目的:探讨Annexin V FITC/PI流式细胞术在检测姜黄素诱导的肝星状细胞(HSC)凋亡中的应用。方法:不同浓度姜黄素作用肝星状细胞株HSC-T6 24h,用Annexin V FITC/PI流式细胞术检测HSC凋亡。结果:姜黄素浓度为10μmol/L时,早期凋亡细胞开始...目的:探讨Annexin V FITC/PI流式细胞术在检测姜黄素诱导的肝星状细胞(HSC)凋亡中的应用。方法:不同浓度姜黄素作用肝星状细胞株HSC-T6 24h,用Annexin V FITC/PI流式细胞术检测HSC凋亡。结果:姜黄素浓度为10μmol/L时,早期凋亡细胞开始增多,至30μmol/L时,坏死细胞开始增多,随姜黄素浓度增高,各组凋亡细胞随之增多。0、10、20、30、40μmol/L5组的凋亡率分别是(3.81±0.64)%、(6.52±1.88)%、(11.61±2.79)%、(52.03±4.38)%、(87.11±12.62)%,与0μmol/L组比较,20、30、40μmol/L3组的凋亡率差异有统计学意义(P<0.05)。结论:Annexin V FITC/PI流式细胞术可定量检测姜黄素诱导的HSC早期凋亡。展开更多
AIM: To achieve a better understanding of the pathogenesis of new type gosling viral enteritis virus (NGVEV) and the relationship between NGVEV and host cells. METHODS: The apoptosis of duck embryo fibroblasts (DEF) i...AIM: To achieve a better understanding of the pathogenesis of new type gosling viral enteritis virus (NGVEV) and the relationship between NGVEV and host cells. METHODS: The apoptosis of duck embryo fibroblasts (DEF) induced by NGVEV was investigated by fluorescence-activated cell sorter (FACS) and fluorescence microscope after the cells were stained with Annexin V-FITC and propidium iodide (PI). RESULTS: By staining cells with a combination of fluorescein annexin V-FITC and PI, it is possible to distinguish and quantitatively analyze non-apoptotic cells (Annexin V-FITC negative/PI negative), early apoptotic cells (Annexin V-FITC positive/PI negative), late apoptotic/necrotic cells (Annexin V-FITC positive/ PI positive) and dead cells (Annexin V-FITC negative/PI positive) through flow cytometry and fluorescence microscope. The percentage of apoptotic cells increased with the incubation time and reached a maximum at 120 h after infection, while the percentage of non- apoptotic cells decreased.展开更多
Objective To study the findings of serum antibodies against annexin V, prothrombin, ph-inositol, ph-acid, ph-ethanolamine, ph-serine, ph-glycerol, cardiolipin, and beta2-glycoprotein I and analyze the trophoblast anne...Objective To study the findings of serum antibodies against annexin V, prothrombin, ph-inositol, ph-acid, ph-ethanolamine, ph-serine, ph-glycerol, cardiolipin, and beta2-glycoprotein I and analyze the trophoblast annexin V receptors Methods Sera from 156patients aged 26-41 years with recurrent pregnancy loss （3-7 times） were investigated. Eighty-four fertile healthy women aged 24-38 years were included in a control group. ELISA methods were used for detecting a panel of sera anti-phospholipid antibodies. Immunolocalization of annexin V receptors in 143 trophoblast specimens of 156 patients was investigated by the immunofluorescence technique using Annexin V-FITC, Apoptosis and Annexin V-CY3 commercial kits. Results Positivity for anti-phospholipid antibodies mainly against ph-serine, ph- ethanolamine, and ph-inositol was found together in 80. 8%（126 out of 156 patients）, anti-prothrombin antibodies in 12% （18）, and anti-annexin V antibodies in 13. 