DNA倍体分析在恶性腹腔积液诊断中的价值

目的探讨DNA异倍体对于恶性腹腔积液的诊断价值。方法抽取64例中等量以上腹腔积液患者的腹水，分离其中的细胞，制备单细胞悬液，应用流式细胞仪分析DNA异倍体。结果DNA异倍体在肿瘤细胞中出现率（82．7％）显著高于非肿瘤细胞（0），统计学检验表明两者有显著性差异。结论流式细胞术分析腹水细胞的DNA异倍体，对于恶性肿瘤的细胞学诊断有较大的意义，可在临床推广应用。     

DNA倍体测定在判断肺癌恶性程度中的作用

了解肺癌细胞核ＤＮＡ倍性分布与肺癌临床病理类型的关系,研究测定癌旁细胞核ＤＮＡ倍性在判断肿瘤局部转移中的作用。方法应用流式细胞仪测定手术切除的４８例新鲜肺癌组织标本,包括１６例癌旁肺组织细胞核ＤＮＡ倍体模式。     

DNA含量测定对恶性腹水的诊断价值

判断腹水性质是临床上一个重要而又棘手的问题,寻找并建立快速、准确判断腹水性质的检测方法和指标在临床上具有重要意义.本实验采用流式细胞仪(Flow Cytometry,FCM)对 25例腹水患者的细胞进行了 DNA含量测定,井对其诊断恶性腹水的价值与传统的细胞形态学方法、腹水癌胚抗原(CEA) 和癌抗原 19-9(CA19-9) 进行了比较和分析. 1 材料与方法 1.1 对象 健康正常成人 10名.腹水患者 25例,男 16例,女 9例,年龄 18岁 -81岁,中位年龄 56岁.恶性腹水 12例,其中胃癌腹膜转移 6例,原发性肝癌 3例,结肠癌腹膜转移 2例,卵巢癌腹膜转移 1例 ; 良性腹水 13例,其中结核性腹膜炎 6例,肝硬化腹水 6例,细菌性腹膜炎 1例.良、恶性腹水的诊断依据 : 恶性腹水有恶性肿瘤的原发灶,腹腔镜证实有腹膜转移或腹水找到恶性肿瘤细胞 ; 良性腹水有明确的病因,并经其它各项检查未发现恶性肿瘤征象.     

直肠癌及其癌旁组织DNA倍体的研究

大量研究表明 ,ＤＮＡ倍体的异常与肿瘤的发生发展及其临床病理特点密切相关[1] 。为了进一步探讨直肠癌的生物学行为 ,应用流式细胞仪 (ＦＣＭ )对 5 6例直肠癌新鲜组织及距癌灶下缘 3、4ｃｍ正常组织的ＤＮＡ指数 (ＤＩ)进行分析。1　材料与方法1.1　临床资料实验材料为 1999年 6月～ 2 0 0 1年 12月行Ｄｉｘｏｎ手术切除的 5 6例直肠癌标本 ,5 6例患者中男性 3 0例 ,女性 2 6例 ,年龄2 3～ 76岁 ,中位年龄 5 2岁。临床病理分期依据我国大肠癌协作组 1984年制定的标准。ＤｕｋｅｓＡ期 11例 ,ＤｕｋｅｓＢ期 2 3例 ,ＤｕｋｅｓＣ期 14例 ,…     

DNA倍体分析对乳腺癌恶性程度及预后评估的临床意义

目的探讨DNA倍体分析对乳腺癌恶性程度及预后评估的临床意义。方法用HPIAS-1000高清晰度彩色病理分析系统对有5年以上随访资料的120例乳腺癌患者进行组织学分级,高分化(Ⅰ级)48例,中等分化(Ⅱ级)44例,低分化(Ⅲ级)28例;并进行DNA倍体分析,即2倍体(2C)、3-4倍体(3-4C)、非整倍体(aneuploid,AN)。结果120例乳腺癌患者Ⅰ级与Ⅱ级、Ⅱ级与Ⅲ级、Ⅰ级与Ⅲ级之间,除3-4倍体外,2倍体及AN差异均有非常显著意义(P<0．01)。观察各级乳腺癌5年以上存活率与DNA倍体含量AN的出现率呈负相关,与2倍体的出现率呈正相关,与3-4倍体的出现率无相关意义。r=-0．9942,差异有非常显著意义(P<0．01)。结论DNA倍体分析对乳腺癌恶性程度及预后的评估有较高的临床意义。     

