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皮肤细胞蛋白质组质谱分析方法的建立及肽质量指纹图谱的构建
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作者 陈斌 毕志刚 《中国中西医结合皮肤性病学杂志》 CAS 2006年第3期125-131,共7页
目的建立人皮肤细胞蛋白质组学研究所需的质谱分析方法,并获得皮肤细胞蛋白质肽质量指纹图谱。方法体外培养人皮肤成纤维细胞与角质形成细胞,采用固相pH梯度双向凝胶电泳分离其总蛋白质,凝胶经染色后,在图谱上选取2个角质形成细胞蛋白点... 目的建立人皮肤细胞蛋白质组学研究所需的质谱分析方法,并获得皮肤细胞蛋白质肽质量指纹图谱。方法体外培养人皮肤成纤维细胞与角质形成细胞,采用固相pH梯度双向凝胶电泳分离其总蛋白质,凝胶经染色后,在图谱上选取2个角质形成细胞蛋白点与1个成纤维细胞蛋白点,进行胶内酶切、基质辅助激光解吸电离飞行时间(MALDI-TOF)质谱分析鉴定。结果获得3个点的肽质量指纹图谱,经Mascot软件搜索人非冗余蛋白质数据库后,鉴定为细胞骨架Actingamma1、tubulinα2及热休克蛋白HSP70。结论皮肤细胞基质辅助激光解吸电离飞行时间(MALDI-TOF)质谱分析技术的建立为皮肤蛋白质组学的进一步研究提供手段。 展开更多
关键词 蛋白质组 双向凝胶电泳 角质形成细胞 成纤维细胞 基质辅助激光解吸电离飞行时间质谱 肽质量指 纹图谱
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HIV感染导致MT4细胞蛋白表达差异的初步研究 被引量:1
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作者 李林 应天翼 +9 位作者 庄道民 鲍作义 刘思扬 李韩平 刘永健 杨坤 李钰 王恒良 黄留玉 李敬云 《中国病毒学》 CSCD 2004年第5期435-438,共4页
为了比较MT4细胞株感染HIV-1的ⅢB株前后的蛋白质表达差异,我们分别提取MT4细胞及感染了人类免疫缺陷病毒(HIV)的MT4细胞的总蛋白质,通过双向电泳分离,使用ImageMaster2DElite3.10图像分析软件分析获得的凝胶图谱,寻找差异点,使用质谱... 为了比较MT4细胞株感染HIV-1的ⅢB株前后的蛋白质表达差异,我们分别提取MT4细胞及感染了人类免疫缺陷病毒(HIV)的MT4细胞的总蛋白质,通过双向电泳分离,使用ImageMaster2DElite3.10图像分析软件分析获得的凝胶图谱,寻找差异点,使用质谱仪鉴定获得的差异点蛋白质。结果表明感染HIV和未感染HIV的MT4细胞有40个蛋白质点差异,HIV感染后减少的蛋白质点有12个,增多的有28个,通过质谱分析,29个蛋白质得到鉴定。其中HIV感染后下调的蛋白质有能量代谢相关蛋白、肌动蛋白相关蛋白及假想蛋白等;上调的蛋白有肌动蛋白、酶类蛋白、免疫蛋白及假想蛋白等。通过研究我们可以看出宿主细胞感染HIV病毒后有多个蛋白发生变化,可能和HIV与宿主细胞的相互作用有关。为了研究HIV感染的机制必须去除高丰度蛋白,针对特定功能的蛋白质进行具体研究。 展开更多
关键词 MT4 人免疫缺陷病毒(Human IMMUNODEFICIENCY virus HIV):双向电泳:肽质量指文谱分析
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Diagnostic model of saliva protein finger print analysis of patients with gastric cancer 被引量:9
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作者 Zheng-Zhi Wu Ji-Guo Wang Xiao-Li Zhang 《World Journal of Gastroenterology》 SCIE CAS CSCD 2009年第7期865-870,共6页
AIM:To explore the method for early diagnosis of gastric cancer by screening the expression spectrum of saliva protein in gastric cancer patients using mass spectrometry for proteomics.METHODS:Proportional peptide mas... AIM:To explore the method for early diagnosis of gastric cancer by screening the expression spectrum of saliva protein in gastric cancer patients using mass spectrometry for proteomics.METHODS:Proportional peptide mass fingerprints were obtained by analysis based on proteomics matrix-assisted laser desorption ionization time-of-flight/mass spectrometry.A diagnosis model was established using weak cation exchange magnetic beads to test saliva specimens from gastric cancer patients and healthy subjects.RESULTS:Significant differences were observed in the mass to charge ratio(m/z) peaks of four proteins(1472.78 Da,2936.49 Da,6556.81 Da and 7081.17 Da) between gastric cancer patients and healthy subjects.CONCLUSION:The finger print mass spectrum of saliva protein in patients with gastric cancer can be established using gastric cancer proteomics.A diagnostic model for distinguishing protein expression mass spectra of gastric cancer from non-gastric-cancer saliva can be established according to the different expression of proteins 1472.78 Da,2936.49 Da,6556.81 Da and 7081.17 Da.The method for early diagnosis of gastric cancer is of certain value for screening special biological markers. 展开更多
关键词 SALIVA Protein finger print model Gastriccancer Matrix-assisted laser desorption ionization-time-of-flight/mass spectrometry Weak cationexchange Magnetic bead PROTEOMICS
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Optimization of DsbA Purification from Recombinant Escherichia coli Broth Using Box-Behnken Design Methodolog 被引量:1
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作者 LUO Man GUAN Yixin YAO Shanjing 《Chinese Journal of Chemical Engineering》 SCIE EI CAS CSCD 2013年第2期185-191,共7页
Disulfide bond formation protein A (DsbA) is one of the important helper proteins for folding in protein synthesis in vivo. In this study, purification of recombinant DsbA was investigated by examining four importan... Disulfide bond formation protein A (DsbA) is one of the important helper proteins for folding in protein synthesis in vivo. In this study, purification of recombinant DsbA was investigated by examining four important factors with Box-Behnken design method, a statistic-based design of experiments. The optimal operation conditions were obtained by adopting the effectiveness coefficient method on the multi-objective problem, which takes the protein recovery, purification efficiency and throughput of ion-exchange chromatography into account. After the optimization, protein recovery of 96.8% and purity higher than 95% DsbA was achieved, and the productivity was (377.9±1.7) mg soluble DsbA per liter broth. The purified protein was identified by peptide mass fingerprinting matching the record of gil2624856, a mutant of DsbA. The DsbA was preliminarily applied to the refolding of denatured lysozyme in vitro. 展开更多
