Influence of Treatment Time on the Response of Rat Rhabdomyosarcoma Rlh to Fractionated Irradiation

The aim of this work was to study influence of overall treatment time on response of an experimental tumour to fractionated radiotherapy. Rhabdomyosarcoma RlH of the rat was treated by fractionated irradiation applying 30 fractions in different overall treatment times of 10, 18.5, 29, 39, and 67 days. The doses per fraction ranged from 1.50 to 2.67 Gy. Tumour response was assessed by net growth delay and by an in vitro colony assay. The results indicate that repopulation of the tumour with clonogenic tumour cells slowed down during treatment leading to an average doubling time of clonogenic tumour cells of 8 ± 1 days, which is considerably longer than that of unirradiated tumours (3 days). The repopulation rate seemed to increase after about 3 weeks of treatment with a doubling time of tumour clonogens of 5 ± I days, which was still lower than in control tumours.     

六种小鼠肿瘤及白血病细胞株的细胞动力学研究

本文对L7811-85,L7212-85,L1210-86,P388-86 4个淋巴细胞白血病及S180-86和肝Ca-86肿瘤细胞株分别测定了细胞生长曲线、克隆形成效率和细胞DNA含量分布等参数。对其中L7811-85细胞株进行了较系统的细胞动力学研究,并应用~3H-TdR自杀试验微培养测定其克隆形成细胞的增殖时和合成时。结果表明:这6种细胞株均可在体外无刺激因子作用下良好生长。并可见随细胞传代数增加,细胞增殖加速,克隆形成率提高。用~3H-TdR自杀试验测出的克隆形成细胞增殖时与群体细胞倍增时相比,可反映其潜在倍增时。     

Study on Fractionated Total Body Irradiation before Hematopoietic Stem Cell Transplantation

OBJECTIVE To observe the dose and the complications from total body irradiation before hematopoietic stem cell transplantation.METHODS This study involved 312 patients with total body irradiation before hematopoietic stem cell transplantation. They were entered into the treated research from May 1999 to October 2005. All patients had Received the irradiation from 60Co of an absorbed dose rate of (5.2 ± 1.13) cGy/min. The total dose of TBI was 7~12 Gy, 1 f/d × 2 d. A high-dose rate group (≥ 10 Gy) included 139 cases and a low-dose rate group (< 10 Gy) included 173 cases.RESULTS The probability of acute gastrointestinal reactions in the high-dose rate group was more compared with that in the low-dose rate group. The differences for other reactions, such as hematopoietic reconstitution and graft survival rate, between the two groups were insignificant.CONCLUSION Using fractional total body irradiation at a dose rate of 5 cGy/min, with a total dose of 7~12 Gy, 1 f/d x 2 d, with the lung receiving under 7.5 Gy is a safe and effective pretreatment for hematopoietic stem cell transplantation.     

Effect of hyperthermia on cyclin B expression in a human glioblastoma cell line

Summary Effects of hyperthermia on the cell kinetics of glioblastoma cells were investigated using flow cytometry. Pulse-labeling with 5-bromodeoxyuridine (BUdR) and chasing of the labeled cells revealed temporary accumulation of the labeled cells in G₂M phase and a reduction of DNA synthesis.The level of cyclin B rises rapidly in G₂ phase and falls at the end of mitosis in normal cycling cells. Cyclin B binds to p34^cdc2, resulting in histone kinase activity which is necessary for the initiation of mitosis. The amount of p34^cdc2 remains constant throughout the cell cycle. The level of cyclin B was measured using an anti-cyclin B antibody and flow cytometry in order to investigate the cause of the G₂ accumulation induced by hyperthermia. A low level of cyclin B, in comparison with that of normal cycling cells, persisted for more than 3 h after hyperthermia. These results indicate that the temporary accumulation of cells in G₂M phase after hyperthermia may be caused, at least in part, by an insufficient level of cyclin B.     

