthefunc…     

增强型绿色荧光蛋白标记的Hela细胞株的建立

目的构建增强型绿色荧光蛋白(EGFP)标记的Hela细胞系。方法PCR和DNA测序鉴定pEGFP-C1真核表达质粒,采用Qiagen tip 500进行pEGFP-C1真核表达质粒DNA的提取和纯化;cellfectin转染Hela细胞,G418筛选,有限稀释法单克隆培养,荧光显微镜进行荧光检测EGFP的表达,采用激光共聚焦显微镜观测器检测荧光特性。结果PCR和DNA测序证实pEGFP质粒结构正确;在倒置荧光显微镜下,转染24h后,部分Hela细胞可见绿色荧光,转染72h时,大约80%Hela细胞内均可见绿色荧光。加入含G418的选择性培养基进行选择性培养,选择荧光较强的Hela细胞经有限稀释,持续筛选及克隆化培养,获得稳定表达绿色荧光的Hela细胞克隆,扩大培养并传至10代以上,将此细胞株命名为Hela-EGFP。激光共聚焦显微镜观察发现:在蓝光(-395nm)激发时,Hela-EGFP细胞绿色荧光激发波长为395nm,最大发射峰为509nm。结论Hela-EGFP细胞株具备稳定表达EGFP的能力,为实时可视化进行宫颈癌侵袭转移机制的研究奠定了基础。     

苦参素对Hela细胞增殖及凋亡的影响

目的探讨苦参素(Matrine)对体外培养的人宫颈癌Hela细胞增殖及凋亡的影响。方法Hela细胞体外培养,MTT法检测细胞增殖,TUNEL法检测细胞凋亡,免疫组化法检测细胞增殖核抗原(PCNA)蛋白的表达水平。结果与对照组相比,0.5、1.0、1.5、2.0、2.5、3.0g/L苦参素均能显著抑制HeLa细胞生长(P<0.05),药物浓度在0.5-1.5g/L之间时表现出时效和量效关系;苦参素2.0、2.5、3.0g/L与1.5g/L相比,抑制无统计学意义(P>0.05)。与对照组相比,1.0、1.5、2.0g/L苦参素组可见到显著性细胞凋亡(P<0.05);1.5g/L苦参素组作用48h和72h时PCNA蛋白表达显著下调(P<0.05)。结论苦参素可能通过抑制增殖和诱导凋亡的双重途径抑制宫颈癌Hela细胞的生长,苦参素有可能成为治疗子宫颈癌的一种药物。     

人宫颈癌细胞凋亡的研究

目的　观察顺铂、60 Co在宫颈癌细胞凋亡的作用及其引起的细胞形态学、DNA及细胞周期变化。方法　将人宫颈癌细胞株Hela,SiHa和RJC进行体外培养 ,用不同强度的放射线60 Co照射 ,观察照射后不同时间内肿瘤细胞形态变化 ,并采用末端脱氧核苷酰转移酶标记法 (TDT法 )检查细胞DNA有无断裂 ,用流式细胞仪记录细胞周期的改变和Bcl 2蛋白表达情况。同时将顺铂处理的宫颈癌细胞作为对照组 ,观察放射线和药物对宫颈癌细胞的联合作用。结果　经60 Co照射的三株细胞均出现凋亡的典型形态学改变 ,TDT法染色见受照细胞核内均有DNA断裂表现 ;细胞周期进展受到阻滞 ;Bcl 2蛋白表达在Hela细胞升高 ,而在SiHa和RJC细胞则降低 ,SiHa细胞对60 Co照射最为敏感 ,40 0CGy即可出现细胞凋亡的改变。顺铂单独处理可使宫颈癌细胞凋亡 ,与放射线联用有明显的增强细胞凋亡的效果。并在RJC细胞的G1 峰前出现凋亡峰。结论　宫颈癌细胞受到60 Co照射后 ,凋亡是其主要死亡形式 ,不同的宫颈癌细胞株对放射线的敏感性存在明显差异 ,与细胞凋亡有关的Bcl 2表达与宫颈癌细胞凋亡的发生呈负相关 ,放射线照射与化疗药物联合可以增加抑制宫颈癌生长的作用。     

EFFECTS OF TOTAL SAPONINS OF PANAX NOTOGINSENG AND LIGUSTRAZINE ON THE PROLIFERATION OF CEREBRAL MICROVASCULAR ENDOTHELIAL CELLS OF RATS