关键词 disulfide bond formation protein A protein purification Box-Behnken experiment design response surface methodology multi-object programming
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Murine brain tissues with human cytomegalovirus infection: a proteomic study
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作者 Xie Ni Yuan Jianhui +1 位作者 Cai Yinsha Liu Jianjun 《Journal of Medical Colleges of PLA(China)》 CAS 2010年第2期65-74,共10页
To establish two-dimensional electrophoresis profiles with high resolution and reproducibility from murine brain tissues by human cytomegalovirus(HCMV) infection and paired murine brain tissues and to identify the d... To establish two-dimensional electrophoresis profiles with high resolution and reproducibility from murine brain tissues by human cytomegalovirus(HCMV) infection and paired murine brain tissues and to identify the differential expression proteins. Methods: Forty Kunming mice were randomly divided into infection group (20) injected with HCMVAD169 and control group (20) injected with saline into their brain. After 30 days, the murine brain tissues by HCMV infection and paired murine brain tissues were separated by two-dimensional gel electrophoresis(2-DE), analyzed by Image Master 2D software, and identified by peptide mass fingerprint(PMF) and database searching, and make Western blotting analyses the differential expression of individual proteins. Results: Well resolved, reproducible 2-D maps of the above tissues were obtained. Some of the different proteins identified by mass spectrometry(MS) were matched in the SWISS-2D PAGE database, Western blotting analyses were further carried out to verify the differential expression of individual proteins. Conclusion: These data will be valuable for studying the diagnosis of disease at an early stage, mechanisms of pathogenic and the key to the development of anti-HCMV medicine. 展开更多
关键词 Human cytomegalovirus INFECTION PROTEOMICS Differential expression
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Effect of acupuncture on serum PYY and nesfatin-1 in obese patients with insulin resistance 被引量:1
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作者 Liu Mei-zhang Liu Yong-tao Yang Li-bai 《Journal of Acupuncture and Tuina Science》 CSCD 2021年第6期432-437,共6页
Objective To observe the effect of acupuncture on serum peptide YY(PYY)and nesfatin-1 in obese patients with insulin resistance.Methods Ninety-eight obese patients with insulin resistance were divided into a control g... Objective To observe the effect of acupuncture on serum peptide YY(PYY)and nesfatin-1 in obese patients with insulin resistance.Methods Ninety-eight obese patients with insulin resistance were divided into a control group and an observation group by the random number table method,with 49 cases in each group.The control group received exercise and dietary interventions,and the observation group received additional acupuncture treatment to the exercise and dietary interventions.The body mass index(BMI),body fat percentage,fasting insulin(FINS),fasting plasma glucose(FPG),homeostasis model assessment for insulin resistance(HOMA-IR),triglyceride(TG),total cholesterol(TC),and serum PYY and nesfatin-1 levels were compared before and after treatment.The efficacy was evaluated after treatment.Results The total effective rate was significantly higher in the observation group than in the control group(P<0.05).After treatment,the BMI,body fat percentage,and serum TG and TC levels decreased significantly in both groups(P<0.05),and were significantly lower in the observation group than those in the control group(P<0.05).The FINS,FPG,HOMA-IR,and serum PYY and nesfatin-1 levels of the control group were not significantly changed after treatment(P>0.05).The FINS,FPG and HOMA-IR of the observation group decreased significantly after treatment(P<0.05),and were lower than those in the control group(P<0.05).The serum PYY and nesfatin-1 levels of the observation group increased significantly after treatment(P<0.05),and were higher than those in the control group(P<0.05).Conclusion Based on exercise and dietary interventions,acupuncture is effective for obese patients with insulin resistance.It can reduce the BMI,body fat percentage,blood lipids,blood glucose,and serum insulin levels and improve insulin resistance.The action may be associated with the up-regulation of serum PYY and nesfatin-1. 展开更多
关键词 Acupuncture Therapy OBESITY Diet Reducing Insulin Resistance Body Mass Index Peptide YY NESFATIN-1
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