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IDENTIFICATION OF DIFFERENTIAL PROTEINS IN UTERINE LEIOMYOMA BY TWO-DIMENSIONAL ELECTROPHORESIS
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作者 朱雪琼 朱春丹 +1 位作者 呂杰强 董克 《Chinese Journal of Cancer Research》 SCIE CAS CSCD 2006年第3期203-208,共6页
Objective: To establish and optimize the two-demensional electrophoresis maps of uterine leiomyoma and to study the difference of global protein patterns between uterine leiomyoma and normal myometrium. Methods: Usi... Objective: To establish and optimize the two-demensional electrophoresis maps of uterine leiomyoma and to study the difference of global protein patterns between uterine leiomyoma and normal myometrium. Methods: Using Two-dimensional electrophoresis followed by computer-assisted image analysis, the differential proteins between uterine leiomyoma and normal myometrium were compared. Results: The well-resolved and reproducible two-dimensional gel electrophoresis patterns of uterine leiomyoma and normal myometrium were established. Totally 1085±108 and 1103±151 protein spots were obtained by using the pH 4-7 IPG strips in uterine leiomyoma and normal myometrium map, respectively, of which 7 spots increased and 15 spots decreased in quantity in uterine leiomyoma compared with normal myometrium. Conclusion: The differentially expressed proteins are useful for studying the mechanism of the cause of uterine leiomyoma. 展开更多
关键词 two-dimensional electrophoresis Uterine leiomyoma PROTEOME differential expression
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Two-dimensional Electrophoresis Analysis of Differential Protein Expression in Squamous Carcinoma of the Cervix
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作者 朱雪琼 吴洁丽 +4 位作者 余丽蓉 林毅 吕杰强 邹双微 胡越 《Chinese Journal of Cancer Research》 SCIE CAS CSCD 2008年第3期164-170,共7页
Objective: To establish and optimize the two-dimensional gel electrophoresis (2-DE) maps of squamous carcinoma of the cervix and to study the protein difference between squamous carcinoma of the cervix (SCC) and ... Objective: To establish and optimize the two-dimensional gel electrophoresis (2-DE) maps of squamous carcinoma of the cervix and to study the protein difference between squamous carcinoma of the cervix (SCC) and normal cervical tissue. Methods: Using Two-dimensional gel electrophoresis followed by computer-assisted image analysis, the differential proteins between squamous carcinoma of the cervical tissue and normal cervical tissue were compared. Then using matrix-assisted laser desorption/ionization-time of flight mass spectrometry, the differential proteins were identified. Results: The well-resolved and reproducible two-dimensional gel electrophoresis patterns of squamous carcinoma of the cervix tissue and normal cervical tissue were obtained. After silver staining, the average matching ratio of squamous carcinoma of the cervix was 86.1%. There was a good reproducibility of spot position in 2-DE map, with average deviation in IEF direction of 0.95±0.13 mm, while in SDS-PAGE direction it was 1.20±0.18 mm. Ten protein spots were identified by mass spectrometry, some of which were involved in cell proliferation, cell apoptosis, intracellular enzymes, structural proteins, cycle regulation, and tumor occurrence. Conclusion: The differentially expressed proteins provide a fundamental basis for further study of human squamous carcinoma of the cervix and screening of its specific markers. 展开更多
关键词 two-dimensional gel electrophoresis Cervical cancer PROTEOME differential expression
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Protein expression of sensory and motor nerves Two-dimensional gel electrophoresis and mass spectrometry
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作者 Zhiwu Ren Yu Wang +5 位作者 Jiang Peng Li Zhang Wenjing Xu Xiangdang Liang Qing Zhao Shibi Lu 《Neural Regeneration Research》 SCIE CAS CSCD 2012年第5期369-375,共7页
The present study utilized samples from bilateral motor branches of the femoral nerve, as well as saphenous nerves, ventral roots, and dorsal roots of the spinal cord, to detect differential protein expression using t... The present study utilized samples from bilateral motor branches of the femoral nerve, as well as saphenous nerves, ventral roots, and dorsal roots of the spinal cord, to detect differential protein expression using two-dimensional gel electrophoresis and nano ultra-high performance liquid chromatography electrospray ionization mass spectrometry tandem mass spectrometry techniques. A mass spectrum was identified using the Mascot search. Results revealed differential expression of 11 proteins, including transgelin, Ig kappa chain precursor, plasma glutathione peroxidase precursor, an unnamed protein product (gil55628), gfyceraldehyde-3-phosphate dehydrogenase-like protein, lactoylgfutathione lyase, adenyfate kinase isozyme 1, two unnamed proteins products (gil55628 and gi11334163), and poly(rC)-binding protein 1 in motor and sensory nerves. Results suggested that these proteins played roles in specific nerve regeneration following peripheral nerve injury and served as specific markers for motor and sensory nerves. 展开更多
