HepG2细胞染色体着丝粒点变异的研究

背景与目的:探讨HepG2瘤细胞染色体着丝粒点(Cd)的变异与其非整倍性畸变的关系.材料与方法:用Cd-NOR同步银染分析技术研究HepG2细胞染色体Cd的变异.结果:HepG2细胞染色体Cd缺失率为2.30%、Cd迟滞复制率为1.02%、小Cd率为2.58%、Cd-NOR融合率为0.64%;与正常人胚胎绒毛细胞染色体Cd变异相比较,HepG2细胞染色体Cd缺失、Cd迟滞复制和小Cd升高,而Cd-NOR融合差异无显著性.结论:HepG2细胞染色体非整倍性畸变的途径可能主要涉及Cd缺失、Cd迟滞复制、小Cd.     

Retrospective analysis of clonality and detection of residual disease in myeloid leukemia by FISH on long-term stored bone marrow smears

Abstract Background. Fluorescence in situ hybridization (FISH) has allowed the detection of numerical chromosomal aberrations in interphase nuclei on fresh or frozen smears of leukemia.
Methods. To analyze clonality and residual disease in myeloid leukemia retrospectively, we applied FISH to bone marrow smears stored at ambient temperature for up to 9 years.
Results: When hybridization efficiency was investigated on stored control smears from patients without hematological malignancy, more than 96% of nuclei showed the expected number of signals using DNA probes specific for chromosome 7, X or Y. In combination with cell morphology, we observed much higher hybridization efficiency in blasts and granulomonocytic cells compared with lymphoid and erythroid cells. On the basis of good hybridization efficiency for old smear specimens, we applied FISH to stored bone marrow smears of myeloid leukemias, in which either loss of chromosome 7 or loss of sex chromosomes had been verified previously by conventional cytogenetics (one patient with chronic myelomonocytic leukemia (CMML) and four with acute myeloid leukemia (AML; three M2 and one M7)). As a result, the loss of chromosome was detected in blasts from all patients and was observed in mature granulocytes, except in M7. In the CMML patient and one AML (M2) patient with t(8;21), lymphoid and erythroid cells also showed the loss of chromosomes, suggesting that it should occur at stem-cell level. A high amount of residual disease was detected in the morphological remission samples in one AML (M2) patient after induction therapy. The patient eventually succumbed to relapse.
Conclusion Thus, the present FISH technique is useful to analyze the clinical significance of clonality and the residual disease in myeloid leukemia, retrospectively.     

Absence of ATM truncations in patients with severe acute radiation reactions

Purpose: Severe acute toxicity limits the effective use of radiotherapy in patients who are radiosensitive, and it is not usually possible to identify these radiohypersensitive (R-H) individuals before treatment commences. Five such R-H patients were detected over a 3-year period. We undertook this study to determine whether the severe acute radiohypersensitivity of these five individuals showed any correlation with cellular and molecular parameters known to be abnormal in radiosensitivity-related syndromes such as ataxia–telangiectasia (A-T).

Methods and Materials: Lymphoblastoid cells were isolated from fresh blood from the 5 R-H individuals who had previously demonstrated clinical R-H at least 9 months prior to sampling. Lymphoblastoid cell lines (LCLs) were established to determine the extent of postradiation chromosomal aberrations, cell cycle delay, cell proliferation, and tumor suppressor p53 protein stabilization. The polymerase chain reaction (PCR) and protein truncation (PTT) assays were used to test for the possibility of mutations in the gene mutated in A-T, termed ATM.

Results: LCLs derived from R-H subjects retained a significantly higher degree of radiation-induced chromosomal aberrations when compared to normal control LCLs. p53 stabilization by ionizing radiation appeared normal in all but one R-H subject. There was no evidence of A-T gene truncation mutations in any of the R-H subjects tested.

Conclusions: All R-H subjects in this study had their cellular radiosensitivity confirmed by the chromosomal aberration assay. Delayed p53 stabilization at 4 hours postirradiation in one R-H subject suggested that different etiologies may apply in the radiohypersensitivity investigated in this study.     

