赖氨大黄酸和紫杉醇对肺癌细胞增殖的抑制作用和凋亡诱导作用

目的:研究赖氨大黄酸（RHL）、紫杉醇单独和两者联合对人肺癌H460细胞增殖和凋亡的影响,阐明其作用机制。方法:选取处于对数生长期肺癌细胞株H460,随机分为对照组(不加药物)、紫杉醇组(1 μmol•L^-1紫杉醇)、RHL组(100 μmol•L^-1 RHL)和紫杉醇联合RHL组,并设空白组。采用MTT法和流式细胞术分别检测各组处理后48 h的细胞增殖率和凋亡率;采用Western blotting 方法检测凋亡相关蛋白和MEK/ERK蛋白的表达水平。结果:细胞培养48 h后,联合用药组细胞增殖率显著低于对照组、紫杉醇组和RHL组 (P<0.05),联合用药组细胞凋亡率显著高于对照组、紫杉醇组和RHL组(P<0.05),联合用药组caspase-3和poly ADP-ribose polymerase (PARP)的切割片段蛋白表达强度明显高于对照组、紫杉醇组和RHL组,联合用药组Bcl-2和NF-κB蛋白表达强度明显低于对照组、紫杉醇组和RHL组,同时RHL组紫杉醇上调的MEK和ERK蛋白磷酸化强度降低。结论:紫杉醇和RHL均可抑制肺癌H460细胞增殖,诱导细胞凋亡;RHL通过降低ERK活性、上调caspase-3和PARP的切割片段蛋白表达,增强紫杉醇抑制肺癌细胞增殖和凋亡诱导作用。     

重组人血管内皮抑制素联合化疗治疗晚期非小细胞肺癌近期疗效观察

目的观察重组人血管内皮抑制素注射液联合含铂化疗方案治疗晚期非小细胞肺癌(NSCLC)的客观疗效及安全性。方法18例Ⅲa～Ⅳ期NSCLC患者恩度TM(重组人血管内皮抑制素)注射液联合含铂化疗方案治疗2周期,其中恩度TM15mg静脉滴注14d,配合化疗一程为一周期,一周后重复。观察近期有效率、近期临床受益率、生活质量及毒副反应。结果近期有效率33.33%,近期临床受益率72.22%,生活质量有一定改善,毒副反应小。结论恩度注射液与含铂化疗方案联合应用治疗晚期非小细胞肺癌具有协同作用,可提高患者的近期疗效,改善患者的生活质量,未见明显的不良反应。     

原发性支气管肺癌与转移性肺癌血清T_3、T_4及TSH测定的临床观察

本文报告了20例原发性支气管肺癌患者与17例转移性肺癌患者血清T_3、T_4及TSH含量的变化。我们观察到原发性肺癌组血清T_8、T_4明显低于正常人组和转移性肺癌组(P<0.001),而TSH含量差异无显著性(P>0.05);转移性肺癌组与正常人组比较血清T_3降低(P<0.05),但T_4及TSH差异无显著性(P>0.05)。     

LY294002抑制小细胞肺癌干细胞样细胞自我更新

目的:研究Akt活性抑制剂LY294002抑制源自人小细胞肺癌NCI-H446细胞系肺癌球形成细胞即肺癌干细胞样细胞(LCSLCs)自我更新作用。方法:体外培养NCI-H446细胞系细胞。以干细胞条件培养基用超低粘附6孔细胞培养板悬浮培养富集和扩增LCSLCs。裸鼠皮下成瘤实验鉴别LCSLCs高致瘤特性。Western blot分析LCSLCs中Akt蛋白磷酸化水平。肿瘤球形成试验检测LY294002对LCSLCs自我更新的影响。结果:干细胞条件培养基悬浮培养6d,NCI-H446细胞系细胞呈三维非粘附性球体生长。LCSLCs具有高致瘤特性。与NCI-H446细胞系细胞比较,LCSLCs信号分子Akt组成性活化。LY294002有效降低LCSLCs中Akt磷酸化水平,并以剂量依赖方式抑制LCSLCs肺癌球形成(P<0.05)。结论:靶向干预小细胞肺癌LCSLCs信号分子Akt组成性活化可能成为抑制肺癌干细胞特性治疗小细胞肺癌的新策略。     

