He-Ne激光血管内照射对脑梗死患者血液流变学和细胞凋亡的影响

目的探讨He-Ne激光血管内照射(ILLLI)对脑梗死患者疗效、血液流变学性质、淋巴细胞增殖、凋亡和染色体的影响。方法脑梗死患者70例随机分成对照组和治疗组,各35例。对照组采用常规药物治疗,治疗组在常规药物治疗的基础上加用He-Ne激光血管内照射治疗,比较两组疗效和治疗前后的血液流变学指标、细胞增殖指数(MI)、细胞周期动力学指数(CKI)、姐妹染色单体交换(SCE)频率及淋巴细胞凋亡频率。结果治疗组有效率88.6%,对照组有效率65.7%,差异有非常显著意义(P<0.05)。治疗组全血黏度(低切,高切)、毛细血管血浆黏度,血沉、血沉方程K值、红细胞聚集指数、红细胞刚性指数较治疗前明显下降,红细胞变形指数较治疗前增加,治疗前后差异有显著意义(P<0.05)。治疗组患者ILLLI后淋巴细胞增殖活性高于对照组(P<0.05),CKI与对照组相比差异无显著意义(P>0.05);两组间SCE频率差异无显著意义(P>0.05);治疗组脑梗死患者的淋巴细胞凋亡率明显高于对照组(P<0.001),也明显高于治疗前(P<0.001)。结论低强度He-Ne激光血管内照射能有效改善血液流变学性质,改善微循环,增加脑血流量,促进神经功能的恢复,显著提高脑梗死患者的疗效,并能提高脑梗死患者的淋巴细胞增殖活性;可诱导发生较高的淋巴细胞凋亡率,对细胞没有致突变性。     

Toll样受体4对肿瘤细胞放射敏感性的影响

目的 观察Toll样受体4(TLR4)对肿瘤细胞放射敏感性的影响.方法 将体外培养的RAW264.7、Lewis、MFC、Hepal-6、B16和NIH3T3细胞各分为假照组和5 Gy照射组,采用流式细胞术检测照射24 h后TLR4表达变化,从中筛选出照射后高表达和低表达TLR4的细胞,将其各自分为TAK242阻滞组、LPS刺激组和空白对照组,采用CCK-8试剂盒检测其增殖活性,用克隆形成实验计算其细胞存活分数,用Annexin V凋亡试剂盒检测其凋亡率,用流式细胞术观察细胞周期进程的变化.结果 5 Gy照射24 h后Lewis细胞TLR4表达水平明显高于照射前(t=-8.68,P＜0.01),MFC细胞则相反,照射后TLR4表达水平明显低于照射前(t=25.80,P＜0.01),而RAW264.7、Hepa1-6、B16细胞的TLR4表达水平比照射前有升高趋势,但无明显差异.5 Gy照射后Lewis和MFC细胞的增殖活性均明显降低(t=57.62、-6.23,P＜0.01),而用TAK242阻滞TLR4后Leiws细胞增殖活性更加降低(t=5.96,P＜0.01),但MFC细胞增殖活性反而明显升高(t=4.16,P＜0.01).5 Gy照射后Lewis和MFC细胞的存活分数均明显降低(t=13.37、19.24,P＜0.01),用TAK242阻滞TLR4后Leiws和MFC细胞存活分数更加降低(t=4.90、4.42,P＜0.05).5 Gy照射后Lewis细胞的凋亡率明显高于假照组(t=-167.85,P＜0.01),阻断TLR4后明显高于单纯照射组(t=-4.73,P＜0.01).照射后MFC细胞的凋亡率升高(t=-26.45,P＜0.01),阻断和激动TLR4其凋亡率均显著低于单纯照射组(t=8.87、-3.05,P＜0.05).Lewis和MFC细胞受照后G0/G1期、S期细胞明显降低(t=8.68、14.80、20.31、4.48,P＜0.01),G2/M期细胞上升(t=-37.48、-13.06,P＜0.01).两种细胞的TAK242阻断组和LPS刺激组各周期进程均无明显变化.结论 Lewis细胞的TLR4高表达与其受照后的增殖活性和凋亡率变化有关,而与其周期进程无关,TLR4影响肿瘤细胞的放射敏感性.     