关键词 differential protein expression mass spectrometry motor nerve peripheral nerve-specific regeneration two-dimensional gel electrophoresis sensory nerve
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Proteome analysis of round-headed and normal spermatozoa by 2-D fluorescence difference gel electrophoresis and mass spectrometry 被引量:9
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作者 Ting-Ting Liao Zhen Xiang Wen-Bing Zhu Li-Qing Fan 《Asian Journal of Andrology》 SCIE CAS CSCD 2009年第6期683-694,共12页
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
关键词 differential protein GLOBOZOOSPERMIA mass spectrometry (MS) two-dimensional difference gel electrophoresis (2-D DIGE)
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Differential Gene and Protein Expression in Soybean at Early Stages of Incompatible Interaction with Phytophthora sojae 被引量:1
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作者 LI Yong-gang YANG Ming-xiu +3 位作者 LI Yan LIU Wen-wen WEN Jing-zhi LI Yong-hao 《Agricultural Sciences in China》 CAS CSCD 2011年第6期902-910,共9页
Soybean root and stem rot caused by Phytophthora sojae is a destructive disease worldwide. Using genetic resistance is an important and major component in the integrated pest management of this disease. To understand ... Soybean root and stem rot caused by Phytophthora sojae is a destructive disease worldwide. Using genetic resistance is an important and major component in the integrated pest management of this disease. To understand molecular mechanisms of root and stem rot resistance in soybeans, the gene and protein expression in hypocotyls and stems of variety Suinong 10 carrying resistance genes Rps1a and Rps2 was investigated by using mRNA differential display reverse transcription PCR and two-dimensional electrophoresis at 0, 0.5, 1, 2, and 4 h after inoculation with P. sojae race 1. The results of the comparison of gene and protein expression showed that at least eight differential fragments at the transcriptional level were related to metabolic pathway, phytoalexin, and signal transduction in defense responses. Sequence analyses indicated that these fragments represented cinnamic acid 4-hydroxylase gene, ATP b gene coding ATP synthase b subunit and ubiquitin-conjugating enzyme gene which upregulated at 0.5 h post inoculation, blue copper protein gene and UDP-N-acetyl-a-D-galactosamine gene which upregulated at 2 h post inoculation, TGA-type basic leucine zipper protein TGA1.1 gene, cyclophilin gene, and 14-3-3 protein gene which upregulated at 4 h post inoculation. Three resistance-related proteins, a-subunit and b-subunit of ATP synthase, and cytochrome P450-like protein, were upregulated at 2 h post inoculation. The results suggested that resistance-related multiple proteins and genes were expressed in the recognition between soybean and P. sojae during zoospore germination, penetration and mycelium growth of P. sojae in soybean. 展开更多
关键词 Phytophthora sojae resistance mechanism incompatible interaction mRNA differential display reverse transcription PCR two-dimensional electrophoresis
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Two-dimensional gel electrophoresis analysis of the proteomes expressed in the human hepatoma cell line BEL-7404 and normal liver cell line L-02 被引量:1
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作者 YU Lirong WANG Nan +2 位作者 WU Gaode XU Yonghua XIA Qichang 《Chinese Science Bulletin》 SCIE EI CAS 2000年第12期1113-1122,共10页
Proteome analysis technology has been used extensively in conducting discovery research of biology and has become one of the most essential technologies in functional genomics. The proteomes of the human hepatoma cell... Proteome analysis technology has been used extensively in conducting discovery research of biology and has become one of the most essential technologies in functional genomics. The proteomes of the human hepatoma cell line BEL-7404 and the normal human liver cell line L-02 have been separated by high resolution two-dimensional gel electrophoresis (2-DE) with immobilized pH gradient isoelectric focusing (IPG-IEF) in the first dimension and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) in the second dimension (IPG-DALT). The resulting images have been analyzed using 2-D analysis software. Quantitative analysis reveals that 7 protein spots are detected only in hepatoma BEL-7404 cells, 14 only in L-02 cells, and 78 protein spots show significant fluctuation in quantity in both cell lines (P【0.01). These protein spots have been displayed on a proteome differential expression map. Analysis for the reproducibility of 2-DE indicates that the positional variability in the IEF dimension 展开更多
关键词 PROTEOME two-dimensional gel electrophoresis differential expression map human HEPATOMA CELLS liver cells.
