丝裂霉素C诱发大鼠外周血G0期淋巴细胞核损伤指标的比较观察

本文用不同剂量的MMC处理大鼠外周血,放置18小时后直接制备淋巴细胞血涂片,进行观察。实验结果表明,MMC可诱发G_(?)期淋巴细胞的核损伤。本实验对各种核损伤指标进行了比较研究,发现微核、核变形和核碎裂等指标与MMC剂量间有很好的相关性。作者认为,大鼠外周血G_(?)期淋巴细胞核异常测试法比传统的微核测试法更敏感、合理,在致突变试验中可作为初筛的体外短期测试法,值得进一步研究。     

高能电子束超剂量照射与效应关系的探讨——人周血淋巴细胞核损伤指标的比较观察

为了探讨在大剂量辐射条件下建立生物量计的可能,本实验研究了在10MeV电子束照射后,人体外周血淋巴细胞各种核损伤指标的改变与剂量(0—40Gy)间的关系,结果表明,随着照射剂量的增大,复合核损伤指标——核异常率、微核率、核固缩率和核裂解率亦随之上升,在0—20Gy范围内,与剂量呈线性关系,相关系数显著性检验P<0.01的核损伤指标是:核异常率、微核率。P<0.05的是核变形率;在0—40Gy范围内,与之呈线性关系,P<0.01的仅有核固缩率。和染色体畸变分析相比,核异常检测方法简便,可反映超剂鞋(如>10Gy)辐射的损伤,值得引起研究者的注意。     

培养法与直接法外周血淋巴细胞微核率的比较研究

用⁶⁰Co r线1.0-5.0Gy照射狗,比较照后12小时培养法和直接法外周血淋巴细胞微核率,结果微核率随剂量增加而增加,培养法增加的速度和幅度明显高于直接法,经统计学分析二者存在一定的相关性。     

漏出液用于外周血淋巴细胞培养

为研制一种适宜淋巴细胞生长的天然培养基，我们在漏出液中添加不同浓度的小牛血汪有为培养液，对30例外周血淋巴细胞进行常规培养、制片，并以M199作对照，用淋巴细胞转化率、有丝分裂指数、分裂相质量等指标综合评价漏出液的培养效果。结果显示：淋巴细胞在漏出淮中生长快，分裂旺盛，实验组的各项指标均与对照组无差异（P＞0.05）。     

人体外周血淋巴细胞培养及染色体标本制备失败可能原因分析

在开设人类外周血淋巴细胞培养实验中 ,我们采用的实验方法是 :将教师事先配制好的经试培无菌后的 ,含 2 0 %小牛血清的 RPMI1 64 0培养液分装到 2 5m L圆形培养瓶中 ,每瓶 5m L ,每个学生一瓶。实验从抽血→接种→培养→加秋水仙素→收集细胞制片→核型分析等步骤均由学生自己完成。实验的具体操作方法是 :每个实验班 ( 2 0人 )由两个学生 (男女各 1名 )自愿献血 ,静脉取血 4 m L,肝素抗凝 ,每个学生在超净工作台内向培养液内加入一定量血液 ,然后将培养瓶放进 3 7℃恒温箱中培养 72 h,在收集细胞制备染色体之前的 2～ 3h,向培养瓶中加…     

应用大鼠G_0期淋巴细胞核异常测试法评价铬的诱变效应

本文应用大鼠外周血G0期淋巴细胞核异常测试法对铬的诱变效应进行了研究．实验结果表明,不同价太铬具有不同的诱变性。与常用的短期致突变测试法的实验结果相一致,本方法作为环境诱变剂的短期初筛试验有推广应用的价值。     

培养法与直接法外周血淋巴细胞微核率的比较研究

用⁶⁰Co r线1.0-5.0Gy照射狗，比较照后12小时培养法和直接法外周血淋巴细胞微核率，结果微核率随剂量增加而增加，培养法增加的速度和幅度明显高于直接法，经统计学分析二者存在一定的相关性。     

人的外周血淋巴细胞培养及染色体制作技术

从肘静脉采血于含有肝素的一次性注射器中,采用RPMI1640全培养基进行体外淋巴细胞培养。通过对采血后细胞培养时间、秋水仙素浓度及加入时间、低渗时间及培养温度等条件的模索．建立了较成熟的淋巴细胞体外培养方法及染色体制作技术。     

应用大鼠Go期淋巴细胞核异常测试法评价铬的诱变效应

本文应用大鼠外周血G_o期淋巴细胞核异常测试法对铬的诱变效应进行了研究．实验结果表明,不同价态铬具有不同的诱变性．与常用的短期致突变测试法的实验结果相一致,本方法作为环境诱变剂的短期初筛试验有推广应用的价值．     

