Globozoospermia is a severe form of teratozoospermia characterized by round-headed spermatozoa with an absent acrosome, an aberrant nuclear membrane and midpiece defects. Globozoospermia is diagnosed by the presence o...Globozoospermia is a severe form of teratozoospermia characterized by round-headed spermatozoa with an absent acrosome, an aberrant nuclear membrane and midpiece defects. Globozoospermia is diagnosed by the presence of 100% round-headed spermatozoa on semen analysis, and patients with this condition are absolutely infertile. The objective of this study was to investigate the differences in protein expression between human round- headed and normal spermatozoa. Two-dimensional （2-D） fluorescence difference gel electrophoresis （DIGE） coupled with mass spectrometry （MS） was used in this study. Over 61 protein spots were analysed in each paired normal/round-headed comparison, using DIGE technology along with an internal standard. In total, 35 protein spots identified by tandem mass spectrometry （MS/MS） exhibited significant changes （paired t-test, P 〈 0.05） in the expression level between normal and round-headed spermatozoa. A total of nine proteins were found to be upregulated and 26 proteins were found to be downregulated in round-headed spermatozoa compared with normal spermatozoa. The differentially expressed proteins that we identified may have important roles in a variety of cellular processes and structures, including spermatogenesis, cell skeleton, metabolism and spermatozoa motility.展开更多
Objective To identify the sperm membrane proteins that are associated with antisperm antibody Methods Using antisperm antibody positive serum through unidimensional polyacrylamide gel electrophoresis and 2-dimensi...Objective To identify the sperm membrane proteins that are associated with antisperm antibody Methods Using antisperm antibody positive serum through unidimensional polyacrylamide gel electrophoresis and 2-dimensional gel electrophoresis followed by Western blot analysis to determine the molecular weights (MW) and isoelectric points (pI) of sperm membrane proteins that are associated with antisperm antibody. Results Eight kinds of MW with more than ten sperm membrane proteins can be recognized by antisperm antibody positive serum, of which the MWs and pI were 23 kD, 31 kD, 32 kD, 34 kD, 41 kD, 51 kD, 60 kD, 78 kD and 5.3, 5.5,5.7, 5.0, 5.3, 5.8, 6.0, 5.5～6.2, 4.6,5.1,5.5～5.8 respectively. The identification ratios of the sperm membrane proteins on 78 kD (60.7%), 60 kD (71.4%), 51 kD (14.9%) and 23 kD (14.29%) were higher. Conclusion The sperm membrane proteins with MW of 78 kD, 60 kD, 51 kD and 23 kD were associated with antisperm antibody and immunological infertility. Two- dimensional gel electrophoresis and Western blotting can precisely identify the sperm membrane proteins that are associated with antisperm antibody.展开更多
目的:筛选人急性髓系白血病细胞株HL-60甲基化沉默基因,为揭示白血病的发病机制及防治提供科学依据。方法:应用荧光差异双向凝胶电泳(two-dimensional fluorescence difference gel electro-phoresis,F-2D-DIGE)分离甲基转移酶抑制剂5-...目的:筛选人急性髓系白血病细胞株HL-60甲基化沉默基因,为揭示白血病的发病机制及防治提供科学依据。方法:应用荧光差异双向凝胶电泳(two-dimensional fluorescence difference gel electro-phoresis,F-2D-DIGE)分离甲基转移酶抑制剂5-杂氮-2-脱氧胞苷(5-aza-2-dC)处理与未处理的HL-60细胞的总蛋白质,Decyder和PDquest图像分析软件识别差异表达蛋白质点,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)技术鉴定差异表达蛋白质。结果:建立了5-aza-2-dC处理与未处理的HL-60细胞蛋白质的F-2D-DIGE图谱,图像分析识别了53个差异表达的蛋白质点,质谱分析鉴定了35个差异表达的蛋白质,其中32个蛋白质在5-aza-2-dC处理后的HL-60细胞中表达上调,3个蛋白质表达下调。结论:35个差异表达蛋白可能与甲基化相关,为白血病的表观遗传学研究提供了有价值的信息。展开更多
AIM: To establish and compare serum proteomic of diabetic retinopathy (DR) patients in various phases and discuss pathogenesis of DR so as to find out possible serum specific molecular markers for early diagnosis of D...AIM: To establish and compare serum proteomic of diabetic retinopathy (DR) patients in various phases and discuss pathogenesis of DR so as to find out possible serum specific molecular markers for early diagnosis of DR. METHODS: Thirty-two subjects were divided into four groups: one group of eight type 2 diabetes mellitus (T2DM) patients without apparent DR (No-DR, NDR), one group of eight T2DM patients with non-proliferative diabetic retinopathy (NPDR), one group of eight T2DM patients with proliferative diabetic retinopathy(PDR) and one group of eight healthy volunteer participants. Two dimensional fluorescence difference gel electrophoresis (2D-DIGE) was applied to establish differential protein expression profiles in four groups. Matrix-assisted laser desorption/ionization time of flight tandem mass spectrometry (MALDI-TOF-TOF MS) was applied to identify mass spectrometry of differential proteins and analyze follow-up bioinformatics. RESULTS: 2D-DIGE maps of serum protein were satisfactory obtained from NDR, NPDR, PDR and normal control groups. Twenty-six different proteins spots were screened(the volume ratio was > 1.5 based on DeCyder software analysis). Twenty-four of them were verified and two of them were not. Fifteen proteins were