Early Effects of Preoperative Irradiation Upon the Cell Cycle Composition in Rectal Adenocarcinomas a Flow-Cytometric DNA Investigation

The DNA patterns were studied by means of flow cytometric analysis in 43 rectal adenocarcinomas. Ploidy level and cell cycle distribution were related to clinical stage and histopathology. The frequency of grossly aneuploid tumours and tumours with multiple aneuploid cell populations increased with more advanced clinical stages and with the degree of dedifferentiation. In 15 cases the DNA pattern was studied before and after preoperative irradiation. The ploidy level was not affected by irradiation. A pronounced increase in the proportion of G₂ cells was found after irradiation. This G₂ blockage was proportional to the amount of S-phase cells before irradiation. Since following irradiation the proportion of S-phase cells was low and the proportion of G₁ cells unchanged, the existence of a high fraction of resting G₁ tumour cells can be assumed.     

Effect of 5-Fluorouracil on the Cell Growth and Cell Cycle Kinetics of a Mouse Ascites Tumor Growing in Vivo

The effect of 12, 24 and 36 mg/kg body weight doses of fluorouracil (5-FU) on the Bp8 ascites sarcoma growing in vivo was studied. From sequential studies of the total number of cells together with the composition of cells in the cell cycle, the cell cycle flow was calculated and correlated to the pharmacokinetics, which was determined by using ³H-5-FU. The dose of 12 mg/kg 5-FU affected cell growth between 24 and 72 hours, while the effect of higher doses was immediate. An early block in outflow of cells from G₁ was followed by an increased outflow, indicating an early inhibition followed by an enhancement of the initiation of the DNA synthesis. This increased outflow from G₁ together with the decrease in outflow from the early S-phase, i.e. decreased DNA synthesis, resulted in an accumulation of cells in the early part of the S-phase. The prolonged effects on the cell growth and the cell cycle flow despite the very fast decline in the drug concentration both in the ascites fluid and within the cells, together with a constant level of the drug in the macromolecular fraction, suggest an interaction between 5-FU and RNA/DNA at later times rather than an inhibition of the thymidylate synthetase activity.     

薏苡仁酯对人鼻咽癌细胞辐射效应的协同作用

 目的 探讨薏苡仁酯(coixenolide,CXL)对人鼻咽癌细胞CNE-2Z辐射效应的影响.方法 以60Co为放射源,采用微量细胞克隆形成法检测CNE-2Z对γ射线的敏感性.结果 CXL使CNE-2Z的辐射-存活曲线向左移,Do、Dq和N值下降.不同浓度的CXL(5-20 μmol/L)使放射剂量减少17.26%-41.90%(在D37水平),其增效比(ER):Do比值1.14-1.64,Dq比值1.27-1.79.另外,CXL和射线的合并效应大于两者单独效应之和.结论 CXL和射线的相互作用产生协同效应.     

Cell kinetic analysis of non-Hodgkin's lymphomas using in vivo iododeoxyuridine incorporation and flow cytometry

The aim of this study was to analyse dynamic cell proliferation parameters in non-Hodgkin's lymphomas. Sixty-one patients with newly diagnosed or with recurrent disease were given iododeoxyuridine (IdUrd) intravenously near 4 h prior to tumour biopsy. After staining with an IdUrd reactive antibody and propidium iodide, S-phase fraction (SPF), labelling index (LI), S-phase duration time (Ts) and potential tumour doubling time (Tpot) were determined by flow cytometry. Thirty-eight samples, 15 low grade (LGM) and 23 high grade (HGM) malignant lymphomas, were possible to evaluate. Twenty-three cases were excluded due to aneuploidy, insufficient amount of material or technical problems. Tpot values varied between 0.8–32.9 days (mean 7.0 days). HGM lymphomas had shorter mean Tpot times than LGM lymphomas (4.8 versus 10.4 days, p=0.05). For Ts the range was 4.2–20.1 h (mean 9.1 h), and a difference between the two histological groups was demonstrated with a longer mean Ts for HGM compared with LGM cases (10.0 versus 7.8 h, p=0.04). Tpot showed a negative correlation with SPF (P=0.003), and Ts demonstrated a positive correlation to SPF (p=0.02). The clinical significance of the dynamic cell proliferation parameters studied remains to be clarified, but the interelationships between Ts/SPF and Ts/morphologic subtype might be factors of interest for future prognostic studies in malignant lymphomas.     