乙醇降低人乳腺癌MCF-7细胞放射敏感性的研究

目的 研究乙醇对人乳腺癌MCF-7细胞放射敏感性的影响及相关机制.方法 将人乳腺癌MCF-7细胞分为4组,对照组(不做任何处理)、乙醇组(乙醇处理)、单纯照射组(6 GyX射线处理)和联合组(乙醇与X射线联合作用).克隆形成法检测50或100 mmol/L乙醇对MCF-7细胞放射敏感性的影响;流式细胞术检测细胞周期变化;Annexin V-FITC法检测细胞凋亡.结果 50和100 mmol/L乙醇处理MCF-7细胞50 h,对生长无明显影响(t--0.82和1.15,P＞0.05);乙醇预处理2h,可明显增加X射线照射后MCF-7细胞克隆形成的能力(t=4.15和10.28,P＜0.05).相比于单纯照射组,乙醇降低了X射线照射诱导的细胞G2/M期阻滞(t=7.18,P＜0.05),以及sub-G1 峰的比例(t =5.39,P＜0.05).而且,Annexin V-FITC检测结果示,乙醇联合X射线照射的细胞晚期和早期凋亡减少(t=4.86和7.59,P＜0.05).结论 乙醇可以增加MCF-7细胞的辐射抗性,其机制可能与降低辐射诱导的细胞G2/M期阻滞及早、晚期凋亡的发生相关.     

Ad-Rb94联合照射对体外人大肠癌细胞的抑瘤作用

目的 探讨人视网膜母细胞瘤Rb94基因重组腺病毒载体(Ad-Rb94)联合γ射线照射对体外人大肠癌细胞生长的联合抑瘤作用及其机制.方法 将Ad-Rb94于体外转染大肠癌HT29细胞,转染后12h进行4Gy 137Csγ射线照射.实验分组为5组:对照组、β-半乳糖苷酶基因重组腺病毒载体组(Ad-lacZ组)、Ad-Rb94组、照射组和Ad-Rb94联合照射组.用噻唑蓝法检测HT29细胞的生长,用流式细胞术检测HT29细胞的细胞周期和细胞凋亡的变化.结果 当Ad-Rb94有效转染HT29细胞后,从转染第4日开始,照射组和联合照射组细胞生长速率较对照组和Ad-lacZ组缓慢;与Ad-Rb94组和照射组相比,Ad-Rb94联合照射组细胞的生长表现出更强的抑制效应(t=15.02、17.30,P＜0.01).细胞周期结果表明,与对照组、Ad-lacZ组和Ad-Rb94组相比,照射组大量细胞停留在G2期,各组间差异有统计学意义(t=18.65、15.23、16.38,P＜0.01);但Ad-Rb94联合照射组停留在G2期细胞更多(约40％),远高于照射组(t=7.78,P＜0.05).细胞凋亡结果表明,与对照组相比,Ad-Rb94组和照射组细胞凋亡率明显增加(t=16.19、10.72,P＜0.01);Ad-Rb94联合照射组细胞凋亡率最高(21％),与Ad-Rb94组和照射组相比,差异有统计学意义(t=6.17、9.25,P＜0.05).结论 腺病毒介导的Rb94基因联合照射对大肠癌细胞的抑瘤作用具有协同效应,其机制可能是促进细胞G2期阻滞和凋亡.     

