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| 1 | Sensitivity and specificity of different staining methods to monitor apoptosis induced by oxidative stress in adherent cells显示文摘Objective To study the sensitivity and specificity of different staining methods to monitor apoptosis induced by oxidative stress in adherent cells.Methods Sensitivity and specificity of several common methods for apoptosis determination were evaluated (Apo2.7-expression, Annexin V-binding, TUNEL-reaction, poly-(ADP-ribose)-polymerase-(PARP) cleavage and single-stranded-DNA (ssDNA) staining). Apoptosis was induced by oxidative stress generated by hydrogen peroxide in 3 cultured cells types growing as adherent monolayer (MiaPaCa-2, Hep-G2 and human skin fibroblasts), necrosis was induced by depletion of cellular ATP using sodium azide. Cells positively stained by the respective apoptosis assay were quantified and alterations of cell morphology were monitored by fluorescence microscopy. The date was analyzed by one-way analysis of variance and significance test of correlation coefficient.Results One hour after apoptosis induction significant cell fractions were positively stained for ssDNA (33% with MiaPaCa-2 cells, 35% with Hep-G2 cells, 56% with human skin fibroblasts). PARP-cleavage was less sensitive compared to the ssDNA-staining. Apo2.7-expression, Annexin V-binding and TUNEL-reaction were not applicable to detect early apoptosis induced by oxidative stress (below 2 hours), but were efficiently monitoring late apoptosis. Specificity of ssDNA-staining was complete with each cell type even 4 hs after induction of necrosis by the highest sodium azide concentration. In contrast, the same experimental conditions resulted in 50%-90% positively stained necrotic cells by using Apo2.7-expression, TUNEL-reaction or AnnexinV-binding. Surprisingly, specificity of PARP-cleavage was highly depending on the respective cell type.Conclusions Our study prove that among the five methods investigated only ssDNA-staining allowed to completely differentiate apoptosis from necrosis, and is thus suitable to reliably detect early as well as late apoptosis. Therefore, the ssDNA-staining may be used as reference method to clearly identify apoptosis induced by oxidative stress in adherent cells. The TUNEL-reaction, annexin-V-binding and Apo-2.7-expression may be used to quantify the number of apoptotic and necrotic cells especially at later stages but without discrimination of apoptosis and primary or secondary necrosis. | 赵进顺 Alexandra Schmid-Kotsas Hans-Juergen Gross Adolf Gruenert Max G. Bachem | 2003 | Chinese Medical Journal2003，，12： | 4 |
| 2 | Identification,culture,and characterization of pancreatic stellate cells in rats and humans显示文摘 | Bachem MG Schneider E Gross H | | Gastroenterology0，，： | 2 |
| 3 | 低频低强度超声波诱导白血病细胞株K562凋亡的研究显示文摘目的:研究低频低强度超声波诱导白血病细胞株K562凋亡的作用及其机制。方法:取对数生长期的K562细胞,分成实验组和对照组,用MTT法计算细胞存活率,流式细胞仪判断细胞凋亡,瑞氏染色和透射电镜观察细胞照射后有无出现凋亡,分光光度法检测照射前后K562细胞Caspase-3的活性改变。结果:超声照射各组间细胞存活率与对照组相比有统计学差异,但组间无差异;瑞氏染色光镜下可见0.03 W组和0.10W组细胞出现细胞核固缩等早期凋亡的改变,0.25W组出现凋亡小体等典型凋亡改变;流式细胞仪检测0.25 W组凋亡率为38.87%;照射前后K562细胞Caspase-3的活性无差异。结论:低频低强度超声波对白血病细胞株K562有诱导凋亡作用,与照射强度和时间无明显相关。低频低强度超声波诱导K562的凋亡可能不通过Caspase-3的途径。 | 马燕 陈宝安 吴巍 姜藻 钱梦騄 Bachem M G 周韶霞 赵进顺 | 2006 | 东南大学学报(医学版)2006，25，3： | 2 |