Ｅｎｄｏｔｈｅｌｉａｌｃｅｌｌｓ (ＥＣｓ)ｆｕｎｃｔｉｏｎａｓｔｈｅｂａｒｒｉｅｒｏｆｔｈｅｂｒａｉｎａｎｄｒｅｌａｔｅｔｏｔｈｅｓｅｃｒｅｔｉｏｎｏｆｓｏｍｅｅｎｄｏ ｃｒｉｎｅｓ.Ｔｈｅｉｒｄｙｓｆｕｎｃｔｉｏｎｉｓｃｌｏｓｅｌｙｒｅｌａｔｅｄｔｏｔｈｅｏｃｃｕｒｒｅｎｃｅａｎｄｄｅｖｅｌｏｐｍｅｎｔｏｆｃｅｒｅｂｒａｌｖａｓｃｕｌａｒｄｉｓｅａｓｅｓ[1] .ＤａｍａｇｅｔｏＥＣｓｉｓｔｈｅｆｉｒｓｔｓｉｇｎａｎｄａｂａ ｓｉｃｆａｃｔｏｒｏｆｃｅｒｅｂｒａｌｉｓｃｈｅｍｉａ .Ｔｈｅｒｅｆｏｒｅ ,…     

超声观测高同型半胱氨酸血症兔内皮功能损伤与血管细胞粘附分子的关系

目的研究高同型半胱氨酸血症(HHcy)兔血浆细胞粘附分子-1(VCAM-1)及一氧化氮(NO)的变化与内皮依赖性舒张功能(EDD)的关系,探讨内皮功能损伤的机理。方法将22只新西兰兔随机分为两组,每组11只,分别喂食正常饲料(正常对照组)、L-蛋氨酸[0.8g/(kg.d)]饲料(蛋氨酸组),分别于0、20、40、60、80d取血检测NO和VCAM-1的浓度,同时应用高分辨力超声检测其髂外动脉EDD的改变。结果①与正常对照组相比,蛋氨酸组兔髂外动脉的EDD显著降低,且呈明显的时间依赖性[80d,(-1.74±3.58)%vs.0d,(13.43±4.13)%,P<0.01];②与正常对照组相比,蛋氨酸组兔血浆中NO明显降低[80d,(217.43±27.38)μmol/L vs.0d,(379.75±56.83)μmol/L,P<0.05],血浆VCAM-1却明显升高[80d,(0.091 2±0.009 7)ng/mLvs.0d,(0.047 2±0.004 7)ng/mL,P<0.01],亦呈现明显的时间依赖性。结论高分辨力超声检测到的内皮依赖性舒张功能降低与血浆粘附分子明显相关,同型半胱氨酸(Hcy)导致内皮功能损伤是一个多途径的综合损伤。     

姜黄素联合顺铂对人卵巢癌SKOV-3细胞增殖的抑制作用

目的 探讨姜黄素及其联合顺铂对体外培养人卵巢癌SKOV-3细胞增殖的抑制作用.方法 将SKOV-3细胞培养并分别单独加入不同浓度姜黄素(5、10、20、40、80 μ mol/L)、顺铂(10 μmol/L)及联合作用后,用MTT法检测SKOV3细胞增殖情况.流式细胞术检测细胞凋亡率和细胞周期分布.结果 MTT法检测结果显示,不同浓度姜黄素组,随着药物浓度增加,作用时间延长,细胞的增殖抑制率上升(P＜0.05);联合用药组明显高于单一用药组(P＜0.05).流式细胞术结果显示,姜黄素能使卵巢癌SKOV-3细胞发生G2/M期阻滞,诱导细胞凋亡;随着姜黄素用药浓度增加,细胞凋亡率增高.结论 姜黄素对卵巢癌SKOV-3细胞增殖具有抑制作用,在一定范围内,这种抑制作用呈剂量-时间依赖性;姜黄素、顺铂联合应用具有协同作用.     

THE PROTECTIVE EFFECTS OF THE TOTAL SAPONIN OF DIPSACUS ASPEROIDES ON THE APOPTOSIS OF HIPPOCAMPAL NEURONS INDUCED BY β-AMYLOID PROTEIN

Objective To investigate the effects of the total saponin of Dipsacus asperoides (tSDA) and ginsenoside Rb1 (GRb1) on the apoptosis of primary cultured hippocampal neurons induced by β-amyloid protein (Aβ). Methods Primary cultured hippocampal neurons, the cultures were pretreated with tSDA and GRb1 on 10d for 24 hours respectively. Then the cultures were treated with 35μmol·L-1 Aβ25-35 for 24 hours, observed the changing of survival rate of neurons and the apoptosis of neurons with biochemical analysis combining immunofluorescent cytochemical double-staining technique. Results Hippocampal neurons were treated with 35μmol*L-1 Aβ for 24 hours, and survival rate of neurons downed to 52.6%. When neurons were pretreated by tSDA and GRb1, survival rate of neurons increased 11% to 15%. The findings of immunofluorescent cytochemical double-staining indicated that apoptotic neurons were obviously more than that of the blank group, reaching 43.9%.When neurons were pretreated by tSDA and GRb1, apoptotic neurons were downed to 16.6%, 10.8% respectively. Conclusion tSDA had the same effects as GRb1, protecting the neurons, antagonizing neurotoxicity of Aβ, increasing survival rate of neurons, and reducing apoptotic neurons induced by Aβ.     