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Characterization of Acute Renal Allograft Rejection by Human Serum Proteomic Analysis 被引量:2
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作者 高英 吴轲 +9 位作者 徐逸 周鸿敏 何文涛 张维娜 蔡兰军 林星光 方泽民 雒真龙 郭晖 陈忠华 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2009年第5期585-591,共7页
To identify acute renal allograft rejection biomarkers in human serum, two-dimensional differential in-gel electrophoresis (2-D DIGE) and reversed phase high-performance liquid chromatography (RP-HPLC) followed by... To identify acute renal allograft rejection biomarkers in human serum, two-dimensional differential in-gel electrophoresis (2-D DIGE) and reversed phase high-performance liquid chromatography (RP-HPLC) followed by electrospray ionization mass spectrometry (ESI-MS) were used. Serum samples from renal allograft patients and normal volunteers were divided into three groups: acute rejec- tion (AR), stable renal function (SRF) and normal volunteer (N). Serum samples were firstly processed using Multiple Affinity Removal Column to selectively remove the highest abundance proteins. Differentially expressed proteins were analyzed using 2-D DIGE. These differential protein spots were excised, digested by trypsin, and identified by RP-HPLC-ESI/MS. Twenty-two differentially expressed proteins were identified in serum from AR group. These proteins included complement C9 precursor, apolipoprotein A-IV precursor, vitamin D-binding protein precursor, beta-2-glycoprotein 1 precursor, etc. Vitamin D-binding protein, one of these proteins, was confirmed by ELISA in the independent set of serum samples. In conclusion, the differentially expressed proteins as serum biomarker candidates may provide the basis of acute rejection noninvasive diagnosis. Confirmed vitamin D-binding protein may be one of serum biomarkers of acute rejection. Furthermore, it may provide great insights into understanding the mechanisms and potential treatment strategy of acute rejection. 展开更多
关键词 acute rejection two-dimensional differential in-gel electrophoresis reversed phase high-performance liquid chromatography electrospray ionization mass spectrometry ELISA SERUM
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Proteomic Analysis of Proteins in the Salivary Glands of the Fed and Unfed Female Tick Rhipicephalus haemaphysaloides
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作者 XIANG Fei-yu ZHANG Jian-wu +3 位作者 ZHOU Yong-zhi LI Zhuang GONG Hai-yan ZHOU Jin-lin 《Agricultural Sciences in China》 CAS CSCD 2009年第1期121-127,共7页
Identification of differentially expressed salivary gland proteins between the fed and unfed female Rhipicephalus haemaphysaloides may obtain valuable functional molecules. In this study, comparative two-dimensional g... Identification of differentially expressed salivary gland proteins between the fed and unfed female Rhipicephalus haemaphysaloides may obtain valuable functional molecules. In this study, comparative two-dimensional gel electrophoresis (2-DE) and mass spectrometry were used to separate and identify differentially expressed salivary gland proteins between the fed and unfed female R. haemaphysaloides. The soluble proteins from the salivary glands of fed and unfed female R. haemaphysaloides were separated by a sequential extraction method followed by 2-DE and 2-DE images. Image analysis of the gels revealed 1 096 ± 87 protein spots from the fed female ticks and 991 ±64 protein spots from the unfed female ticks. Among those protein spots, about 724 ±34 were present both in the fed and unfed female ticks. Fourteen spots from the fed ticks and six spots from the unfed ticks were selected for peptide mass fingerprinting (PMF) and sequencing assay by mass spectrometry (MS). Bioinformatic analysis showed that a majority of the differentially expressed proteins were involved in signal transduction, metabolism, and transcriptional regulation. These differentially expressed proteins might be antigen candidates for the development of vaccines against the tick. 展开更多
关键词 Rhipicephalus haemaphysaloides salivary g/and differentially expressed proteins two-dimensional gel electrophoresis mass spectrometry
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Original article Preliminary analysis of cerebrospinal fluid proteome in patients with neurocysticercosis 被引量:5
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作者 TIAN Xiao-jun LI Jing-yi +1 位作者 HUANG Yong XUE Yan-ping 《Chinese Medical Journal》 SCIE CAS CSCD 2009年第9期1003-1008,共6页