表没食子儿茶素没食子酸酯对人鼻咽癌CNE-1细胞辐射敏感性的影响

目的 探讨表没食子儿茶素没食子酸酯(EGCG)对人鼻咽癌CNE-1细胞辐射敏感性的影响及相关机制.方法 将细胞分为对照组、EGCG单独处理组、UVC或X射线单独照射组,EGCG联合UVC或X射线照射组.四甲基偶氮唑盐(MTT)比色法检测不同浓度EGCG处理24、48和72 h对CNE-1细胞生长的影响,以及EGCG联合紫外线(UVC)及X射线对CNE-1细胞生长的影响.克隆形成实验检测EGCG对CNE-1细胞的放射增敏作用.流式细胞术检测细胞周期变化.Annexin V-FITC法检测细胞凋亡.Western blot法检测相关蛋白表达水平.结果 EGCG可抑制CNE-1细胞的生长,并有剂量·效应关系(r=0.817)和处理时间-效应关系(r=0.364).EGCG在低剂量50μmol/L,预处理2h,可增加CNE-1细胞对UVC和X射线的辐射敏感性.相比于单独照射组,EGCG联合UVC照射组细胞的S期阻滞消失(t=18.68,P＜0.05),sub-G1峰的比例则明显增高(t =6.67,P ＜0.05).而且,Annexin V-FITC检测结果揭示,EGCG联合UVC照射组的细胞中早期凋亡细胞比例显著增加(t=10.28,P ＜0.05).但与单独照射组细胞相比,EGCG联合X射线照射组细胞的S期及G2/M期阻滞更明显(t=7.11和6.99,P＜O.05),无明显的sub-G1峰出现.EGCG联合UVC照射可使凋亡相关蛋白Bax表达增加(t=5.42,P ＜0.05),Caspase-3表达增加(t=4.14,P＜0.05),Bcl-2的表达变化并不明显(t=1.85,P＞0.05),而其联合X射线对于细胞周期相关蛋白Cdc2 p34表达的影响不大(t=1.04,P＞0.05).结论 EGCG可抑制鼻咽癌CNE-1细胞的生长.EGCG与UVC联合损伤作用与促进细胞发生早期凋亡相关,涉及到凋亡相关蛋白Bax和Caspase-3;而EGCG和X射线的联合损伤作用可能与周期阻滞相关.     

C/EBP同源蛋白在严重创伤性脑损伤患者中的表达

目的 研究C/EBP同源蛋白(C/EBP homology protein,CHOP)在严重创伤性脑损伤(traumatic brain injury,TBI)患者颅脑周围组织中的表达,探讨其与损伤严重程度的关系. 方法 选取41例TBI患者(TBI组)颅脑周围组织,并选取因其他疾病(排除TBI及其他中枢神经系统疾病)死亡的16例尸检标本作为对照组,对照组及TBI组按年龄分为未成年组(≤18岁)、成年组(18 ～59岁)和老年组(＞59岁).TBI组患者根据入院时GCS评分再分为重型TBI组(GCS 6 ～8分)和特重型TBI组(GCS 3～5分).比较不同年龄、性别及GCS组间CHOP在TBI周围组织中的表达差异.TUNEL法检测神经细胞凋亡,并对CHOP表达水平与凋亡数量进行相关性分析. 结果 对照组CHOP表达在年龄与性别间差异无统计学意义(P＞0.05).与对照组比较,TBI组CHOP表达显著上调(P＜0.05).在TBI组中,CHOP表达在性别间差异无统计学意义(P＞0.05),而老年组患者CHOP表达低于成年组和未成年组(P＜0.05),特重型TBI组明显高于重型TBI组(P＜0.05).TBI组较对照组神经细胞凋亡数量明显增加(P＜0.05),且CHOP表达水平与凋亡指数呈正相关(r =0.72,P＜0.05). 结论 TBI后CHOP表达水平与损伤严重程度及神经细胞凋亡密切相关,但CHOP诱导的凋亡途径可能不是老年组TBI后继发性脑损害的主要因素.     