5% （21） women. No significant levels of anti-phospholipid antibodies were found in 6 controls. Placenta immunohistopathology also exhibited some changes manifested by the presence of apoptotic and necrotic cells in trophoblast, and very few microthrombotization in some intervillous spaces. Conclusion Our detailed study demonstrated the prevalence of majority of antiphospholipid antibodies as a high risk factor for repeated reproductive failure. Very low microthrombosis in placentas could be explained by the changes of haemocoagulation properties out of uterus.展开更多
目的制备人膜联蛋白 V(Annexin V),应用联肼尼克酰胺(HYNIC)偶联后进行^(99)Tc^m 标记,评价^(99)Tc^m-HYNIC-Annexin V 在健康小鼠体内的分布。方法采用基因工程方法构建 Annexin V 的原核载体并诱导其表达,与自行合成的双功能螯合剂 HY...目的制备人膜联蛋白 V(Annexin V),应用联肼尼克酰胺(HYNIC)偶联后进行^(99)Tc^m 标记,评价^(99)Tc^m-HYNIC-Annexin V 在健康小鼠体内的分布。方法采用基因工程方法构建 Annexin V 的原核载体并诱导其表达,与自行合成的双功能螯合剂 HYNIC 偶联,并进行^(99)Tc^m 标记。测定^(99)Tc^m-HYNIC-Annexin V 的标记率、放化纯,并研究其在健康小鼠体内的生物分布特性。结果 Annexin V在大肠杆菌中获得稳定表达,经十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)及 Western Blot-ting 检测,得到纯度较高的蛋白质。Annexin V 经 HYNIC 偶联后,其^(99)Tc^m 标记率可达90%左右,放化纯>90%。小鼠体内分布结果表明,^(99)Tc^m-HYNIC-Annexin V 血液清除迅速,主要从肾脏和肝脏排泄。结论 Annexin V 可在原核生物中稳定表达。其与 HYNIC 偶联后,^(99)Tc^m 标记率和放化纯较高,方法简单,条件温和。展开更多
目的研究^(99)Tc^m-联肼尼克酰胺-膜联蛋白 V(HYNIC-Annexin V)的制备及其在荷瘤小鼠体内生物分布特性和活体显像。方法用双功能螯合剂法,以^(99)Tc^m 标记 Annexin V,经高效液相色谱仪分离纯化并检测产物的标记率、放化纯及稳定性。建...目的研究^(99)Tc^m-联肼尼克酰胺-膜联蛋白 V(HYNIC-Annexin V)的制备及其在荷瘤小鼠体内生物分布特性和活体显像。方法用双功能螯合剂法,以^(99)Tc^m 标记 Annexin V,经高效液相色谱仪分离纯化并检测产物的标记率、放化纯及稳定性。建立荷 S180肉瘤小鼠模型,于20～25g/只昆明种小白鼠右前腋皮下接种 S180肉瘤细胞1周。荷瘤小鼠在环磷酰胺腹膜内化疗72 h 后,尾静脉注射7.4 MBq(200μl)^(99)Tc^m-HYNIC-Annexin V,分别于5、30 min 和1、3、6 h 进行显像和体内生物分布测定。实验结果应用 SPSS 12.0统计软件进行分析。结果 ^(99)Tc^m-HYNIC-Annexin V 的标记率达95%,放化纯为99%。荷瘤小鼠注射^(99)Tc^m-HYNIC-Annexin V 后6 h 显像,可见肿瘤组织放射性浓聚明显异常。体内生物分布实验示,注射显像剂后6 h 肿瘤组织的放射性摄取最高[每克组织百分注射剂量率(%ID/g)为1.59±0.44],与其他时相比较,差异有显著性(P<0.05)。^(99)Tc^m-HYNIC-Annexin V 主要浓聚于肾、肺和肝,经肾脏排泄;其在血液中清除速度快,注射后30 min,血液放射性摄取[(1.59±0.50)%ID/g]较注射后5 min 时[(8.85±2.65)%ID/g]减少80%(P<0.05)。注射显像剂后6 h,肿瘤/肌肉放射性摄取比值(3.73±1.42)高于肿瘤/血液(2.80±0.54)。结论 ^(99)Tc^m-HYNIC-Annexin V 活体细胞凋亡显像可应用于荷瘤小鼠模型,其临床应用有待进一步研究。展开更多
目的评价人膜联蛋白 V(H-Annexin V)对血栓的亲和性,对^(99)Tc^m直接标记 H-Annexin V与通过联肼尼克酰胺(HYNIC)螯合标记 Annexin V 进行比较。方法在3种不同的反应条件下,分别用^(99)Tc^mO_4^-直接标记 Annexin V、H-Annexin V 及 HYN...目的评价人膜联蛋白 V(H-Annexin V)对血栓的亲和性,对^(99)Tc^m直接标记 H-Annexin V与通过联肼尼克酰胺(HYNIC)螯合标记 Annexin V 进行比较。方法在3种不同的反应条件下,分别用^(99)Tc^mO_4^-直接标记 Annexin V、H-Annexin V 及 HYNIC-Annexin V,然后测定标记率、稳定性及对兔股动脉血栓模型进行γ显像,比较损伤侧(T,血栓)及对侧(NT)的放射性比值。结果在还原剂足量的情况下,^(99)Tc^mO_4^-直接标记 H-Annexin V 方法简便,反应条件易于控制,^(99)Tc^m-H-Annexin V标记率达95%以上,17 h 后测放化纯仍>80%,血栓显像示动脉损伤处有明显的放射性浓聚,损伤处平均放射性计数为健侧放射性计数的3.714倍。结论 ^(99)Tc^m-H-Annexin V 与血栓具有很好的亲和力,标记率及稳定性高,标记方法较为简单。展开更多
文摘目的:探讨Annexin V FITC/PI流式细胞术在检测姜黄素诱导的肝星状细胞(HSC)凋亡中的应用。方法:不同浓度姜黄素作用肝星状细胞株HSC-T6 24h,用Annexin V FITC/PI流式细胞术检测HSC凋亡。结果:姜黄素浓度为10μmol/L时,早期凋亡细胞开始增多,至30μmol/L时,坏死细胞开始增多,随姜黄素浓度增高,各组凋亡细胞随之增多。0、10、20、30、40μmol/L5组的凋亡率分别是(3.81±0.64)%、(6.52±1.88)%、(11.61±2.79)%、(52.03±4.38)%、(87.11±12.62)%,与0μmol/L组比较,20、30、40μmol/L3组的凋亡率差异有统计学意义(P<0.05)。结论:Annexin V FITC/PI流式细胞术可定量检测姜黄素诱导的HSC早期凋亡。
