bcl—2反义核酸提高白血病HL60细胞对米托蒽醌的敏感性

目的 探讨下调HL60细胞bcl-2基因表达对米托蒽醌（MIT）细胞毒作用的影响。方法 将针对bcl-2基因编码区141-147密码子的反义寡核苷酸（ASODN）作用于HL60细胞,测定其mRNA表达,细胞生长。细胞凋亡。结果 12.5umol/LASODN作用于HL60细胞48h可以下调bcl-2mRNA表达40%。0.5umol/LMIT作用于经ASODN预处理48h的HL60细胞5h,50.9%的细胞发生凋亡,明显高于MIT单独作用诱导的细胞凋亡（25.6%);当ASODN与0.7nmol/LMIT共同作用时,细胞存活率（48h)由MIT单独作用时的73.7%降至25.6%。结论 bcl-2ASODN可下调bcl-2基因表达,促进细胞凋亡而增加MIT抗肿瘤作用。     

不同途径免疫的AFP基因修饰DC瘤苗体内抗肿瘤效应的研究

目的:评价bcl-2反义寡核苷酸对肾细胞癌细胞Bcl-2蛋白表达抑制及诱导凋亡作用.方法:合成与已知人类基因无同源性的bcl-2反义寡核苷酸(AS1与翻译起始端互补,AS2与编码区互补),以阳离子脂质体Lipofectin为转染载体,MTS比色实验测定细胞活率,反转录-聚合酶链反应(RT-PCR)测定bcl-2 mRNA的表达,Western杂交法测定Bcl-2蛋白表达,碘化丙啶(PI)染色流式细胞术测定凋亡细胞.结果:所测定的5个肾癌细胞株均有稳定的bcl-2 mRNA表达;转染的反义寡核苷酸对ACHN细胞Bcl-2蛋白的表达有明显抑制作用,而正义寡核苷酸无明显影响,AS2的抑制作用大于AS1;反义寡核苷酸可诱导ACHN细胞的凋亡,AS1和AS2的诱导率分别为32.1%和43.2%.结论:bcl-2反义寡核苷酸可抑制肾癌细胞Bcl-2蛋白的表达,并进而诱导细胞的调亡.     

全反式维甲酸诱导食管癌细胞凋亡的实验研究

目的探讨全反式维甲酸(ATRA)诱导食管癌EC9706细胞凋亡的作用及其机制。方法用不同浓度的ATRA处理EC9706细胞,采用四甲基偶氮唑盐(MTT)法检测ATRA对EC9706细胞的增殖抑制效应;采用流式细胞仪和原位末端标记(TUNEL)法检测细胞周期的变化和凋亡率;采用免疫组化S-P法,检测凋亡相关基因caspase-3和bcl-2的蛋白表达,并用病理图像分析软件进行半定量分析。结果ATRA对EC9706细胞有增殖抑制和诱导凋亡的作用;流式细胞仪检测显示,在G1期之前可出现Sub-G1峰,凋亡率最高为(32.6±0.4)%,并有浓度和时间依赖性;TUNEL法显示凋亡小体形成;EC9706细胞在凋亡过程中,凋亡相关基因caspase-3的蛋白表达增强,bcl-2的蛋白表达减弱。结论ATRA作用于食管癌EC9706细胞后将引起细胞凋亡的增加,这主要与ATRA能促进caspase-3的活化,并下调bcl-2的蛋白表达有关。     