Background Neurocysticercosis is the infection of the nervous system by the larvae of Taenia solium (T. solium). Despite continuous effort, the experimental diagnosis of neurocysticercosis remains unresolved. Since ... Background Neurocysticercosis is the infection of the nervous system by the larvae of Taenia solium (T. solium). Despite continuous effort, the experimental diagnosis of neurocysticercosis remains unresolved. Since the cerebrospinal fluid (CSF) contacts with the brain, dynamic information about pathological processes of the brain is likely to be reflected in CSF. Therefore, CSF may serve as a rich source of putative biomarkers related to neurocysticercosis. Comparative proteomic analysis of CSF of neurocysticercosis patients and control subjects may find differentially expressed proteins.Methods Two-dimensional difference in gel electrophoresis (2D-DIGE) was used to investigate differentially expressed proteins in CSF of patients with neurocysticercosis by comparing the protein profile of CSF from neurocysticercosis patients with that from control subjects. The differentially expressed spots/proteins were recognized with matrix-assisted laser desorption/ionization-time of flight-time of flight (MALDI-TOF-TOF) mass spectrometry.Results Forty-four enzyme digested peptides were obtained from 4 neurocysticercotic patients. Twenty-three were identified through search of the NCBI protein database with Mascot software, showing 19 up-expressed and 4 down-expressed. Of these proteins, 26S proteosome related to ATP- and ubiquitin-dependent degradation of proteins and lipocalin type prostaglandin D synthase involved in PGD2-synthesis and extracellular transporter activities were up-expressed, while transferrin related to iron metabolism within the brain was down-expressed.Conclusions This study established the proteomic profile of pooled CSF from 4 patients with neurocysticercosis, suggesting the potential value of proteomic analysis for the study of candidate biomarkers involved in the diagnosis or pathogenesis of neurocysticercosis. 展开更多
关键词 NEUROCYSTICERCOSIS cerebrospinal fluid two-dimensional difference in-gel electrophoresis oroteomic analysis
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Quantitative proteomics analysis of parthenogenetically induced pluripotent stem cells
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作者 Zhe Hu LeiWang +9 位作者 Zhensheng Xie Xinlei Zhang Du Feng Fang Wang Bingfeng Zuo Lingling Wang Zhong Liu Zhisheng Chen Fuquan Yang Lin Liu 《Protein & Cell》 SCIE CSCD 2011年第8期631-646,共16页
Parthenogenetic embryonic stem(pES)cells isolated from parthenogenetic activation of oocytes and embryos,also called parthenogenetically induced pluripotent stem cells,exhibit pluripotency evidenced by both in vitro a... Parthenogenetic embryonic stem(pES)cells isolated from parthenogenetic activation of oocytes and embryos,also called parthenogenetically induced pluripotent stem cells,exhibit pluripotency evidenced by both in vitro and in vivo differentiation potential.Differential proteomic analysis was performed using differential in-gel electrophoresis and isotope-coded affinity tag-based quantitative proteomics to investigate the molecular mechanisms underlying the developmental pluripotency of pES cells and to compare the protein expression of pES cells generated from either the in vivo-matured ovulated(IVO)oocytes or from the in vitro-matured(IVM)oocytes with that of fertilized embryonic stem(fES)cells derived from fertilized embryos.A total of 76 proteins were upregulated and 16 proteins were downregulated in the IVM pES cells,whereas 91 proteins were upregulated and 9 were downregulated in the IVO pES cells based on a minimal 1.5-fold change as the cutoff value.No distinct pathways were found in the differentially expressed proteins except for those involved in metabolism and physiological processes.Notably,no differences were found in the protein expression of imprinted genes between the pES and fES cells,suggesting that genomic imprinting can be corrected in the pES cells at least at the early passages.The germline competent IVM pES cells may be applicable for germ cell renewal in aging ovaries if oocytes are retrieved at a younger age. 展开更多
关键词 parthenogenetic embryonic stem cell PROTEOME fluorescent two-dimensional difference in-gel electrophoresis isotope-coded affinity tag
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