胸腔镜肺亚叶切除术治疗I期高龄非小细胞肺癌患者的回顾性研究

目的探讨胸腔镜肺亚叶切除术治疗I期高龄非小细胞肺癌患者的临床疗效。方法将行手术治疗且术后病理为I期非小细胞肺癌的126例年龄＞70岁的患者分为胸腔镜肺亚叶切除术组（59例）和胸腔镜肺叶切除术组（67例）,分析比较两组患者的年龄分布情况、术后并发症、术后引流时间、住院天数及术后随访情况。结果＞75岁患者多选择胸腔镜肺亚叶切除术。胸腔镜肺叶切除术患者总体并发症发生率（40.7%）显著高于胸腔镜肺亚叶切除术患者（19.4%）,差异有统计学意义（P=0.047）。胸腔镜肺亚叶切除术患者的平均术后引流时间、住院天数[（3.3±1.0）d、（8.5±1.5）d]短于胸腔镜肺叶切除术患者[（4.4±2.0）d、（12.8±2.0）d],差异均有统计学意义（均P＜0.01）。两组患者2、3年生存率的差异均无统计学意义（均P＞0.05）。结论胸腔镜肺亚叶切除术治疗老年I期非小细胞肺癌安全性良好,预后与肺叶切除术相似,可以作为老年I期非小细胞肺癌患者尤其是不能耐受肺叶切除者一个较好的选择。     

培美曲塞序贯吉非替尼对A549细胞凋亡的影响

目的 探讨培美曲塞序贯吉非替尼对A549细胞的疗效,及其对A549细胞增殖、凋亡及凋亡相关蛋白表达的影响.方法 采用MTT[3-（4,5-二甲基噻唑-2）-2,5-二苯基四氮唑溴盐]比色法检测培美曲塞、吉非替尼、培美曲塞联合吉非替尼及培美曲塞序贯吉非替尼对A549细胞增殖的影响;Western blot 检测凋亡相关蛋白的表达.DAPI核染色法检测A549细胞凋亡的形态学变化.结果 培美曲塞序贯吉非替尼对A549细胞增殖的抑制作用较单药及两药联合治疗更明显,A549细胞中培美曲塞、吉非替尼作用72 h的半数抑制浓度IC50分别为0.84 μmol/L和3.35 μmol/L.Western blot检测到培美曲塞序贯吉非替尼组与培美曲塞、吉非替尼单药治疗组比较,Bcl-2、Caspase3、Caspase8及PARP蛋白表达明显降低,Bax、p-H2AX蛋白表达明显升高,差异有统计学意义（P〈0.05）.DAPI核染色后可观察到培美曲塞序贯吉非替尼诱导A549细胞72 h后凋亡明显增强.结论 培美曲塞、吉非替尼均可明显抑制A549细胞增殖,诱导A549细胞凋亡,培美曲塞序贯吉非替尼作用更明显.     

联合应用缺血后处理、远隔缺血后处理和纳洛酮后处理对大鼠脑缺血-再灌注损伤的影响

Objective To assess the effects of ischemic postconditioning, remote ischemic postconditioning and naloxone postconditioning on focal cerebral ischemia-reperfusion injury in rats.Methods A total of 110 adult SD rats were randomly divided into 5 groups (n =22 each). The focal cerebral ischemia-reperfusion injury was induced by a 90-minute occlusion of right middle cerebral artery (MCA) and a 24-hour reperfusion sequentially. Group 1 was of ischemia-reperfusion control; Group 2 ischemic postconditioning induced by three 30-second cycles of MCA occlusion followed by a 30-second reperfusion; Group 3 remote ischemic postconditioning performed via a transient occlusion of right femoralartery at 5 min before the initiatlon of reperfusion:Group 4 naloxone posteonditioning with naloxone 10 mg/kg intraperitoneaUy injected at the initiation of reperfusion;Group 5 combined ischemic,remote ischernic & naloxone postconditioning performed simultaneously in accordance with the methods used in Groups 2,3 & 4.The neumlogie deftcit scores(NDS)were obtained at 2 h & 24 h post-reperfusion.At 24 h post-reperfusion.the anesthetized rat was sacrificed by decapitation and the brain rapidly extracted to asseSS the size ofcerebral infaret(n=10),detect the cerebral expression of microtubule-associated protein2(MAP2)(n=6),measure the plasma volume of cerebral tissues and quantify the diameter and segment artery at 5 min before the initiation of reperfusion; Group 4 naloxone postconditioning with naloxone 10 mg/kg intraperitoneally injected at the initiation of reperfusion; Group 5 combined ischemic, remote ischemic & naloxone postconditioning performed simultaneously in accordance with the methods used in Groups 2, 3 & 4. The neurologic deficit scores ( NDS) were obtained at 2 h & 24 h post-reperfusion. At 24 h post-reperfusion, the anesthetized rat was sacrificed by decapitation and the brain rapidly extracted to assess the size of cerebral infarct (n = 10), detect the cerebral expression of microtubule-associated protein2 ( MAP2) (n =6) , measure the plasma volume of cerebral tissues and quantify the diameter and segment length of cerebral microvessel (n = 6 ). Results There were no significant differences in the heart rate (HR) and mean arterial pressure (MAP) among the above five groups at all observed time points (P ＞ 0. 05). At 24 h post-reperfusion, the percentage of ischemic cerebral infarct size was 43% ±6% , 31% ±4% , 32% ±5% , 28% ±6% & 21% ±7% in ipsilateral hemisphere area (i. e. , cerebral infarct severity)in Groups 1-5 respectively. Compared with Group 1, the levels of NDS and cerebral infarct severity significantly decreased at ischemic side in Groups 2-5 ( P ＜ 0. 05 ). And the cerebral expression of MAP2,plasma volume of cerebral tissues, diameter and segment length of cerebral microvessel significantly increased at the ischemic side (all P＜0. 05). However, there were no significant differences in the abovementioned parameters at ischemic side among Groups 2, 3 and 4 (all P ＞0. 05). The parameters of NDS,cerebral infarct severity, cerebral expression of MAP2 and plasma volume of cerebral tissues in the ischemic side significantly increased in Group 5 compared with Groups 1,2,3 and 4 (all P ＜ 0. 05). The diameter and segment length of cerebral microvessel at ischemic side were not different among Groups 2,3,4 and 5 (all P＞0. 05). Conclusion In focal cerebral ischemia-reperfusion rats, ischemic, remote ischemic and naloxone postconditioning may produce significant neuroprotective effects of reduced cerebral infarct severity and improved neurologic dysfunctions. A combination of three postconditioning approaches enhances the above neuroprotective effects.     