反义端粒酶反转录酶对5-羟色胺诱导的肺动脉平滑肌细胞增殖抑制的机制研究

目的 探讨端粒酶反转录酶(TERT)的反义寡核苷酸(ASODN)在抑制5-羟色胺(5-HT)诱导的肺动脉平滑肌细胞(PASMC)增殖过程中对细胞凋亡和细胞周期的影响.方法 组织块法培养大鼠PASMC,并将细胞分为对照组(应用RPMI-1640培养液培养)、5-HT组(应用含10-5mol/L 5-HT的培养液培养)、反义核酸组(应用含5-HT、反义TERT寡核苷酸的培养液培养)和正义核酸组(应用含5-HT、正义TERT寡核苷酸的培养液培养).采用Hoechst染色、免疫荧光显微镜检测各组细胞凋亡形态,并采用流式细胞仪检测细胞凋亡和细胞周期,免疫组化法检测细胞核增殖抗原(PCNA)在细胞中的表达.结果 Hoechst染色实验显示5-HT组凋亡细胞数(161±33)明显高于对照组(63±16,P＜0.05),而与正义核酸组(177±48)比较差异无统计学意义(P>0.05);反义核酸组凋亡细胞数(289±58)高于5-HT组和正义核酸组(P＜0.05).流式细胞仪检测提示5-HT可促进正常细胞的早期和晚期凋亡,并促使细胞由G1期向S期转化.反义TERT对5-HT引起的细胞凋亡有协同作用,并引起G期阻滞.免疫组化实验显示,5-HT组PASMC的PCNA表达较对照组增加(P＜0.05);正义核酸组PCNA表达高于对照组(P＜0.05),并与,5-HT组表达相近;而反义TERT组PCNA的表达显著低于5-HT组和正义核酸组(P＜0.05).结论 反义TERT可有效抑制5-HT诱导的PASMC增殖,为肺动脉高压的基因治疗提供了理论基础.     

超声诊断剂量与人早孕绒毛细胞凋亡

目的检测超声诊断剂量与人早孕绒毛细胞凋亡的关系.材料与方法将拟进行人工流产的24例早孕(45～60d)妇女随机分为4组对照组为空白照射组、10min组、20min组和30min组.应用Hp 8500彩色多普勒超声诊断仪,对孕囊进行不同时间的持续照射,照射后24h取材.用原位末断脱氧核苷酸转移酶标记(TUNEL)技术检测上述各组绒毛组织凋亡细胞.结果超声照射后10min组绒毛组织凋亡细胞与对照组无差别,20min组和30min组凋亡细胞明显增加,显著大于对照组和10min组(P＜0.001).结论诊断超声持续照射早孕孕囊组织＞10min可引起绒毛滋养层细胞凋亡增加,且随照射时间延长而继续增加.     

低强度激光照射对大鼠血管平滑肌细胞一氧化氮生成通路的上调作用

目的 观察低强度激光照射(low power laser irradiation,LPLI)对大鼠血管平滑肌细胞(vascular smooth muscle cells,VSMCs)左旋精氨酸(L-Arg)转运、一氧化氮(NO)生成以及NO合酶(nitric oxide synthase,NOS)活性的作用,以探讨LPLI血管保护作用的细胞分子机制.方法 取Wistar大鼠主动脉,贴块法培养VSMCs,分别以能量密度0、0.9.2.7和4.5 J/cm2的He-Ne激光(波长632nm)照射,无照射组为对照组.重氮化学反应法测定培养液中NO含量,细胞孵育液中加入3H-L-Arg测定细胞L-Arg的摄入,测定3H-胍氨酸的生成反映NOS活性.结果 能量密度分别为0.9,2.7和4.5 J/cm2的LPLI照射VSMCs,促进细胞NO生成分别较对照组增加-1.3%(P＞0.05)、60.4%(P＜0.01)和78.6%(P＜0.01);细胞NOS活性分别较对照组增加16.9%(P＞0.05)、44.6%(P＜0.01)和53.0%(P＜0.01);而且细胞摄入L-Arg(Vmax值)分别较对照组增加11.0%(P＜0.05),66.7%(P＜0.01)和38.7%(P＜0.01);激光照射对L-Arg跨细胞膜转运的米氏常数(Km值)无明显影响(均P＞0.05).结论 低强度He-Ne激光照射激活大鼠VSMCs的L-Arg/NOS/NO通路,可能为低强度激光扩张血管、改善器官血流等生物学效应的机制之一.     