雷公藤多甙诱发人外周血Go期淋巴细胞微核及核损伤的观察

本实验应用不同剂量的雷公藤多甙处理人外周血Ｇｏ期淋巴细胞,放置１８小时后,直接观察微核率及核损伤的变化。结果显示,雷公藤多甙对人外周血Ｇｏ期淋巴细胞微核率,核变形率和核碎裂率均无明显的影响,亦无明显的剂量依赖性增加。因此,作者认为,雷公藤多甙是一种对人类无潜在遗传毒性的药物,临床上可以安全作用,另一方面,本实验方法比传统的微核测试法更为直接,简便,敏感,有望成为一种简便实用检测化学诱变剂的诱变作用     

单细胞微凝胶电泳技术在人血淋巴细胞DNA损伤研究中的应用

介绍了单细胞微凝胶电泳(SCG)技术的原理和操作过程,并应用该技术研究了γ-线照射、过氧化氢(H_2O_2)、氯化镉(CdCl_2)对人血淋巴细胞DNA的损伤效应。结果表明,γ-线照射、H_2O_2和CdCl_2均能引起细胞DNA迁移长度增加,且呈显著的剂量效应关系。对未处理对照细胞DNA迁移的原因及SCG实验操作过程中应注意的事项也进行了讨论。     

Protective Effect of Carvacrol on Oxidative Stress and Cellular DNA Damage Induced by UVB Irradiation in Human Peripheral Lymphocytes

Exposure to ultraviolet B (UVB; 280‐320 nm) radiation induces the formation of reactive oxygen species (ROS) in the biological system. In this study, we examined the protective effect of carvacrol on UVB‐induced lipid peroxidation and oxidative DNA damage with reference to alterations in cellular an‐tioxidant status in human lymphocytes. A series of in vitro assays (hydroxyl radical, superoxide, nitric oxide, DPPH (2,2‐Diphenyl‐1‐picryl hydrazyl), and ABTS (2,2‐azino‐bis‐3‐ethylbenzothiazoline‐6‐sulfonic acid) radical scavenging assays) demonstrate antioxidant property of carvacrol in our study. UVB exposure significantly increased thiobarbituric acid reactive substances (TBARS), lipid hydroperoxides (LHPs), % tail DNA and tail moment; decreased % cell viability and antioxidant status in UVB‐irradiated lymphocytes. Treatment with carvacrol 30 min prior to UVB‐exposure resulted in a significant decline of TBARS, LHP, % tail DNA, and tail moment and increased % cell viability as carvacrol concentration increased. UVB irradiated lymphocytes with carvacrol alone (at 10 μg/mL) gave no significant change in cell viability, TBARS, LHP, % tail DNA, and tail moment when compared with normal lymphocytes. On the basis of our results, we conclude that carvacrol, a dietary antioxidant, mediates its protective effect through modulation of UVB‐induced ROS.     

Genotoxicity Test of Self-Renovated Ceramics in Primary Human Peripheral Lymphocytes

Zirconia-based ceramics is widely used in dentistry. Different compositions of ceramics have different features. Our self-renovated ceramics become more machinable without scarifying its dental restoration properties after adjusting ratio of lanthanum phosphate (LaPO₄)/yttrium oxide (Y₂O₃). In order to evaluate its safety, here, we tested its genotoxicity in primary human peripheral lymphocytes. The human lymphocytes cultured on three groups of different ratios of LaPO₄/Y₂O₃ diphase ceramics for 6 days showed little effect of growth inhibition and similar effect of growth trend to the negative control. Furthermore, single-cell gel electrophoresis (comet assay) indicated that there was no significant difference of the value of tail moment between the tested ceramics and negative control, the IPS Empress II (P > 0.05). Our findings implicate that our self-renovated ceramics do not induce DNA damages in human peripheral lymphocytes and support their future clinic application.     

X射线对人外周血淋巴细胞DNA损伤修复和凋亡的影响

目的 研究不同时间诱导X射线照射的淋巴细胞进入细胞周期DNA损伤修复与凋亡的影响.方法 X射线（0.5 Gy）作用于正常人外周血淋巴细胞,以照射后不同时间点（0、4 h）分别加入PHA并分成两组,即照射后0 h加PHA组（A组）和照射后4 h加PHA组（B组）,再分别培养0、0.5、2 h,用流式细胞术和免疫印迹法检测A组和B组γ-H2AX蛋白的表达,Annexin-V/PI法分析A、B两组的细胞凋亡率.结果 流式细胞术及免疫印迹结果均显示A组的γ-H2AX蛋白表达高于B组（P＜0.05）,且均先升高后降低.A组细胞凋亡率亦大于B组.结论 不同时间诱导被打击的淋巴细胞进入周期其可能发生DNA修复并同时伴随细胞凋亡的发生.     