清热解毒药配伍桔梗汤对急性肺损伤模型大鼠TLR4 mRNA表达的影响

目的 观察清热解毒中药配伍桔梗汤对内毒素致急性肺损伤(ALI)模型大鼠肺Toll样受体4(TLR4) mRNA表达的影响,探讨桔梗汤的配伍增效机制.方法 雄性SD大鼠按随机数字表法分为生理盐水组、ALI模型组、清热解毒药组、清热解毒药加桔梗汤组.股静脉注射内毒素脂多糖制备ALI大鼠模型,检测各组大鼠血清肿瘤坏死因子-α(TNF-α)、白细胞介素-10(IL-10)含量及肺组织髓过氧化物酶(MPO)活性、TLR4 mRNA的表达.结果 与生理盐水组相比,ALI模型大鼠血清TNF-α、IL-10含量明显升高(P<0.01或P<0.05),肺组织MPO活性、TLR4 mRNA表达明显增强(P<0.01或P<0.05).中药干预后大鼠血清TNF-α、IL-10含量及肺组织MPO活性、TLR4 mRNA表达均明显下降(P<0.01或P<0.05),且清热解毒药与桔梗汤配伍疗效优于单纯清热解毒药(P<0.05).结论 桔梗汤对清热解毒药降低血清TNF-α、IL-10含量、抑制MPO活性及TLR4 mRNA基因表达、发挥解毒抗炎功效,具有促进作用.     

不同缺血预处理方式对幼兔心肌低温缺血/再灌注损伤的影响

目的探讨缺血预处理(ischemicpreconditioning,IP)对兔未成熟心脏低温缺血/再灌注损伤的影响及其可能机制。方法采用Langendorff离体心脏灌注模型,3~4周龄幼兔心脏30只,随机分为缺血预处理1组(IP1组)、缺血预处理2组(IP2组)、格列苯脲1组(G1组)、格列苯脲2组(G2组)及对照组(Con组)5组,每组6只。5组幼兔心脏均以95%O2 5%CO2混合气体饱和的K-H液常温平衡灌注30min后:Con组续灌20min;IP1、IP2组心脏分别给予1次、2次IP,每次IP由5min缺血和5min再灌注组成;G1、G2组给予格列苯脲10μmol/L后分别给予1次、2次IP。然后各组心脏均在20℃低温下缺血120min,37℃常温下再灌注30min。以MacLabV/4S生理实验系统记录平衡末、缺血前及再灌注后1、3、5、10、20、30min时左心功能指标,包括左心室发展压(LVDP)、左心室内压上升最大速率( dp/dtmax)、左心室内压下降最大速率(-dp/dtmax)和心率(HR);测定再灌注末心肌组织中ATP和丙二醛(MDA)含量,超氧化物歧化酶(SOD)及Ca2 -ATP酶的活性。结果再灌注10min后,IP2组各对应时间点的LVDP×HR、±dp/dtmax的恢复百分比均显著高于Con组(P<0.01,P<0.05);再灌注末IP1、IP2组心肌组织的ATP含量及Ca2 -ATP酶的活性均显著高于Con组(P<0.01,P<0.05),IP1组的MDA含量明显低于Con组(P<0.01);G1、G2组各指标与Con组相比无显著差异(P>0.05)。结论IP对低温缺血的兔未成熟心脏具有一定的心肌保护作用,其效应与IP的方式有关,其机制可能与KATP有关。