天花粉蛋白对宫颈癌HeLa细胞凋亡及bax 、bcl-2 基因表达的影响

 目的 研究天花粉蛋白对人宫颈癌HeLa细胞的诱导凋亡作用并探讨其对凋亡相关基因bax和bcl-2表达的影响。方法 不同浓度天花粉蛋白分别处理HeLa细胞24～72h后，MTT法检测细胞的生长活性，显微镜观察细胞形态，流式细胞仪检测细胞周期，WesternBlot检测凋亡相关蛋白bax，bcl-2的表达。结果 显微镜下观察到细胞圆缩，染色体凝聚等凋亡细胞的形态学改变，流式细胞仪直方图上亦出现特征性的亚二倍体峰，随着天花粉蛋白浓度的增加，bax基因的表达增加而bcl-2的表达减少。结论 天花粉蛋白对HeLa细胞的生长具有抑制作用，可诱导宫颈癌HeLa细胞发生凋亡，bcl-2蛋白的减少与bax蛋白表达增多可能是HeLa细胞凋亡的机制之一。     

HoxB1质粒转染对视黄酸诱导的 HL-60细胞凋亡的影响

目的:探讨细胞内HoxB1表达、视黄酸(RA)代谢与细胞凋亡的相互关系.方法:采用脂质体介导的质粒转染和反转录-PCR等实验技术方法做有关分析检测.结果:2.5×10-7mol/L全反式RA(ATRA)处理HL-60细胞3天,HPLC分析显示,HoxB1的高表达可明显抑制HL-60细胞ATAR代谢;使ATRA增殖抑制作用减弱;野生型HL-60细胞有较强bcl-2表达,ATRA处理抑制未转染细胞bcl-2表达,但单纯HoxB1转染、HoxB1转染+IFNα均能增强bcl-2表达.结论:HoxB1转染HL-60细胞ATRA代谢明显受抑,ATRA增殖抑制作用及凋亡诱导作用减弱.     

bcl-2基因的反义寡核苷酸与三氧化二砷联合对恶性淋巴瘤细胞系凋亡的影响

目的：研究bcl-2基因的反义寡核苷酸与三氧化二砷（As2O3）联合对恶性淋巴瘤细胞系凋亡的影响。方法：采用细胞染色方法观察细胞凋亡的形态，原位末端标记法（TUNEL）检测凋亡细胞，流式细胞仪检测细胞的DNA含量和bcl-2蛋白的表达。结果：10－40μmol/l bcl-2基因的反义寡核苷酸和0．5－2．0μmol/L三氧化二砷均能抑制恶性B淋巴瘤细胞系Raji细胞生长，诱导细胞凋亡，流式细胞仪检测Raji细胞，发现亚G1期细胞明显增多，bcl-2蛋白的表达明显降低，呈现时间剂量依赖效应，两者联合应用比单独应用抑制作用显著。结论：bcl-2基因的反义寡核苷酸与三氧化二砷联合应用，明显抑制恶性B淋巴瘤细胞系生长，促进细胞凋亡。     

β-榄香烯诱导K562白血病细胞凋亡

目的 探讨β-榄香烯诱导人红白血病K562细胞株凋亡作用的机制。方法 采用Hoechst33342和PI荧光染色电镜、DNA电泳、免疫组化以及流式细胞术分析法,对K562细胞凋亡的定性与定量以及凋亡抑制基因编码蛋白bcl-2在肿瘤细胞内的表达进行检验。结果 经β-榄香烯处理的K562细胞出现核固缩,染色质边集, 凋亡小体形成,琼脂糖凝胶电泳呈现出细胞凋亡特征性的DNA梯状带,凋亡细胞发生率与药物浓度和作用时间呈正相关,即药物浓度为40ug/ml(48h),凋亡细胞的发生率从2.3%上升至58.4%,经β-榄香烯处理的K562细胞的bcl-2表达较对照组降低。结论 β-榄香烯对肿瘤细胞的杀伤主要是通过诱导细胞凋亡,其抑制细胞内Bcl-2的表达,是诱发肿瘤细胞凋亡的主要机制之一。     