DNA倍体分析对乳腺癌恶性程度及预后评估的临床意义

目的 探讨DNA倍体分析对乳腺癌恶性程度及预后评估的临床意义。方法 用HPIAS-1000高清晰度彩色病理分析系统对有5年以上随访资料的120例乳腺癌患者进行组织学分级，高分化（Ⅰ级）48例，中等分化（Ⅱ级）44例，低分化（Ⅲ级）28例；并进行DNA倍体分析，即2倍体（2C）、3-4倍体（3-4C）、非整倍体（aneuploid，AN）。结果 120例乳腺癌患者Ⅰ级与Ⅱ级、Ⅱ级与Ⅲ级、Ⅰ级与Ⅲ级之间，除3-4倍体外，2倍体及AN差异均有非常显著意义（P〈0．01）。观察各级乳腺癌5年以上存活率与DNA倍体含量AN的出现率呈负相关，与2倍体的出现率呈正相关，与3-4倍体的出现率无相关意义。r=-0．9942，差异有非常显著意义（P〈0．01）。结论 DNA倍体分析对乳腺癌恶性程度及预后的评估有较高的临床意义。     

非何杰金氏淋巴瘤的恶性程度与DNA倍体关系的初步分析——36例流式细胞技术分析

 本文以石蜡包埋的淋巴结组织材料,采用流式细胞技术分析了36例组织学已确诊的非何杰金氏淋巴瘤的细胞倍体,并以5例慢性淋巴结炎为对照。按照NHL的Working Formulation的分类法:属低度恶性6例中的二倍体与非整倍体各3例。DNA指数均值为1.36±0.88。中度恶性11例中,二倍体5例,非整倍体6例,DNA指数均值为1.37±1.22。高度恶性19例,中二倍体1例,非整倍体18例,DNA指数均值1.80±1.02。经统计学处理:低度恶性比中度恶性P>0.05,无显著性差异,中度恶性与高度恶性P<0.05,有显著性差异。研究显示,通过流式细胞技术,分析NHL的倍体,能从分子生物学水平,克服光镜观察之不足处,比较客观的揭示NHL的恶性度,它有利于临床治疗及观察预后。     

肿瘤细胞DNA倍体分析中易于误解的若干问题

对肿瘤组织中肿瘤细胞ＤＮＡ倍体作分析 ,可为肿瘤病理及其临床诊疗和患者预后提供较客观的信息。然而 ,肿瘤细胞ＤＮＡ倍体分析中一些易于混淆和误解的问题 ,应引起注意。一、染色体倍体与ＤＮＡ倍体二倍体生物的体细胞 ,其染色体是由来自父方和来自母方的两个染色体组共同配对组成的 ,故称双倍体或二倍体 (ｄｉｐｌｏｉｄ ,简写 2ｎ)。染色体组(ｇｅｎｏｍｅ ,简写ｎ)是一个正常配子的全部染色体 ,若从基因角度讲 ,ｇｅｎｏｍｅ是指其全部染色体上遗传物质的总和 ,故ｇｅｎｏｍｅ又称基因组。单倍体 (ｍｏｎｏ ｐｌｏｉｄ ,简写 1ｎ)的细…     

食管癌DNA含量和细胞周期分析的临床意义

目的：研究食管癌和癌旁组织DNA含量、细胞周期分布与临床分期及组织病理学分级的关系和意义。方法：应用流式细胞术（FCM）测定40例食管癌、癌旁组织、10例正常食管粘膜细胞的DNA指数（DI）、S期细胞百分率（SPF）、异倍体率。结果：癌灶中心组织DI、SPF、异倍体率明显高于癌旁组织；DI、SPF、异倍体率明显高于无淋巴结转移。结论：DNA含量、异倍体率与癌中心灶密切相关，SPF可作为组织病理学分级、临床分期的一个参数。DI、SPF、异倍体率与患的临床信息结合，将有助于食管癌患的诊断、治疗及预后评价。     

Circulating Cell Free DNA and DNA Integrity Index as Discriminating Tools between Breast Cancer and Benign Breast Disease