系统性红斑狼疮患者外周血淋巴细胞Bc1-2蛋白表达

目的红斑狼疮与细胞凋亡障碍有关,Bc1-2通过干扰细胞凋亡从而延长细胞寿命,因此,探讨Bc1-2在SLE发病中的作用十分重要.方法运用免疫组化法测定Bc1-2在63例SLE患者外周血淋巴细胞的表达.结果SLE患者外周血淋巴细胞Bc1-2的表达明显高于正常对照组(P＜0.01),活动期与缓解期相比亦有较显著性意义(P＜0.05).结论外周血淋巴细胞凋亡可能是SLE的发病机理之一.     

严重急性呼吸综合征肺脏及肺外器官细胞凋亡及其机制的研究

目的 探讨SARS冠状病毒(SARS-CoV)诱导的细胞凋亡在SARS发病机制中的意义。方法 采用原位末端标记(TUNEL)法与Cytokeratin(CK)及CD3、CD8、CD20、CD68单克隆抗体免疫组化双染色方法,对6例SARS死亡患者的肺及肺外组织进行多种细胞凋亡状况的研究,并以免疫组化法检测凋亡相关蛋白Fas、Fas配体(FasL)、Bcl-2和P53的表达。结果 与正常组织比较,SARS患者全身多器官组织细胞凋亡数目明显增多,尤以肺脏和免疫器官为甚,主要的凋亡细胞类型为CK阳性肺泡上皮细胞、终末支气管黏膜上皮细胞、CD3细胞、CD8细胞,以及少部分CD20’细胞和CD68’巨噬细胞。Fas蛋白主要于SARS肺组织内浸润的单个核细胞胞膜及胞质内表达,而FasL主要分布于SARS-CoV感染的靶细胞,尤其是上述凋亡细胞内,肺及免疫器官内少数细胞亦表达FasL,但Bcl-2及P53表达明显下调。结论 SARS-CoV诱导的广泛且迅速的细胞凋亡可能为SARS患者肺脏及免疫器官的主要细胞损伤机制之一;SARS-CoV诱导的主要凋亡细胞与体内SARS-CoV主要的感染靶细胞相吻合;表达Fas的致敏T淋巴细胞与表达FasL,的靶细胞结合介导的细胞凋亡可能为SARS-CoV诱导细胞凋亡的重要机制之一,而Bcl-2和P53的表达下调可能参了细胞凋亡过程。     

12C重离子束对人淋巴细胞增殖、周期和凋亡的影响

目的 探讨¹²C重离子束对人淋巴细胞增殖以及周期、凋亡的影响.方法 ¹²C重离子束照射人淋巴细胞Peng-EBV,吸收剂量分别为0(对照组)、0.5、2.0 Gy.照射后用MTS法检测细胞增殖活力,流式细胞仪检测细胞周期和细胞凋亡.结果 与对照组相比,0.5 Gy照射可以增加细胞增殖活力(t=2.66~14.45, P<0.05),而2.0 Gy照射降低了细胞活力(t=7.65~64.45, P<0.05).受照射细胞的活力存在一个恢复和下降过程,受照后48 h内,细胞数量呈增加趋势,但72 h时细胞数量下降.照射后48 h,两组细胞G₂/M期呈明显上升趋势,高于对照组(t=2.01~99.80,P<0.05),且2.0 Gy组的周期阻滞较0.5 Gy组严重;照射后30 d,细胞周期阻滞恢复到正常水平.照射后12、24、48 h,两照射组与对照组相比,细胞凋亡率差异有统计学意义(t=-3.05~-1.05,P<0.05),在照后24 h最高,48 h明显下降,30 d恢复到对照水平.结论 ¹²C离子束辐射影响人淋巴细胞增殖,诱导人淋巴细胞发生明显的G₂/M期阻滞,并且明显地促进细胞凋亡.     