环氧合酶-2抑制剂塞来昔布诱导人肝癌细胞凋亡及其机制

目的：探讨环氧合酶2（cyclooxygenase2,COX2）抑制剂塞来昔布（celecoxib）诱导人肝癌细胞系SMMC7721凋亡的作用及其可能机制。方法：以10、25、50、75、100 μmol/L的塞来昔布处理SMMC7721细胞,MTT法检测肿瘤细胞增殖,Hoechst 33342/PI 双染法检测细胞凋亡的形态特征,流式细胞术检测细胞凋亡率和细胞周期变化。RTPCR法检测Fas和Bcl2mRNA的表达,Western blotting检测细胞Fas和Bcl2 蛋白的表达。结果：塞来昔布以剂量依赖的方式抑制SMMC7721细胞增殖。塞来昔布作用后,SMMC7721细胞出现核染色质凝集、核膜破裂等凋亡细胞典型的形态特征。25、50和100 μmol /L塞来昔布诱导SMMC7721细胞的凋亡率分别为(16.32±2.32)%、(38.05±2.47)%、(71.17±3.19)%,并使细胞阻滞于G0/G1期。塞来昔布处理后,SMMC7721细胞Fas mRNA及蛋白表达明显增强（P<0.01）;Bcl2 mRNA表达无明显变化（P>005）,高浓度塞来昔布可下调Bcl2蛋白表达（P<0.01）。结论：塞来昔布能够抑制SMMC7721细胞增殖并诱导凋亡,其机制可能与影响凋亡相关基因表达有关。     

全反式维甲酸诱导人肝癌细胞HepG2凋亡的初步研究

[目的]研究全反式维甲酸（ATRA）体外诱导人肝癌细胞株HepG2的凋亡及其作用机制。[方法]ATRA处理HepG2细胞。MTT法分析细胞增殖反应;Hoechst染色法检测细胞凋亡情况;Western blot检测细胞中caspase-9、caspase-3和NF-κB蛋白表达情况。[结果]A-TRA可抑制HepG2细胞增殖并诱导细胞凋亡,可上调细胞中caspase-9、caspase-3表达水平（P〈0.05）。[结论]ATRA可抑制HepG2细胞增殖并诱导细胞凋亡,其作用机制可能与cas-pase-9、caspase-3表达上调有关。     

溶血磷脂酸抑制顺铂诱导的卵巢癌细胞凋亡

目的：探讨溶血磷脂酸（lysophosphatidic acid,LPA）对顺铂诱导的卵巢癌细胞凋亡的抑制作用及其作用机制。方法：体外培养卵巢癌细胞株SKOV3,采用MTT法检测LPA对顺铂（cisplatin,DDP）作用后卵巢癌细胞株SKOV3增殖活性的影响,Hoechst33258荧光染色观察凋亡细胞,用FCM法分析细胞周期变化和细胞凋亡率,DNA片段凝胶电泳观察凋亡细胞的DNA“梯状”条带,免疫细胞化学法及RT-PCR法分别检测细胞凋亡相关蛋白及其mRNA的表达。结果：LPA能降低DDP对SKOV3细胞生长的抑制作用,同时增加G0/G1期细胞比例,降低S期细胞比例和凋亡率。Hoechst33258染色检测示LPA作用后凋亡小体明显减少。DNA片段凝胶电泳示LPA作用后不产生明显的凋亡片段。10μmol/L的LPA作用SKOV3细胞48h后,bcl-2基因及其蛋白表达水平升高,而bax基因及其蛋白表达水平降低（P〈0.01）。结论：LPA可通过上调bcl-2基因及蛋白表达,下调bax基因及蛋白表达,抑制DDP诱导的卵巢癌细胞凋亡。因此,针对LPA的治疗有望提高DDP疗效,改善卵巢癌患者的预后。     

吡柔比星和阿霉素毒副反应的对比观察

 对含吡桑比星（THP）联合化疗方案施治的71例（183个治疗周期）和合阿霉素（ADM）联合化疗方案施治的106例（269个治疗周期）中晚期恶性肿瘤病人在毒副反应方面的资料进行了对比观察。结果表明，THP组的消化道副反应、脱发、心脏毒性较ADM组明显减轻；骨髓抑制、口腔炎、体温升高、肝肾功能损伤的毒副反应两药基本相当。综合判断，提示THP的毒副反应较ADM轻。     