verified. Most of them were high-abundant proteins in serum. The four relatively low-abundant ones were beta 2-glycoprotein I (beta (2)-GPI), alpha2-HS-glycoprotein (AHSG), alpha1-acid glycoprotein (alpha(1)-AGP) and apolipoprotein A-1 (apo A-1). beta (2)-GPI expression was gradually increased in the development of DR but unrelated to the severity of DR. The volume ratio of beta (2)-GPI is 1.54, 2.43, and 2.84 in NDR, NPDR and PDR group respectively compared with normal control group. CONCLUSION: Serum proteomic analysis of 2D-DIGE combined with MALDI-TOF-TOF MS is feasible to be applied in the study of DR. 13 2-GPI probably takes part in the process of DR occurrence and development and it could be a candidate biomarker on DR diagnosis in early phase.展开更多
The prefrontal cortex is involved in the regulation and control of substance addiction-related cognitive, behavioral, and emotional changes. The present study identified prefrontal cortex protein profiles in morphine-...The prefrontal cortex is involved in the regulation and control of substance addiction-related cognitive, behavioral, and emotional changes. The present study identified prefrontal cortex protein profiles in morphine-addicted rats; these were subsequently compared with normal rats. Results showed 87 protein spots with differentially expressed levels in the morphine addiction group, with the majority located in meta acid zones at pH 4.2-6.8 and having a molecular weight of 30-110 kDa In addition, 2 protein spots were identified as being associated with neurotoxicity （Snap25 isoform β-Snap25 of synaptosomal-associated protein 25 and βactin）.展开更多
目的:探讨双向差异凝胶电泳(two dimension difference gel electrophoresis,2D-DIGE)的实验方法,分析比较其优缺点,概述其在微生物研究中的应用。方法:以两株不同血清型的副溶血性弧菌为样本,通过2D-DIGE实验,得到两组蛋白质荧光染色...目的:探讨双向差异凝胶电泳(two dimension difference gel electrophoresis,2D-DIGE)的实验方法,分析比较其优缺点,概述其在微生物研究中的应用。方法:以两株不同血清型的副溶血性弧菌为样本,通过2D-DIGE实验,得到两组蛋白质荧光染色分布图,与银染的效果比较。结果:直观的看到两株细菌的蛋白质差异,且可分析样品丰度变化。结论:该方法有一定的局限性,但却是研究蛋白质组丰度变化的强有力工具。展开更多
基金Acknowledgment We thank Beijing Proteome Research Center, （Beijing, China） for its enthusiastic technological support and for the theory of 2-D DIGE. We also thank（Changsha, China） College of Life Sciences at Hunan Normal University for supporting the MS technology. Finally, we are very grateful to our collaborators for their help, as well as their valuable discussions and suggestions during the course of this work. This work was supported by two grants from the National Natural Science Foundation of China （NO. 30170480 and NO. 30470884）.
文摘Globozoospermia is a severe form of teratozoospermia characterized by round-headed spermatozoa with an absent acrosome, an aberrant nuclear membrane and midpiece defects. Globozoospermia is diagnosed by the presence of 100% round-headed spermatozoa on semen analysis, and patients with this condition are absolutely infertile. The objective of this study was to investigate the differences in protein expression between human round- headed and normal spermatozoa. Two-dimensional （2-D） fluorescence difference gel electrophoresis （DIGE） coupled with mass spectrometry （MS） was used in this study. Over 61 protein spots were analysed in each paired normal/round-headed comparison, using DIGE technology along with an internal standard. In total, 35 protein spots identified by tandem mass spectrometry （MS/MS） exhibited significant changes （paired t-test, P 〈 0.05） in the expression level between normal and round-headed spermatozoa. A total of nine proteins were found to be upregulated and 26 proteins were found to be downregulated in round-headed spermatozoa compared with normal spermatozoa. The differentially expressed proteins that we identified may have important roles in a variety of cellular processes and structures, including spermatogenesis, cell skeleton, metabolism and spermatozoa motility.
文摘Objective To identify the sperm membrane proteins that are associated with antisperm antibody Methods Using antisperm antibody positive serum through unidimensional polyacrylamide gel electrophoresis and 2-dimensional gel electrophoresis followed by Western blot analysis to determine the molecular weights (MW) and isoelectric points (pI) of sperm membrane proteins that are associated with antisperm antibody. Results Eight kinds of MW with more than ten sperm membrane proteins can be recognized by antisperm antibody positive serum, of which the MWs and pI were 23 kD, 31 kD, 32 kD, 34 kD, 41 kD, 51 kD, 60 kD, 78 kD and 5.3, 5.5,5.7, 5.0, 5.3, 5.8, 6.0, 5.5～6.2, 4.6,5.1,5.5～5.8 respectively. The identification ratios of the sperm membrane proteins on 78 kD (60.7%), 60 kD (71.4%), 51 kD (14.9%) and 23 kD (14.29%) were higher. Conclusion The sperm membrane proteins with MW of 78 kD, 60 kD, 51 kD and 23 kD were associated with antisperm antibody and immunological infertility. Two- dimensional gel electrophoresis and Western blotting can precisely identify the sperm membrane proteins that are associated with antisperm antibody.