Effect of Hemopoietic Growth Factors on the Proliferation of Acute Myeloid and Lymphoid Leukemias

The effect of G-CSF, GM-CSF, IL-1 and IL-3 on the proliferation of acute leukemia cells (evaluated as ³HTdR uptake) was investigated in short-term liquid cultures and compared with that observed on normal bone marrow (BM) populations enriched for immature cells. In acute myeloid leukemias (AML), a marked leukemic proliferation was induced in 10/18 cases by IL-3, in 9/18 by GM-CSF, in 7/18 by IL-1 and in 4/18 by G-CSF. In acute lymphoid leukemias (ALL), marked stimulation was observed in 7/11 cases with IL-3 and in 5/11 with GM-CSF, whereas IL-1 and G-CSF were ineffective. Both in AML and ALL, the combination of several factors did not result in an additive synergistic effect. Purified normal BM cells responded to all four growth factors and their combinations produced an additive effect on cell proliferation which probably relates to the heterogeneity of the cell populations studied. The effect of a G-CSF, GM-CSF, IL-1 and IL-3 on the proliferation of acute leukemia cells by different growth factors suggests that caution should be exercised in their clinical use in these diseases     

Differential S-phase progression after irradiation of p53 functional versus non-functional tumour cells

Background

Many pathways seem to be involved in the regulation of the intra-S-phase checkpoint after exposure to ionizing radiation, but the role of p53 has proven to be rather elusive. Here we have a closer look at the progression of irradiated cells through S-phase in dependence of their p53 status.

Materials and methods.

Three pairs of tumour cell lines were used, each consisting of one p53 functional and one p53 non-functional line. Cells were labelled with bromodeoxyuridine(BrdU) immediately after irradiation, they were then incubated in label-free medium, and at different times afterwards their position within the S-phase was determined by means of flow cytometry.

Results

While in the p53 deficient cells progression through S-phase was slowed significantly over at least a few hours, it was halted for just about an hour in the p53 proficient cells and then proceeded without further delay or even at a slightly accelerated pace.

Conclusions

It is clear from the experiments presented here that p53 does play a role for the progress of cells through the S-phase after X-ray exposure, but the exact mechanisms by which replicon initiation and elongation is controlled in irradiated cells remain to be elucidated.     

"增免方"对皮肤鳞状细胞癌术后患者免疫功能的影响

目的 探讨"增免方"对皮肤鳞状细胞癌术后患者免疫功能的影响.方法 54例皮肤鳞状细胞癌术后患者给予口服"增免方"治疗,应用流式细胞仪检测治疗前后外周血中CD3+、CD4+、CD8+、CD4+/CD8+、NK细胞等5项免疫指标的变化.结果 患者治疗后CD3+、CD4+、CD4+/CD8+、NK细胞4项免疫指标均高于治疗前(P均<0.01).结论 "增免方"可显著改善皮肤鳞状细胞癌术后患者的免疫功能.     

溶血磷脂酰胆碱对MG-63细胞粘附和迁移能力的影响

目的 探讨溶血磷脂酰胆碱(lysoph-osphatidylcholine,LOA)对骨肉瘤细胞粘附和体外迁移能力的影响.方法 培养骨肉瘤细胞Mg-63,待其生长至对数生长期,传至第3代收集肿瘤细胞,各细胞分为两份.通过LOA预处理细胞后,一份以MTT微量酶比色反应法定量粘附细胞数,一份以划痕损伤实验检测迁移能力.统计分析比较LOA对骨肉瘤细胞迁移和粘附能力的相关性.结果 MTT法检测结果显示,不同作用时间(0、6、12、24、48 h)不同剂量LOA(0.000、0.001、0.010、0.060、0.100 mg/ml)干预后骨肉瘤细胞体外粘附能力明显加强,与对照组(0.000 mg/ml各组)相比,差异有统计学意义;Transwell结果显示,不同作用时间(0、6、12、24、48 h)不同剂量LOA(0.000、0.001、0.010、0.060、0.100 mg/ml)干预后骨肉瘤细胞体外迁移能力随着时间及剂量的增加而明显加强,与对照组(0.000 mg/ml各组)相比,差异有统计学意义.结论 LOA具有促细胞骨架变化进而调节细胞粘附和迁移能力的作用,表明LOA在恶性肿瘤发生进展中可能与肿瘤的侵袭和转移密切相关.     