In vitro evaluation of dichloro-bis(pyrazole)palladium(II) and dichloro-bis(pyrazole)platinum(II) complexes as anticancer agents

Introduction  Cisplatin (cis-diamminedichloroplatinum) was first identified for its anti-bacterial activity, and was later also shown to be an efficient anticancer agent. However, the therapeutic use of this anticancer drug is somewhat limited by its toxic side effects, which include nephrotoxicity, nausea, and vomiting. Furthermore the development of drug-resistant tumours is commonly observed following therapy with cisplatin. Hence there is a need for improved platinum derived drugs to overcome these limitations. Aims  Apoptosis contributes significantly to the cytotoxic effects of anticancer agents such as cisplatin; therefore in this study the potential anticancer properties of a series of pyrazole palladium(II) and platinum(II) complexes, [(3,5-R₂pz)₂PdCl₂] {R = H (1), R = Me (2)} and [(3,5-R₂pz)₂PtCl₂] {R = H (3), R = Me (4)}, were evaluated by assessment of their pro-apoptotic activity. Methods  The induction of apoptosis was measured in CHO cells by the detection of phosphatidylserine (PS) exposure using the annexin V and APOPercentage™ assays; DNA fragmentation using the Terminal deoxynucleotide transferase dUTP Nick End Labelling (TUNEL) assay; and the detection of activated caspase-3. Results  The platinum complexes were shown to be considerably more active than the palladium complexes, with complex 3 demonstrating the highest level of cytotoxic and pro-apoptotic activity. The LD₅₀ values for complex 3 and cisplatin were 20 and 70 μM, respectively, demonstrating that the cytotoxic activity for complex 3 was three times higher than for cisplatin. Various human cancer cell lines, including CaSki, HeLa, as well as the p53 mutant Jurkat T cell line were also shown to be susceptible to complex 3. Conclusions  Collectively, this in vitro study provides insights into action of palladium and platinum complexes and demonstrates the potential use of these compounds, and in particular complex 3, in the development of new anticancer agents.     

国产卡铂与顺铂治疗恶性卵巢肿瘤的疗效比较

采用国产卡铂和顺铂治疗恶性卵巢肿瘤６８例，Ⅰｃ期４例，Ⅱ期１１例，Ⅲ期３６例，Ⅳ期５例，复发转移病人１２例，随机分两组：卡铂用药组３７例，腹腔化疗２０例，全身化疗１７例；顺铂用药组３１例，腹腔化疗１８例，全身化疗１１例。卡铂治疗组有效率为５９％；顺铂对照组有效率为５２％，两组有效率相似（Ｐ＞０．０１）。卡铂腹腔化疗组有效率为６０％，略高于其他各组。卡铂组胃肠反应较顺铂组轻，血液系统变化与顺铂组无明显差别，对肾损伤较顺铂轻。卡铂可作为治疗晚期卵巢癌病人的一线药物。     

草酸铂联合氟尿嘧啶和亚叶酸钙治疗晚期大肠癌36例临床观察

目的 观察草酸铂(L-OHP)联合氟尿嘧啶(5-Fu)和亚叶酸钙(CF)治疗晚期大肠癌的近期疗效和毒副反应.方法 经病理组织学诊断的晚期大肠癌36例,用L-OHP 130 mg/m2,静脉滴注4h,每天1次;CF 200 mg,静脉滴注2 d,继以5-Fu 400 mg/m2,dl,d2,静脉滴入,5-Fu600 mg/m2,微量泵入A48 h以上,每3周重复1次,作为1个治疗周期,治疗2个周期后评定疗效.结果 36例总有效率为13例(37.1﹪),完全缓解1例(2.8﹪),部分缓解12例(33.3﹪),无变化14例(38.9﹪),进展9例(25﹪).主要的毒副作用为神经毒性、骨髓抑制、恶心呕吐等,无治疗相关性死亡患者.结论 L-OHP联合大剂量CF及5-Fu治疗晚期大肠癌疗效较好,毒副反应能为患者耐受,值得临床推广应用.     