HIV-1 Tat蛋白通过增加线粒体ROS产生诱导人外周血B淋巴细胞凋亡

目的:探讨HIV-1 Tat蛋白对人外周血B淋巴细胞增殖、凋亡的影响及其机制。方法:采用流式细胞分选术分离HIV阳性患者外周血单核细胞的B淋巴细胞,分别转染pTat或pcDNA3.1各10μg(分别为pTat组与pcDNA3.1组),采用MTT实验检测细胞胞增殖情况,流式细胞术检测凋亡情况,DCHF-DA测定ROS水平,彗星试验检测细胞DNA损伤情况。结果:pTat组转染24h、48h的细胞增殖抑制率、细胞凋亡率及线粒体ROS水平均显著高于pcDNA3.1组(P0.05)。pcDNA3.1组细胞的DNA大部分呈圆形荧光团,无拖尾现象;pTat组的细胞DNA拖尾现象,呈现典型彗星图像。与pcDNA3.1组相比,pTat组细胞DNA尾长、尾部DNA比例均显著增加(P 0.05)。结论:HIV-1 Tat蛋白可能通过增加线粒体ROS产生,诱导DNA损伤,进而抑制人外周血B淋巴细胞增殖并促进其凋亡。     

Transport of the Immunosuppressant 15-Deoxyspergualin in Human Peripheral Blood Lymphocytes

15-deoxyspergualin (DSG) is a potent immunosuppressive compound currently in clinical trials. In this study, we have characterized the uptake and intracellular localization of DSG in human peripheral blood lymphocytes (PBL′s). DSG is transported into human PBL′s and reaches an estimated maximum concentration of approximately 500μM in 6 hours. The majority of the [³H]-DSG remains in the cytoplasm of cells and that which is associated with the nucleus is only loosely associated. DSG was transported by HeLa cells, as well, suggesting uptake is not specific for hematopoietic cells. Positively charged amino acids and polyamines, which are structurally similar to DSG, were unable to compete for DSG transport suggesting that DSG is transported into cells via a pathway distinct from amino acids or polyamines.     

Modulation of Apoptosis Induced by Tricyclic Antidepressants in Human Peripheral Lymphocytes

We have recently reported that the tricyclic antidepressants (TCAs) imipramine, clomipramine, and citalopram induce apoptosis in human peripheral lymphocytes. This system is well suited for studies on the pathophysiology/physiology of apoptosis regulation. Apoptosis was determined using both DNA gel electrophoresis and flow cytometric analysis. TCA-induced apoptosis in lymphocytes was monitored in the presence of the protein synthesis inhibitor cycloheximide (CHX), the RNA synthesis inhibitor actinomycin D (Act D), the antioxidant reduced glutathione (GSH), the nuclease inhibitor aurintricarboxylic acid (ATA), the cytokine interlukin-2 (IL-2), and the immunostimulator linomide. CHX and Act D failed to prevent and actually enhanced TCA-induced apoptosis in lymphocytes, indicating that protein and RNA syntheses are not required for this process. Exogenous IL-2, GSH, and ATA protected the lymphocytes from apoptosis induced by TCAs in a dose-dependent manner, whereas linomide had no effect on TCA-induced apoptosis under our in vitro conditions. Our data demonstrate that TCA-induced apoptosis in human lymphocytes shares many common features with other stimuli-induced apoptotic processes. © 1997 John Wiley & Sons, Inc. J Biochem Toxicol 12: 115–123, 1998     

抗癌药对黑胸大蠊淋巴细胞和生殖细胞核结构的损伤

噻替派浓度为0.1%、0.3%、0.5%时,黑胸大蠊精母细胞染色体断裂和裂隙率分别为6.3%、 10.5%和14.2%,显著地高于对卵母细胞的影响;和雄虫外周血淋巴细胞微核率呈平行关系,随微核率增多而增加。5-氟尿嘧啶浓度为0.1%、0.3%和0.5%时,卵母细胞染色体断裂和裂隙率分别为3.5%、9.8%和16.2%,和雌虫外周血淋巴细胞微核率呈平行关系,随微核率增多而增加,而对雄虫生殖细胞影响不显著。 Abstract:0.1%,0.3%,0.5% Thio-TEPA induced 6.3%,10.5% and 14.2% chromosome break or gap in spermatocyte of cockroach respectively.This was markedly higher than those in oocyte.In doses from 0.1 to 0.5 Tho-TEPA the frequency of micronucleus increased parallely with nuclear damage.0.1%,0.3%,0.5% 5-fluorouracil induced 3.5%,9.8%,16.2% chromosome break or gap in oocytes respectively.This was paralled with the frequency of micronucleus in lymphocytes of the female.5-fluorouracil showed not marked effect on spermatocyte.