Effects of Serum Starvation on Radiosensitivity,Proliferation and Apoptosis in Four Human Tumor Cell Lines with Different p53 Status

PURPOSE: The effects of serum starvation on radiation sensitivity, cell proliferation and apoptosis were investigated with particular consideration of the p53 status. MATERIAL AND METHODS: Four human tumor cell lines, Be11 (melanoma, p53 wild-type), MeWo (melanoma, p53 mutant), 4197 (squamous cell carcinoma, p53 wild-type) and 4451 (squamous cell carcinoma, p53 mutant), were used. After the cells had been incubated in starvation medium (0.5% FCS) for 1-6 days, changes in cell cycle distribution, induction of apoptosis and necrosis, and changes in radiation sensitivity were assessed by two-parameter flow cytometric measurements of DNA-dye-exclusion/Annexin V binding, and a conventional colony assay, respectively. RESULTS: p53 wild-type cell lines showed a decrease in the BrdU labeling index and an increase in the apoptotic cell frequency in starvation medium. p53 mutant cell lines showed a decrease in the BrdU labeling index but no evidence of apoptosis. These cells went into necrosis instead. The radiation sensitivity was increased in 4451 and slightly decreased in Be11 and 4197 in starvation medium. CONCLUSION: These data suggest a functional involvement of p53 in starvation-induced G1-block and apoptosis in tumor cells. Altered radiosensitivity after culture in starvation medium seemed to be explained at least in part by the starvation-induced G1-block. The frequency of starvation-induced apoptosis or necrosis was not correlated with radiation sensitivity.     

新辅助化疗的不同方案对进展期大肠癌组织中细胞凋亡和增殖细胞核抗原表达的影响

 目的研究不同新辅助化疗方案能否引起进展期大肠癌肿瘤细胞的凋亡以及对大肠癌肿瘤细胞增殖细胞核抗原(Proliferating cell nuclear antigen,PCNA)表达的影响.方法应用原位末端标记(Terminal deoxynucleotidyl transferase mediated nick endlabeling,TUNEL)和直接免疫荧光法检测细胞凋亡,采用链菌亲和素-生物素-过氧化物酶复合物(Avidin or streptavidin-biotinperoxidase complex,ABC)标记的免疫组化法检测大肠癌组织的PCNA表达,对96例大肠癌患者组织中的正常大肠黏膜和肿瘤细胞的凋亡指数(Apoptotis index,AI)和增殖指数(Proliferative index,PI)分别进行检测.结果两个新辅助化疗组患者肿瘤细胞AI分别为7.47%和6.14%,与手术对照组肿瘤细胞AI 4.83%相比明显增高(P＜0.01),与单纯对照组大肠黏膜细胞AI2.74%相比明显增高(P＜0.01);两个新辅助化疗组中患者肿瘤细胞PI为分别33.71%和39.68%,与手术对照组肿瘤细胞PI51.51%相比明显降低(P＜0.01),与单纯对照组大肠黏膜细胞PI 19.82%相比明显增高(P＜0.01).两个新辅助化疗组间AI与PI相比具有显著差异(P＜0.01).结论新辅助化疗能诱导肿瘤细胞发生凋亡,并能抑制其增殖,细胞增殖与凋亡平衡失调程度与大肠癌新辅助化疗的不同方案密切相关.     

Impact of preirradiation of the tumour bed on cell production rate and cell loss of human FaDu squamous cell carcinoma growing in nude mice.