Phase Ib dose-finding study of abiraterone acetate plus buparlisib (BKM120) or dactolisib (BEZ235) in patients with castration-resistant prostate cancer

BackgroundThe phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt) signalling axis and androgen receptor (AR) pathways exhibit reciprocal feedback regulation in phosphatase and tensin homologue (PTEN)-deficient metastatic castration-resistant prostate cancer (CRPC) in preclinical models. This phase Ib study evaluated the pan-PI3K inhibitor buparlisib (BKM120) and the dual pan-PI3K/ mammalian target of rapamycin (mTOR) inhibitor dactolisib (BEZ235) in combination with abiraterone acetate (AA) in patients with CRPC.Materials and methodsPatients with CRPC who had progressed on AA therapy received escalating doses of either buparlisib or dactolisib, along with fixed doses of AA (1000 mg once daily (qd)) and prednisone (5 mg twice daily (bid)). The primary objective was to define the maximum tolerated dose (MTD) and/or the recommended dose for expansion (RDE) of either buparlisib or dactolisib in combination with AA. Secondary objectives included safety, antitumour activity (Prostate Cancer Working Group 2 (PCWG2) criteria; 30% of prostate-specific antigen (PSA) decline at ≥week 12) and pharmacokinetic (PK) profile.ResultsIn buparlisib + AA arm, 25 patients received buparlisib + AA (median age, 67 years; Eastern Cooperative Oncology Group performance status (ECOG PS) of 0/1/2 for 7/17/1 patients, respectively). At 100 mg qd; two patients experienced dose-limiting toxicities (DLTs) (grade 3 hyperglycaemia; grade 2 asthenia), and this was the maximum buparlisib dose explored. Buparlisib + AA showed a 26% lower median area under the curve from time zero to 24°h (AUC_0–24) and 48% lower median maximum serum concentration (Cmax) versus the single-agent buparlisib assessed in first-in-human study. No objective response and few PSA decreases were reported.In dactolisib + AA arm, 18 patients (median age, 71 years; ECOG PS of 0/1 for 6/12 patients, respectively) received dactolisib + AA at the first dose level (200 mg bid). Five patients had 9 DLTs (grades 2&3 stomatitis; grade 3 hyperglycaemia; grades 2& 3 diarrhoea; grades 1& 2 pyrexia, grade 2 vomiting, and grade 2 chills).ConclusionsBased on the assessment of available pharmacokinetics, safety, and efficacy data, no further study is planned for either buparlisib or dactolisib in combination with AA in CRPC.     

癌胚抗原及铁蛋白联合测定在胃癌诊断中的价值

本文用放射免疫法测定58例胃癌、43例胃起性疾病、24例食管癌患者及158例健康成人血清与胃液中的CEA及Ft。结果表明,胃癌患者血清CEA水平高于正常对照组,P<0.01,但与胃良性疾病及食管癌纽问的差异不明显;胃癌患者血清Ft水平低于各对照组,P<0.01;胃癌患者的胃液CEA及Ft水平均高于各对照组,P<0.01。54例胃癌患者联合测定了血清CEA、胃液CEA及胃液Ft,胃癌胃液中的CEA、Ft双项阳性28例,敏感性为51.85％、特异性为99.04％、阳性诊断价值为96.55％。由此提示,胃液CEA及Ft联合测定有助于胃癌的诊断。     