硝酸羟胺对雄性小鼠生殖细胞和精子影响的初步研究

背景与目的:探讨不同剂量的硝酸羟胺对小鼠睾丸生殖细胞微核、精子畸形以及睾丸生殖细胞DNA的影响.材料与方法:小鼠腹腔注射不同浓度的硝酸羟胺溶液,小鼠生殖细胞微核试验连续4 d染毒,第5 d取双侧睾丸;小鼠精子畸形试验、睾丸生殖细胞DNA流式细胞仪分析连续5 d染毒,染毒后28 d取双侧附睾、睾丸,计数小鼠精子畸形率及生殖细胞DNA分布情况,评价硝酸羟胺对小鼠的生殖毒性作用.结果:腹腔注射硝酸羟胺实验组生殖细胞微核率、精子畸形率与对照组相比差异均有显著性;睾丸生殖细胞流式仪分析中,在高剂量组圆形、长形精子细胞DNA百分率与对照组相比差异有显著性.结论:小鼠腹腔注射硝酸羟胺对雄性小鼠生殖细胞和精子有损害作用.     

累及野调强放疗联合同步化疗治疗局部晚期非小细胞肺癌的疗效观察

目的比较累及野照射（IFI）和选择性淋巴结照射（ENI）调强放射治疗联合同步化疗治疗局部晚期非小细胞肺癌（LA-NSCLC）的不良反应和疗效。方法49例LA-NSCLC患者前瞻性随机分为IFI组和ENI组,同步化疗两周期,行根治性调强放射治疗。结果IFI组和ENI组≥2级放射性肺炎发生率分别为8.0％和37.5％（P=0.01）;GTV平均剂量分别是（66.2±6.5）Gy和（61.3±6.3）Gy,（P=0.01）;总有效率为92.0％和66.7％（P=0.03）;预防照射区内淋巴结复发率为4.2％和4.0％,（P=0.49）;1年局部失败率分别为8.0％和16.7％（P=0.62）;1年生存率为72.0％和62.5％（P=0.48）。结论IFI同步放化疗治疗LA-NSCLC可降低正常组织并发症的概率,提高靶区照射剂量和肿瘤控制率,预防照射区内淋巴结复发率无增加,有望延长患者生存期。     

互隔交链孢霉提取物对人羊膜FL细胞周期影响的研究

本文应用流式细胞光度术(F C M)研究了从食管癌高发区粮食中分离的互隔交链(?)霉提取物对人羊膜 F L 细胞周期的影响。实验结果表明:提取物对指数生长的 F L 细胞的生长育抑制作用并有剂效关系。它能使细胞从 S→G_2的进行受阻,因而 S 期细胞在细胞周期中出现堆积。对细胞的有丝分裂也有一定干扰作用、但不能阻断细胞周期的进行。提取物处理后的细胞,DNA 含量大于4C 的细胞明显增多。这些高倍体细胞多数为非整倍体细胞。     

Cell cycle effects of gemcitabine.

Gemcitabine (2',2'-difluoro-2'-deoxycytidine, or dFdC) is a promising anticancer agent with demonstrated clinical activity in solid tumours currently undergoing clinical trials. Despite extensive studies on the biochemical mechanism of action, cell cycle perturbations induced by dFdC have not yet been thoroughly investigated, apart from the expected inhibition of DNA synthesis. The aim of our study was to clarify whether cell population kinetics is a vital factor in the cytotoxicity of dFdC in single or repeated treatments and in the dFdC-cisplatin combination. Ovarian cancer cells growing in vitro were treated with dFdC for 1 hr in a range of concentrations from 10 nM to 10 microM. Cell kinetics was investigated by DNA-bromodeoxyuridine flow cytometry, using different experimental protocols to measure either the time course of DNA-synthesis inhibition or the fate of cells in G(1), S or G(2)M at the time of dFdC treatment or 24 hr later. A modified sulforhodamine B test was used to assess the growth inhibition caused by dFdC given alone or with cisplatin. Although dFdC promptly inhibited DNA synthesis, cytotoxicity on proliferating cells was not specific for cells initially in the S phase. DNA synthesis was restored after a G(1) block of variable, dose-dependent length, but recycling cells were intercepted at the subsequent checkpoints, resulting in delays in the G(2)M and G(1) phases. The activity of repeated treatment with dFdC + dFdC or dFdC + cisplatin was highly dependent on the interval length between them. These results suggest that the kinetics of cell recycling from a first dFdC treatment strongly affects the outcome of a second treatment with either dFdC itself or cisplatin.     