基金supported by Outstanding Scholars of New Era from Ministry of Education of Peopde's Republic of China(NECT-07-0861)the Lotus Scholars Program of Hunan Province, P.R. China (2007-362)Scientific and Technological Projects of Department of Public Health of Hunan, P.R. China (B2009123)
文摘目的:筛选人急性髓系白血病细胞株HL-60甲基化沉默基因,为揭示白血病的发病机制及防治提供科学依据。方法:应用荧光差异双向凝胶电泳(two-dimensional fluorescence difference gel electro-phoresis,F-2D-DIGE)分离甲基转移酶抑制剂5-杂氮-2-脱氧胞苷(5-aza-2-dC)处理与未处理的HL-60细胞的总蛋白质,Decyder和PDquest图像分析软件识别差异表达蛋白质点,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)技术鉴定差异表达蛋白质。结果:建立了5-aza-2-dC处理与未处理的HL-60细胞蛋白质的F-2D-DIGE图谱,图像分析识别了53个差异表达的蛋白质点,质谱分析鉴定了35个差异表达的蛋白质,其中32个蛋白质在5-aza-2-dC处理后的HL-60细胞中表达上调,3个蛋白质表达下调。结论:35个差异表达蛋白可能与甲基化相关,为白血病的表观遗传学研究提供了有价值的信息。
基金Supported by Natural Science Fund for Colleges and Universities in Anhui Province, China (No.KJ2007B208)
文摘AIM: To establish and compare serum proteomic of diabetic retinopathy (DR) patients in various phases and discuss pathogenesis of DR so as to find out possible serum specific molecular markers for early diagnosis of DR. METHODS: Thirty-two subjects were divided into four groups: one group of eight type 2 diabetes mellitus (T2DM) patients without apparent DR (No-DR, NDR), one group of eight T2DM patients with non-proliferative diabetic retinopathy (NPDR), one group of eight T2DM patients with proliferative diabetic retinopathy(PDR) and one group of eight healthy volunteer participants. Two dimensional fluorescence difference gel electrophoresis (2D-DIGE) was applied to establish differential protein expression profiles in four groups. Matrix-assisted laser desorption/ionization time of flight tandem mass spectrometry (MALDI-TOF-TOF MS) was applied to identify mass spectrometry of differential proteins and analyze follow-up bioinformatics. RESULTS: 2D-DIGE maps of serum protein were satisfactory obtained from NDR, NPDR, PDR and normal control groups. Twenty-six different proteins spots were screened(the volume ratio was > 1.5 based on DeCyder software analysis). Twenty-four of them were verified and two of them were not. Fifteen proteins were verified. Most of them were high-abundant proteins in serum. The four relatively low-abundant ones were beta 2-glycoprotein I (beta (2)-GPI), alpha2-HS-glycoprotein (AHSG), alpha1-acid glycoprotein (alpha(1)-AGP) and apolipoprotein A-1 (apo A-1). beta (2)-GPI expression was gradually increased in the development of DR but unrelated to the severity of DR. The volume ratio of beta (2)-GPI is 1.54, 2.43, and 2.84 in NDR, NPDR and PDR group respectively compared with normal control group. CONCLUSION: Serum proteomic analysis of 2D-DIGE combined with MALDI-TOF-TOF MS is feasible to be applied in the study of DR. 13 2-GPI probably takes part in the process of DR occurrence and development and it could be a candidate biomarker on DR diagnosis in early phase.
基金sponsored by a grant from the Sichuan Provincial Education Department(Influence of AGS3 on"reversible protein phosphorylation"in cortical neu-rons of morphine-addicted rats),No.2005B016
文摘The prefrontal cortex is involved in the regulation and control of substance addiction-related cognitive, behavioral, and emotional changes. The present study identified prefrontal cortex protein profiles in morphine-addicted rats; these were subsequently compared with normal rats. Results showed 87 protein spots with differentially expressed levels in the morphine addiction group, with the majority located in meta acid zones at pH 4.2-6.8 and having a molecular weight of 30-110 kDa In addition, 2 protein spots were identified as being associated with neurotoxicity （Snap25 isoform β-Snap25 of synaptosomal-associated protein 25 and βactin）.
文摘目的:探讨双向差异凝胶电泳(two dimension difference gel electrophoresis,2D-DIGE)的实验方法,分析比较其优缺点,概述其在微生物研究中的应用。方法:以两株不同血清型的副溶血性弧菌为样本,通过2D-DIGE实验,得到两组蛋白质荧光染色分布图,与银染的效果比较。结果:直观的看到两株细菌的蛋白质差异,且可分析样品丰度变化。结论:该方法有一定的局限性,但却是研究蛋白质组丰度变化的强有力工具。