子宫内膜癌组织中PCNA、MVD的检测及其意义

背景与目的:子宫内膜癌发病率呈逐年上升趋势,但其确切发病机制及预后因素尚不明确。本实验旨在研究增殖细胞核抗原(proliferating cell nuclear antigen.PCNA)指数、微血管密度microvessel density.MVD雪在子宫内膜癌组织中的表达,并结合肿瘤的手术病理分期、病理分级,进一步判断其价值。方法:采用免疫组织化学SP法,检测62例子宫内膜腺癌、22例正常子宫内膜组织的PCNA指数和MVD,结合肿瘤的临床分期、病理分级,对结果进行统计学分析。结果:子宫内膜癌组织中PCNA指数(54.49±9.29)和MVD(48.97±8.56)较正常子宫内膜明显增高(P<0.05);子宫内膜癌的期别越晚、分化越差,PCNA指数和MVD越高(P<0.05);在子宫内膜癌组织中,PCNA指数与MVD呈正相关。结论:PCNA指数、MVD增高与子宫内膜癌的发展有关;血管生成促进肿瘤细胞增殖。     

胃癌患者血浆TAFI及TAT的变化及其临床意义

目的 探讨胃癌患者血浆凝血酶激活的纤溶抑制物(thrombin activated fibrinolysis inhibitor,TAFI)和凝血酶-抗凝血酶复合物(thrombin-antithrombin complex,TAT)水平的变化及其与胃癌的关系.方法 收集原发性胃癌患者106例为实验组.同时,收集体检健康者30例为对照组.采用酶联免疫吸附双抗体夹心(ELISA)法定量测定血浆中凝血酶激活的纤溶抑制物抗原(TAFI:Ag)及TAT水平.结果 实验组血浆TAFI:Ag及TAT水平分别为(30.0±2.5)μg/mL及(36.3±3.4)ng/mL,对照组血浆TAFI:Ag及TAT水平分别为(22.5±2.5)μg/mL及24.5±2.5 ng/mL,有显著性差异(P<0.05).胃癌患者TAFI抗原及TAT水平随临床分期的进展而增高,Spearman秩相关分析提示,其与术后病理分期呈正相关,相关系数分别为0.814及0.881.用ROC曲线研究TAFI抗原和TAT两项指标的检验效能显示:TAFI抗原的临界值为26.25μg/mL(AUC=0.915,P<0.05);TAT的临界值为29.15 ng/mL(AUC=0.852,P<0.05).结论 胃癌患者体内TAFI及TAT水平均显著升高,并与胃癌病理分期呈正相关,提示其作为胃癌诊治的辅助实验室指标或有可观的应用价值.     

Distortion Product Otoacoustic Emission (DPOAE) as an Appropriate Tool in Assessment of Otoprotective Effects of Antioxidants in Noise-Induced Hearing Loss (NIHL)

Distortion product otoacoustic emission (DPOAE) appears to be an objective sensitive test of cochlear function. The aim of this study was to investigate whether DPOAE is an appropriate tool for assessment of minute changes in cochlea due to usage of antioxidant material. 48 workers exposed to continuous noise in a textile factory were randomly assigned into three groups: (1) The Control group (n = 16) received no antioxidant drugs, (2) The N-acetyl-cysteine (NAC) group (n = 16) received oral antioxidant NAC (1200 mg/day), (3) The Ginseng group (n = 16) received oral antioxidant Ginseng (200 mg/day). All three groups had a follow-up period of 2 weeks. The cochlear changes were assessed using DPOAE test before starting the daily work shift on first and 15th day. The associations between groups and DPOAE amplitudes after 2 weeks were analyzed using linear regression analysis. Four separate models were fitted by side of ears and frequency. All models were adjusted for baseline amplitude. Reduced (better) amplitude at DPOAE test was found for NAC and Ginseng groups at high frequencies (4 and 6 kHz) in both ears after 2 weeks compared to control group. Moreover, NAC group showed better DPOAE amplitude than Ginseng group. In conclusion, DPOAE seems to be an appropriate tool in assessing minute changes in the cochlea after antioxidant drugs administration.