Objectives: Early diagnosis of cancer remains a great challenge in the field of laboratory medicine. We investigated the ability of ccf DNA and DNA integrity index (DNA II) in differentiating benign from malignant breast diseases. Methods: Serum samples were collected from 50 patients with benign breast disease (BBD) and 50 newly diagnosed breast cancer (BC) patients, in addition to 50 control women. VEGF was measured by ELISA, while Real-time q-PCR was used to measure ccf DNA concentrations and to assess the concentrations of ALU repeats, both short fragments (115 bp) and long fragments (247 bp), then DNA II was calculated (all were done before and after radical mastectomy). Results: BC group showed significantly higher ccf DNA concentrations and DNA II compared to BBD and control groups, meanwhile, no statistically significant differences were found between BBD and control groups. Ccf DNA concentrations decreased significantly after surgery (P <0.001). Good AUC was found for ccf DNA (AUC=0.860), fair AUC was found for DNA II (AUC=0.727), while VEGF AUC failed to discriminate between BBD and BC cases. Conclusion: ccf DNA and DNA II could be used as excellent molecular biomarkers for early diagnosis of BC and for monitoring the efficiency of therapy in such patients. Utilizing these molecular markers would improve both the healthcare and economic burden of malignancy.     

Kaempferol Suppresses Proliferation and Induces Cell Cycle Arrest,Apoptosis, and DNA Damage in Breast Cancer Cells

Kaempferol is a flavonoid that has been extensively investigated owing to its antitumor effects. Nevertheless, little is known about its underlying mechanisms of action. We aimed to explore the role of kaempferol in breast cancer (BC), and thus we investigated how kaempferol suppresses the growth of BC cells. The cells were treated with kaempferol, and the effects on multiple cancer-associated pathways were evaluated. The MTS assay was used to study the cell growth inhibition induced by kaempferol. The cell cycle was analyzed by flow cytometry. Western blotting was used to analyze cellular apoptosis and DNA damage. We found that the proliferation of the triple-negative BC (TNBC) MDA-MB-231 cells was suppressed effectively by kaempferol. Interestingly, the suppressive effect of kaempferol on cell proliferation was stronger in MDA-MB-231 cells than in the estrogen receptor-positive BT474 cell line. Furthermore, after the treatment with kaempferol for 48 h, the population of cells in the G1 phase was significantly reduced, from 85.48% to 51.35%, and the population of cells in the G₂ phase increased markedly from 9.27% to 37.5%, which indicated that kaempferol contributed to the induction of G₂/M arrest. Kaempferol also induced apoptosis and DNA damage in MDA-MB-231 cells. Kaempferol increased the expression levels of H2AX, cleaved caspase 9, cleaved caspase 3, and p-ATM compared to those of the control group. Collectively, these results showed that kaempferol may be a potential drug for the effective treatment of TNBC.     

食管上皮癌变过程中DNA 含量及细胞周期调控因子表达的定量检测

 目的 探讨食管上皮癌变过程中DNA含量、细胞周期分布的变化及多个相关的细胞周期调控因子的表达状况及其意义. 方法 应用碘化丙啶染色和间接免疫荧光标记方法,采用流式细胞仪对30例食管癌组织及相应的癌旁组织进行定量检测. 结果 与正常黏膜和非典型增生组织相比,癌组织中DNA含量明显增高,异倍体细胞显著增加;G0/G1期细胞明显减少,而S期和G2/M期细胞显著增多,增殖指数（PI）高于癌旁组织;cyclinE、cdk2、p53、E2F基因蛋白过表达,两两之间皆有显著性正相关关系,抑癌基因p21^WAF1基因蛋白表达水平明显降低,与其余基因蛋白之间有明显的负相关. 结论 食管上皮癌变过程中,DNA含量及异倍体率增加,细胞周期分布发生了明显的改变,细胞周期相关基因蛋白表达异常.     

一种新的细胞周期分析方法

目的：采用DNA含量流式细胞术细胞周期分析方法,可分辨出G0/G1期、S期及G2/M期3个细胞周期群体,但无法区分G0/G1期、G2/M期细胞。方法：本研究建立的cyclin E A/DNA多参数流式细胞术。结果：可将细胞周期进行详细的分期,由传统的三分法（C0/G1期、S期以及G2/M期）为六分法（G0期、G1早期、G1晚期、S期、G2期以及M期）, 为细胞生物学研究,尤其是细胞增殖动力学提供新的,更为合理的分析方法。     

乳腺常见良,恶性病变的粘液组化观察

为了解乳腺癌与其它良性乳腺病变在分泌粘液性状上是否有诊断意义上的差异，本文运用ＰＡＳ、ＡＢ、ＨＩＤ３种粘液染色方法分别观察了乳腺的纤维腺瘤１１例，结构不良１１例，大导管乳头状瘤１０例，浸润性导管癌１２例及粘液癌１１例，共５５例进行粘液特性分析。结果表明良性病变３组唾液酸粘液的阳性率明显低于恶性病变２组；ＡＢ阳性率Ｐ＜０．０５，有显著性差异，提示乳腺肿瘤分泌唾液酸粘液增多与乳腺癌生长有关，且ＡＢ染色可做为乳腺癌诊断的辅助手段。     