丹参对丝裂霉素C诱发小鼠生殖细胞遗传损伤的防护作用研究

本文根据动物实验模型的设计，研究了中药丹参对丝裂霉素Ｃ（ＭＭＣ）诱发雄性小鼠过生殖细胞遗传损伤的防护效应。雄性昆明小鼠随机分成８组，每组１５只，实验组分别注射高、中、低剂量丹参和ＭＭＣ，观察动物的精子畸形率、早期精细胞微核率和精原细胞染色体畸变率。结果表明，中药本身无诱变损伤作用，而对ＭＭＣ有拮抗抑制作用。提出中药丹参具有抗ＭＭＣ诱发小鼠生殖细胞遗传损伤的作用。     

氟吗啉诱发V79和CHL细胞基因突变和染色体畸变的研究

目的 探讨氟吗啉的致突变性。方法 首先测定氟吗啉对V79和CHL的细胞毒性，然后在非代谢活化和代谢活化条件下，进行氟吗啉诱发V79细胞HGPRT基因突变试验和CHL细胞染色体畸变试验。结果 以氟吗啉500、100、20和4μg/mL浓度处理V79细胞，处理组诱变率与阴性对照组突变率比较，差异无显著性(P≥0．05)；以500、250、125和62．5μg／mL浓度处理CHL细胞24、48h后，在非代谢活化条件下，处理组染色体畸变均小于5％，但在代谢活化条件下，处理组染色体畸变率均大于5％，而且呈剂量．反应关系，通过G-显带发现断裂集中发生于4q上，是断裂热点。结论 可以认为应用氟吗啉100～200mg／L防治植物病害不会对人类健康造成危害，但是职业人群应注意防护。     

守宫木细胞与遗传毒性实验研究

目的：以体外试验研究野生蔬菜守宫木的细胞毒性和遗传毒性。方法：生（Ⅰ）和煮熟（Ⅱ）的守宫木匀浆液分别以40000、20000、10000、5000、2500、1250μg／ml浓度对CHL细胞染毒，每一浓度分别在24、48和72h时间点以中性红摄入方法对守宫木的细胞毒性进行评定；体外染色体畸变试验则以同样的浓度分别以生和煮熟的守宫木匀浆液染毒2h后，收获细胞制备中期相，对染色体结构畸变进行分析。结果：中性红摄入细胞毒性试验：在生的与煮熟的守宫木匀浆液细胞毒性剂量效应关系分析中，低剂量组随着染毒浓度加大细胞毒性增强，而在较高剂量组则出现随染毒浓度加大细胞毒性减弱，20000和40000μg／ml剂量组出现细胞的增殖现象；时间效应分析中有和剂量效应分析相似的现象，即在低剂量组随着染毒时间的延长细胞毒性增强，而从10000μg/ml剂量组开始随着染度时间的延长细胞毒性减弱并出现细胞的增殖现象；对生的与煮熟的守宫木细胞毒性进行比较分析，仅在染毒24h的20000μg／ml剂量组发现煮熟的守宫木匀浆液的细胞毒性有统计学意义（P〈0．05）地高于生的守宫木匀浆液。体外染色体畸变试验结果阴性。结论：守宫木在一定浓度范围内对细胞溶酶体和高尔基体有一定的损伤作用，未观察到守宫木对细胞的遗传物质染色体有损害作用，未发现不同的食用习惯对守宫木的毒性有影响。     

子宫内膜癌染色体不稳定性研究

目的探讨子宫内膜癌染色体不稳定性。方法取21例子宫内膜癌患者外周血淋巴细胞体外常规培养、制片、GTG显带分析染色体畸变(CAR),Brdu法分化染色分析姊妹染色单体交换(SCE)、微核(MN)、核仁组织形成区(NOR)。结果内膜癌组(21例)染色体结构畸变以出现次数的多少排列为:1、3、2、5、7、8号,染色体数目、结构畸变率分别为12.86%6、.1%,SCE频率、MN率、Ag-NOR分别为7.4±1.5次/细胞(、6.6±1.9)‰、7.1±1.6个/细胞,对照组(10例)染色体数目、结构畸变率分别为3.4%、0.4%,SCE频率、MN率、Ag-NOR分别为4.8±0.6次/细胞(、2.0±0.9)‰、4.9±1.1个/细胞,内膜癌组均明显高于对照组(P<0.01)。结论子宫内膜癌病人外周血淋巴细胞染色体数目和结构畸变率增高。     