基金The National Natural Science Foundation of China, No. 39970561The Key Projects in the National Science and Technology Pillar Program, 2007Z06-017+2 种基金Program for New Century Excellent Talents from Universities, Chinese Ministry of Education, No. NCET-04-0906/NCET-06-0818Fund of the Discipline Leaders of Sichuan Province, No. SZD0418Culture Fund for Excellent Doctoral Dissertations of Sichuan Agricultural University, 2008scybpy-1
文摘AIM: To achieve a better understanding of the pathogenesis of new type gosling viral enteritis virus (NGVEV) and the relationship between NGVEV and host cells. METHODS: The apoptosis of duck embryo fibroblasts (DEF) induced by NGVEV was investigated by fluorescence-activated cell sorter (FACS) and fluorescence microscope after the cells were stained with Annexin V-FITC and propidium iodide (PI). RESULTS: By staining cells with a combination of fluorescein annexin V-FITC and PI, it is possible to distinguish and quantitatively analyze non-apoptotic cells (Annexin V-FITC negative/PI negative), early apoptotic cells (Annexin V-FITC positive/PI negative), late apoptotic/necrotic cells (Annexin V-FITC positive/ PI positive) and dead cells (Annexin V-FITC negative/PI positive) through flow cytometry and fluorescence microscope. The percentage of apoptotic cells increased with the incubation time and reached a maximum at 120 h after infection, while the percentage of non- apoptotic cells decreased.
基金This study was supported by grants of Czech Ministry of Health (VU 96/69).
文摘Objective To study the findings of serum antibodies against annexin V, prothrombin, ph-inositol, ph-acid, ph-ethanolamine, ph-serine, ph-glycerol, cardiolipin, and beta2-glycoprotein I and analyze the trophoblast annexin V receptors Methods Sera from 156patients aged 26-41 years with recurrent pregnancy loss （3-7 times） were investigated. Eighty-four fertile healthy women aged 24-38 years were included in a control group. ELISA methods were used for detecting a panel of sera anti-phospholipid antibodies. Immunolocalization of annexin V receptors in 143 trophoblast specimens of 156 patients was investigated by the immunofluorescence technique using Annexin V-FITC, Apoptosis and Annexin V-CY3 commercial kits. Results Positivity for anti-phospholipid antibodies mainly against ph-serine, ph- ethanolamine, and ph-inositol was found together in 80. 8%（126 out of 156 patients）, anti-prothrombin antibodies in 12% （18）, and anti-annexin V antibodies in 13. 5% （21） women. No significant levels of anti-phospholipid antibodies were found in 6 controls. Placenta immunohistopathology also exhibited some changes manifested by the presence of apoptotic and necrotic cells in trophoblast, and very few microthrombotization in some intervillous spaces. Conclusion Our detailed study demonstrated the prevalence of majority of antiphospholipid antibodies as a high risk factor for repeated reproductive failure. Very low microthrombosis in placentas could be explained by the changes of haemocoagulation properties out of uterus.