子宫颈癌外周血T淋巴细胞亚群检测及其临床意义

[目的]探讨子宫颈癌患者细胞免疫功能的变化及其临床意义.[方法]应用间接免疫荧光染色法检测40例子宫颈癌患者外周血T淋巴细胞亚群的分布及天然杀伤细胞(NK细胞)活性.『结果]子宫颈癌患者外周血T淋巴细胞中的CD3 、CD4 比正常人显著降低(P＜0.001),CD8 比正常人显著升高(P＜0.001),CD4 /CD8 比值比正常人显著降低(P＜0.001).老龄子宫颈癌CD4 下降较年轻病例明显(P＜0.05).子宫颈深层浸润病例CD8 上升,NK细胞百分率下降(P＜0.05).免疫功能抑制与病理类型、分化等其他病理因素无关.[结论]子宫颈癌患者的细胞免疫功能严重受阻,CD8 和NK指标可作为浸润性子宫颈癌的指标.     

晚期恶性肿瘤患者化疗前后T细胞亚群变化的临床分析

目的 探讨恶性肿瘤患者化疗前后T细胞亚群的变化.方法采用流式细胞术分别对26例晚期恶性肿瘤患者化疗前和化疗后外周血T细胞及亚群进行检测.结果晚期恶性肿瘤患者化疗前后CD3+T细胞百分数分别为(71.35+ 16.82)%、(70.49+16.82)%,差异无统计学意义(P＞0.05);CD3+ CD4+比值分别为(31...     

鼻咽癌患者治疗中CD34和T细胞亚群检测的意义

[目的]探讨鼻咽癌患者治疗过程中T细胞亚群、CD34检测的临床意义.[方法]应用流式细胞仪测定67例鼻咽癌患者治疗前、治疗4周以及治疗后1个月T细胞亚群、CD34和常规检查血液学情况,20例健康人群为对照组.[结果]血液学常规显示鼻咽癌患者白细胞、血红蛋白、血小板以及尿酸在治疗过程中与对照组比较均无显著性差异(P＞0.05).治疗前CD8+和CD4+/CD8+显著性低于正常组(P＜0.05).患者放疗和同期放化疗期间CD34明显下降,CD3+、CD4+、CD8+、CD4+/CD8+升高.鼻咽癌组治疗后1个月CD8+、CD4+/CD8+显著性高于对照组(P＜0.05).鼻咽癌同期放化疗患者CD34治疗4周和治疗后1个月均显著低于单纯放疗组(P＜0.05),且治疗后1个月CD3+、CD4+显著高于单纯放疗组(P＜0.05).[结论]T细胞亚群变化可评估鼻咽癌患者治疗过程中的免疫功能状态,CD34可用于观察放化疗对骨髓功能影响.     

溶瘤腺病毒Ad-MK联合外照射对膀胱癌细胞增殖的影响

目的探讨以Midkine(MK)基因为启动子的溶瘤腺病毒Ad-MK单独及联合外照射对人膀胱移行细胞癌EJ细胞增殖的影响。方法应用RT-PCR技术检测EJ细胞中MK mRNA的表达及感染Ad-MK后细胞中腺病毒E1a mRNA的表达。细胞存活率检测Ad-MK对EJ细胞放射敏感度的影响。测定病毒复制量观察外照射对Ad-MK复制的影响。结果EJ细胞中可见MK mRNA的表达。Ad-MK感染细胞后可检测到细胞中E1a mRNA的表达。联合组细胞的存活率与Ad-MK或外照射组的差异有统计学意义(P<0.01)。低剂量外照射联合病毒治疗后并未减少病毒在细胞中的复制。结论膀胱移行细胞癌EJ细胞中可见MK mRNA的表达;Ad-MK可在细胞中复制,其对细胞的增殖抑制作用呈量效和时效关系。小剂量外照射能增强Ad-MK的细胞毒作用,并能增加病毒在细胞中的复制。