苯作业工人外周血淋巴细胞染色体畸变分析

目的 研究苯接触工人外周血淋巴细胞染色体畸变的水平,以及接苯工人的工种、工龄等对结构畸变细胞率的影响.方法 选取130名接触苯作业工人进行基本情况调查.并抽取每人外周静脉血0.5 ml,加入含有植物血凝素和小牛血清的1640培养基中培养52～54 h,终止培养4～6 h前加入秋水仙素.收获细胞,染色,用自来水、单蒸水冲洗,干后封片光学显微镜下观察.以非分带染色的方法检测外周血淋巴细胞染色体畸变种类和异常率.结果 苯接触工人的染色体畸变异常人数在包括裂隙时有40人,在不包括裂隙时有22人,分别占总人数的31.7%与16.9%.在分析不同工种、性别、工龄、饮酒、吸烟、防护知识的掌握程度不同对染色体畸变的影响时,无论包括不包括裂隙这种畸变种类,工种是其影响因素.染色体结构畸变类型主要是单体裂隙31.77%、单体缺失30.75%和断裂25.41%.没有发现上龄、性别、饮酒、吸烟和防护知识对染色体畸变率有明显影响.结论 苯接触工人的染色体异常率在包括裂隙时为31.7%,在不包括裂隙时为16.9%.油漆工人的染色体畸变异常率高于其他苯作业工人.     

不同程度近视患者行SMILE术后的有效光学区及角膜高阶像差比较

目的：比较低、中、高度近视患者行飞秒激光小切口基质透镜取出术(SMILE)后的有效光学区和角膜高阶像差。

方法：收集2019-02/2021-02在我院行SMILE手术的患者134例,均取右眼入组,按等效球镜度(SE)分为低度近视组SE>-3.00 D,中度近视组-6.00 D结果：SMILE术后1 mo,三组术后有效光学区均小于预设光学区,随着屈光度数增加,有效光学区越小(P<0.05); 术后1 mo角膜总高阶像差、球差、彗差均高于术前,除低近视度组球差术前与术后1 mo无差异(P>0.05),其余组别均有差异(P<0.05); 角膜总高阶像差、球差和彗差随着屈光度数增加而增加,三组角膜总高阶像差和球差有差异(均P<0.05),高度近视组与中度近视组、低度近视组彗差结果均有差异(P<0.05),中度近视组与低度近视组彗差结果无差异(P>0.05)。

结论：随着手术矫正的屈光度数增加,术后有效光学区减少越多,角膜高阶像差增加越明显; SMILE术后1 mo角膜高阶像差较术前增加。     

试色糊剂对Ips-empressⅡ全瓷冠颜色的影响

目的：观察试色糊剂对全瓷冠颜色的影响。方法：在临床上收集缺损、变色或过小的上前牙共计50颗,行全瓷冠修复（Ips-empressⅡ全瓷系统制作）。粘结前由同一实验人员用试色糊剂试色,将全瓷冠就位未涂试色糊剂时作为对照组,用电脑比色仪测定对照组及5种试色糊剂[A1、A3、Transluent（Trt）、Whiteopaque（WOT）和B0.5opaque（B0.5）]试色后的L、a、b值,用卡尺测量被测点的瓷层厚度,根据公式色ΔEab=（ΔL^2＋Δa^2＋Δb^2）1/2、Lab=L、Cab=（a^2＋b^2）^1/2、Hab=arctan（b/a）计算并分析每种糊剂试色后的色差、明度、饱和度和色相角。结果：50颗全瓷冠被测点（唇面中1/3）的平均瓷层厚度为1.668mm,用5种试色糊剂试色后的Lab、Cab、Hab值与对照组相比差异无显著性（P〉0.05）;5种试色糊剂引起的色差均小于1.5NBS,临床上肉眼均难以察觉。结论：当全瓷冠的瓷层厚度达到1.7mm时,粘结剂对修复体颜色的影响较小,肉眼难以察觉。随着瓷层厚度的下降,应考虑树脂粘结剂的遮色作用。