文摘目的制备人膜联蛋白 V(Annexin V),应用联肼尼克酰胺(HYNIC)偶联后进行^(99)Tc^m 标记,评价^(99)Tc^m-HYNIC-Annexin V 在健康小鼠体内的分布。方法采用基因工程方法构建 Annexin V 的原核载体并诱导其表达,与自行合成的双功能螯合剂 HYNIC 偶联,并进行^(99)Tc^m 标记。测定^(99)Tc^m-HYNIC-Annexin V 的标记率、放化纯,并研究其在健康小鼠体内的生物分布特性。结果 Annexin V在大肠杆菌中获得稳定表达,经十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)及 Western Blot-ting 检测,得到纯度较高的蛋白质。Annexin V 经 HYNIC 偶联后,其^(99)Tc^m 标记率可达90%左右,放化纯>90%。小鼠体内分布结果表明,^(99)Tc^m-HYNIC-Annexin V 血液清除迅速,主要从肾脏和肝脏排泄。结论 Annexin V 可在原核生物中稳定表达。其与 HYNIC 偶联后,^(99)Tc^m 标记率和放化纯较高,方法简单,条件温和。
文摘目的研究^(99)Tc^m-联肼尼克酰胺-膜联蛋白 V(HYNIC-Annexin V)的制备及其在荷瘤小鼠体内生物分布特性和活体显像。方法用双功能螯合剂法,以^(99)Tc^m 标记 Annexin V,经高效液相色谱仪分离纯化并检测产物的标记率、放化纯及稳定性。建立荷 S180肉瘤小鼠模型,于20～25g/只昆明种小白鼠右前腋皮下接种 S180肉瘤细胞1周。荷瘤小鼠在环磷酰胺腹膜内化疗72 h 后,尾静脉注射7.4 MBq(200μl)^(99)Tc^m-HYNIC-Annexin V,分别于5、30 min 和1、3、6 h 进行显像和体内生物分布测定。实验结果应用 SPSS 12.0统计软件进行分析。结果 ^(99)Tc^m-HYNIC-Annexin V 的标记率达95%,放化纯为99%。荷瘤小鼠注射^(99)Tc^m-HYNIC-Annexin V 后6 h 显像,可见肿瘤组织放射性浓聚明显异常。体内生物分布实验示,注射显像剂后6 h 肿瘤组织的放射性摄取最高[每克组织百分注射剂量率(%ID/g)为1.59±0.44],与其他时相比较,差异有显著性(P<0.05)。^(99)Tc^m-HYNIC-Annexin V 主要浓聚于肾、肺和肝,经肾脏排泄;其在血液中清除速度快,注射后30 min,血液放射性摄取[(1.59±0.50)%ID/g]较注射后5 min 时[(8.85±2.65)%ID/g]减少80%(P<0.05)。注射显像剂后6 h,肿瘤/肌肉放射性摄取比值(3.73±1.42)高于肿瘤/血液(2.80±0.54)。结论 ^(99)Tc^m-HYNIC-Annexin V 活体细胞凋亡显像可应用于荷瘤小鼠模型,其临床应用有待进一步研究。
文摘目的评价人膜联蛋白 V(H-Annexin V)对血栓的亲和性,对^(99)Tc^m直接标记 H-Annexin V与通过联肼尼克酰胺(HYNIC)螯合标记 Annexin V 进行比较。方法在3种不同的反应条件下,分别用^(99)Tc^mO_4^-直接标记 Annexin V、H-Annexin V 及 HYNIC-Annexin V,然后测定标记率、稳定性及对兔股动脉血栓模型进行γ显像,比较损伤侧(T,血栓)及对侧(NT)的放射性比值。结果在还原剂足量的情况下,^(99)Tc^mO_4^-直接标记 H-Annexin V 方法简便,反应条件易于控制,^(99)Tc^m-H-Annexin V标记率达95%以上,17 h 后测放化纯仍>80%,血栓显像示动脉损伤处有明显的放射性浓聚,损伤处平均放射性计数为健侧放射性计数的3.714倍。结论 ^(99)Tc^m-H-Annexin V 与血栓具有很好的亲和力,标记率及稳定性高,标记方法较为简单。