乳腺癌细胞DNA含量分析

本文应用流式细胞术(Flow Cytomotry简称FCM)测定41例乳腺癌细胞DNA含量,分析结果:DNA指数(DI)范围为0.9～2.91,其中二倍体肿瘤13例,占31.7％,异倍体肿瘤28例,占68.3％。DI和病理组织学分级呈正相关系,随着肿瘤组织学分级的增高,DI也升高。但是,在目前研究中,DI尚不反映临床分期。DI和预后的关系,共观察19例,发现二倍体肿瘤的预后好,生存期长,异倍体肿瘤的预后差,生存期短。本文应用针吸肿瘤细胞和切除标本新鲜组织块检测DNA含量,发现两种取材方法所得数值无显著差别。因此,认为针吸细胞应用FCM的DNA检查可作为术前辅助诊断方法,为治疗提供参考。     

芒柄花素对不同亚型乳腺癌细胞增殖及细胞周期的影响

目的研究芒柄花素对人乳腺癌细胞MDA-MB-468、MCF-7、SK-BR-3的增殖及细胞周期影响。方法以三种乳腺癌细胞为研究对象,经MTT法观察不同浓度芒柄花素对乳腺癌细胞的增殖抑制情况;Western blot法检测增殖细胞核抗原（PCNA）表达情况;TUNEL法检测细胞凋亡情况;流式细胞仪观察细胞周期的改变情况。结果芒柄花素用药浓度大于20 μM时,抑制3种乳腺癌细胞的增殖, 降低PCNA蛋白的表达,G1期细胞比例均明显增多。结论芒柄花素可将细胞周期阻滞于G1期、减少PCNA蛋白的表达,从而抑制乳腺癌细胞增殖。     

Comparative DNA Analysis by Image Cytometry and Flow Cytometry in Non-small Cell Lung Cancer

To determine whether image cytometry (ICM) is advantageous for clinical DNA analyses of tumor cells, nuclear DNA contents measured by ICM were compared with those by flow cytometry (FCM), using 46 samples of non-small cell lung cancers. ICM was performed on smear specimens of fresh materials (f-ICM) and cell suspensions obtained from paraffin-embedded tumors (p-ICM). The same cell suspensions were also analyzed by FCM (p-FCM). Aneuploid rates/coefficient of variation (CV) of f-ICM, p-ICM, and p-FCM were 76.1/4.90, 71.7/5.01 and 60.9/5.31%, respectively. There was a high correlation in the DNA indices between p-ICM and p-FCM (r=0.80). In the comparative DNA analysis, there were seven discordant samples. Six of them were estimated as aneuploid by p-ICM, but they were miscounted as diploid or undefinablc (impossible) by p-FCM. This was caused by measuring condensed nuclei or debris. All "impossible" samples in p-FCM were squamous cell carcinoma with necrosis. In cell cycle analysis, the S and S+G2/M phase fractions in diploid samples were higher in p-ICM than those in p-FCM ( P < 0.005), because the GO/G1 phase (2N) fraction presented by FCM was composed of cancer and non-malignant cells in diploid cancers. In ICM, they can be separately measured by means of morphological selection. These findings indicated that ICM is superior to FCM, especially for the practical DNA measurement of a few cancer cells and in the evaluation of the proliferation rates.     

前列腺上皮内瘤DNA 倍体、细胞周期的分析

 目的 探索DNA倍体类型、细胞周期在前列腺上皮内瘤中诊断和鉴别诊断的价值。方法 应用细胞图像分析技术检测前列腺上皮内瘤（PIN）细胞的平均DNA含量、DNA平均倍体值及细胞周期的各个时相的变化。同时与良性前列腺增生症（BPH）、前列腺癌（PCa）比较。结果 PIN组异倍体率为70％，前列腺上皮内瘤DNA平均倍体值、G0／G1期细胞比率、S期细胞比率介于良性前列腺增生症和前列腺癌之间。结论 DNA异倍体、较高S期细胞比率是前列腺上皮内瘤重要生物学特性。对于前列腺上皮内瘤的鉴别诊断具有一定参考价值。