中药安迪粉针剂对离体肿瘤细胞的细胞毒作用

目的　观察以蟾酥和人参为主要配方的中药粉针剂安迪 (Andi)对肿瘤细胞的药物毒性作用。方法　采用MTT法测定Andi对 3株人肿瘤细胞的细胞生长曲线、细胞存活率 (SR)和细胞倍增时间 (TD) ;观察Andi对ECA 10 9细胞生长的时效关系。克隆计数法检测不同浓度Andi对ECA 10 9细胞在含氧和乏氧状态下的生存分数 (SF) ,计算其乏氧细胞毒性比 (HCR)和半数抑制浓度 (IC50 )。结果　①Andi组和对照组对ECA 10 9、HepG2、SGC 790 1细胞的TD 分别为 118.72、12 3.0 8、4 0 .17h和 4 6 .12、4 0 .17、19.6 7h ;Andi组的SR分别为对照组的 33.33%、5 0 .5 4 %、36 .88%。② 12 .5 0、2 5 .0 0、5 0 .0 0 μg·mL- 1 Andi对ECA 10 9细胞在有氧和乏氧环境中的SF分别为 1.34、0 .97、0 .2 5和 0 .96、0 .2 7、0 .18。③Andi在乏氧环境中能抑制ECA 10 9细胞的生长 ,其作用大小与Andi浓度及作用时间呈正相关。④在有氧和乏氧下Andi的IC50 分别为 39.6 0 μg·mL- 1 和 2 3.5 0 μg·mL- 1 ,HCR为 2 .0 2。结论　Andi对多种人肿瘤细胞具有乏氧细胞毒作用 ,其程度与Andi的浓度和作用时间呈正相关     

PAC和PAFC在城市雨水处理中混凝效果研究

采用聚合氯化铝(PAC)、聚合氯化铝铁(PAFC)两种混凝剂对模拟城市雨水进行混凝试验研究,考察了混凝剂种类、投加量及原水pH等因素对混凝效果的影响.结果表明:混合搅拌强度200r/min快速搅拌1min,150r/min中速搅拌5min,50r/min慢速搅拌15min的条件下,PAC和PAFC的最佳投加量分别为40mg·L-1、60mg·L-1,两种混凝剂的混凝效果相当,无明显差别.考虑成本核算,选用PAC作为混凝药剂.在原水pH值为6.5~7.5的范围内PAC的混凝性能较好.在其投加量为40mg·L-1,原水pH为7.0的条件下,除浊率可达98%,除氨氮率为18%,CODcr的去除率达到47%.     

EFFECT OF ANGIOTENSIN Ⅱ RECEPTOR SUBTYPE ⅠON THE FIRING RATE IN CATECHOLAMINERGIC TUMOR CELLS

AngiotensinⅡ (AngⅡ ) playsanimportantroleintheregulationofbloodpressure[1 ] .Theac tionofAngⅡismediatedmainlybystimulationoftheangiotensintype 1(AT1 )receptorandinvolvesintheactivityofcatecholaminergicsystem .Im munostainingstudieshaveindicatedthattheAT1 re ceptorexistsonthecatecholaminergicneurons[2 ] .AdministrationofAngⅡintorat’sbrainincreasedtheconcentrationsofdopamine ,epinephrine ,andnorepinephrineinthebrain[3 ] .Althoughanatomicalandfunctionalevidencessupporttheinteractionbe twe…     

核桃壳对废水中Cr(Ⅵ)的静态吸附特性研究

采用废弃核桃壳对Cr(Ⅵ)浓度为20mg·L-1的模拟水样进行了静态吸附实验研究.实验结果表明,对于Cr(Ⅵ)浓度为20mg·L-1的50mL水样,当温度为25℃,采用粒径为1.0-1.6mm新疆核桃壳1.0 g、介质pH值为1.0、吸附时间为180min处理废水时,Cr(Ⅵ)的去除率可以达到99.3%.吸附后的水中C...