| 4 | Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP: Analysis of gene expression during potato tuber development显示文摘 | Bachem C W B Van der Hoeven R S de Bruijn S M | 1996 | Plant J1996，9，： | 1 |
| 5 | Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP:Analysis of gene expression during potato tuber development显示文摘 | Bachem C W B van der Hoeven R S de Bruijn S M Vreugdenhil D Zabeau M Visser R G F | 1996 | Plant J1996，9，： | 1 |
| 6 | Pancreat- ic carcinoma cells induce fibrosis by stimulating proliferation and matrix synthesis of stellate cells 显示文摘 | Bachem MG Schiinemann M Ramadani M | 2005 | Gastroenterology2005，128，： | 1 |
| 7 | Isolation and characterization of a novel potato AuxirdIndole-3-Acetic Acid family member(StIAA2) that is involved in petiole hy ponasty and shoot morphogenesis 显示文摘 | KLOOSTERMAN B VISSER R G BACHEM C W | 2006 | Plant Physiol Biochem2006，44，1112： | 1 |
| 8 | Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP:Analysis of gene expression during potato tuber development显示文摘 | Bachem C W B van der Heoven R S de Bruijn S M | 1996 | Plant Journal1996，9，5： | 1 |
| 9 | A physical, yet simple, small-signal equivalent circuit for the heterojunction bipolar transistor 显示文摘 | GOBERT Y TASKER P J BACHEM K H | 1997 | IEEE Trans Microwave Theory and Techniques1997，45，1： | 1 |
| 10 | Transcript imagingwith cDNA-AFLP:a step-by-step protocol 显示文摘 | Bachem CWB Oomen RJF Visser RGF | 1998 | Plant Molecu-lar Biology Report1998，16，： | 1 |
| 11 | A physical, yet simple, small-signal equivalent circuit for the hetero- juction bipolar transistor 显示文摘 | Gobert Y Tasker P J Bachem K H | 1997 | IEEE Transactions on Microwave Theory and Techniques1997，45，1： | 1 |
| 12 | Functional genomic analysis of potato tuber life-cycle显示文摘 | Bachem CWB van der Hoeven RS Lucker J | 2000 | Potato Res2000，43，： | 1 |
| 13 | Visualization of Differential Gene Expression Using a Novel Method of RNA Fingerprinting Based on AFLP: Analysis of Gene Expression During Potato Tuber Development显示文摘 | BACHEM G W B | 1996 | Plant Journal1996，9，5： | 1 |
| 14 | Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP:analysis of gene expression during potato tuber development显示文摘 | BACHEM C W B VAN DER HOEVEN R S DE BRUIJN S M VREUGDENHIL D ZABEAU M VISSER R G F | 1996 | Plant Journal1996，9，5： | 1 |
| 15 | Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP: Analysis of gene expression during potato tuber development显示文摘 | Bachem CWB Hoeven RSVD Bruijn SMD | 1996 | The Plant Journal1996，9，5： | 1 |
| 16 | Self-compensation in Mg doped p-type GaN grown by MOCVD显示文摘 | Obloh H Bachem K H Kaufmann U | | 0，，01： | 1 |
| 17 | Molecular mechanisms of fibrogenesis-A homage to the role of activatefat-storing cells显示文摘 | Grossncr AM Bachem MG | 1995 | Digestion1995，56，1： | 1 |
| 18 | Vi-sualization of differential gene Expression using a novel method ofRNA fingerprinting based on AFLP:analysis of gene expressionduring potato tuber development显示文摘 | Bachem C W B Van der Hoeven R S de Bruijn S M | 1996 | The Plant Journal1996，9，5： | 1 |
| 19 | Identification,culture,and characterization of pancreatic stellate cells in rat and humans显示文摘 | Bachem MG Schneider E Gross H | 1998 | Gastroenterology1998，115，2： | 1 |
| 20 | Identification,culture,and characterization of pancreatic stellate cells in rats and humans显示文摘 | Bachem MG Schneider E Gross H | 1998 | Gastroenterology1998，115，2： | 1 |
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