PURPOSE: To explore whether the tumour bed effect (TBE) in FaDu squamous cell carcinoma growing in nude mice is caused by a reduced tumour cell production rate and/or by increased tumour cell loss. MATERIALS AND METHODS: Human FaDu tumours were studied in NMRI nude mice. The volume doubling time (VDT) between 100 and 400 mm3 was determined for tumours in unirradiated subcutaneous (sc) tissues (group 1), tumours in sc tissues preirradiated with 12.5 Gy (group 2), tumours irradiated in situ with 12.5 Gy (group 3), and tumours from group 3 re-transplanted into unirradiated sc tissues (group 4). Labelling index (LI), potential doubling time (Tpot), relative necrotic area and apoptotic index (AI) were evaluated in tumours from groups 1 and 2. RESULTS: The median VDT were 2.6 days (95% CI 2-4) in group 1 and 7.0 days (4-15) in group 2 (p<0.001). The VDT were not significantly different between groups 2 and 3, and group 1 and 4. In groups 1 and 2, the Tpot values (3.1 +/- 0.6 days (SD) versus 2.9 +/- 0.5 days) and the LI were identical (10 +/- 1.5%). The median relative necrotic area was significantly larger in group 2 (37% [23-42]) compared with group (6% [0.3-27]). The apoptotic index was low (0.2%) and did not differ between groups 1 and 2. CONCLUSIONS: The results indicate that the TBE in FaDu squamous cell carcinoma is not caused by a reduced cell production rate in the viable tumour compartment. Rather, the TBE reflects a decreased viable tumour cell compartment due to increased cell loss. Necrosis appears to be the major component of the tumour bed induced cell loss in FaDu tumours, whereas apoptosis has no impact on the TBE in this model.     

Impact of preirradiation of the tumour bed on cell production rate and cell loss of human FaDu squamous cell carcinoma growing in nude mice

Purpose: To explore whether the tumour bed effect (TBE) in FaDu squamous cell carcinoma growing in nude mice is caused by a reduced tumour cell production rate and/or by increased tumour cell loss. Materials and methods: Human FaDu tumours were studied in NMRI nude mice. The volume doubling time (VDT) between 100 and 400mm3 was determined for tumours in unirradiated subcutaneous (sc) tissues (group 1), tumours in sc tissues preirradiated with 12.5Gy (group 2), tumours irradiated in situ with 12.5Gy (group 3), and tumours from group 3 re-transplanted into unirradiated sc tissues (group 4). Labelling index (LI), potential doubling time (T pot), relative necrotic area and apoptotic index (AI) were evaluated in tumours from groups 1 and 2. Results: The median VDT were 2.6 days (95% CI 2-4) in group 1 and 7.0 days (4-15) in group 2 (p < 0.001). The VDT were not significantly different between groups 2 and 3, and group 1 and 4. In groups 1 and 2, the T pot values (3.1 0.6 days (SD) versus 2.9 0.5 days) and the LI were identical (10 1.5%). The median relative necrotic area was significantly larger in group 2 (37% [23-42]) compared with group (6% [0.3-27]). The apoptotic index was low (0.2%) and did not differ between groups 1 and 2. Conclusions : The results indicate that the TBE in FaDu squamous cell carcinoma is not caused by a reduced cell production rate in the viable tumour compartment. Rather, the TBE reflects a decreased viable tumour cell compartment due to increased cell loss. Necrosis appears to be the major component of the tumour bed induced cell loss in FaDu tumours, whereas apoptosis has no impact on the TBE in this model.     

High levels of chromosome aberrations correlate with impaired in vitro radiation-induced apoptosis and DNA repair in human B-chronic lymphocytic leukaemia cells

Purpose : To investigate the relationship between the susceptibility of B-chronic lymphoid leukaemia (B-CLL) cells to DNA damage-induced apoptosis, the kinetics of DNA strand-break rejoining, and chromosome damage after exposure to ionizing irradiation. Materials and methods : Lymphocytes from B-CLL patients were γ-irradiated in vitro with 0.2-5 Gy and stimulated by Staphylococcus aureus cowan I (SAC I) for estimation of chromosomal damage. Induction of apoptosis after irradiation was studied in 50 patients by two methods: morphological characterization of apoptotic cells after fluorescent staining (Hoechst), and specific quantification of mono- and oligonucleosomes in cytoplasmic cell fractions (ELISA assay). Morphological chromosome damage was scored in the first cell generation after irradiation (13 patients). In parallel, the kinetics of DNA single-strand break rejoining were investigated by the alkaline comet assay (12 patients). Results : Ionizing irradiation did not induce apoptosis in lymphocytes from a subset of B-CLL patients. The results suggest that B-CLL cells resistant to radiation-induced apoptosis could repair DNA strand-breaks more rapidly and showed a higher level of chromosome aberrations than radiation-sensitive B-CLL cells. Conclusion : Each of three biological effects observed (apoptosis, kinetics of DNA single-strand-break repair, chromosomal damage) might be explained by different modifications occurring in irradiated B-CLL cells. Their convergence strongly suggests that resistance to apoptotic death initiation by DNA damage may be impeded by a rapid engaging of the DNA repair mechanisms. The higher level of chromosome aberrations observed in these cells suggests that the type of DNA repair system involved may generate inaccurate repair.     

Radio-induced apoptosis of peripheral blood CD8 T lymphocytes is a novel prognostic factor for survival in cervical carcinoma patients

Background and purpose

A close relationship exists between immune response and tumor behavior. This study aimed to explore the associations between radiation-induced apoptosis (RIA) in peripheral blood lymphocytes (PBL) and clinical pathological variables. Furthermore, it assessed the role of RIA as a prognostic factor for survival in cervical carcinoma patients.

Patients and methods

Between February 1998 and October 2003, 58 consecutive patients with nonmetastatic, localized stage I–II cervical carcinoma who had been treated with radiotherapy (RT) ± chemotherapy were included in this study. Follow-up ended in January 2013. PBL subpopulations were isolated and irradiated with 0, 1, 2 and 8 Gy then incubated for 24, 48 and 72 h. Apoptosis was measured by flow cytometry and the β value, a parameter defining RIA of lymphocytes, was calculated.

Results

Mean follow-up duration was 111.92?±?40.31 months. Patients with lower CD8 T lymphocyte β values were at a higher risk of local relapse: Exp(B) =?5.137, confidence interval (CI) 95?%?=?1.044–25.268, p?=?0.044. Similar results were observed for regional relapse: Exp(B) =?8.008, CI 95?%?=?1.702–37.679, p?=?0.008 and disease relapse: Exp(B) =?6.766, CI 95?%?=?1.889–24.238, p?=?0.003. In multivariate analysis, only the CD8 T lymphocyte β values were found to be of prognostic significance for local disease-free survival (LDFS, p?=?0.049), regional disease-free survival (RDFS, p?=?0.002), metastasis-free survival (MFS, p?=?0.042), disease-free survival (DFS, p?=?0.001) and cause-specific survival (CSS p?=?0.028).

Conclusion

For the first time, RIA in CD8 T lymphocytes was demonstrated to be a predictive factor for survival in cervical carcinoma patients.     

中度高原急性呼吸窘迫综合征的氧缺陷类型

目的：探寻中度高原地区急性呼吸窘迫综合征(ARDS)的氧缺陷类型；方法：对西宁地区海拔(2260m)32例ARDS患者进行血液动力学及血气分析、测定心排指数(CI)、循环阻力(SVRI)、氧输送(DO)、氧消耗(VO)、氧摄取率(ER)以及临床一般资料进行分析；结果：ARDS血液动力学早期表现为高心排指数、低系统阻力，绝大多数表现高氧耗型氧缺陷，随着病情进展，成活组CI、SVRI趋于正常，死亡组则表现持续高动力循环，氧缺陷表现为低氧供型；病程晚期成活组CI、SVRI、DO、VO正常，死亡组仍呈高动力循环；讨论：DO下降，但不伴ER的代偿性增高，出现低氧供型和病理性氧依赖是ARDS的主要死亡原因。