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1.
目的比较黄曲霉毒素B1(AFB1)诱发的树鼩肝细胞癌生长不同阶段的蛋白质表达谱的改变,筛查在AFB1诱发的肝癌发生发展中起重要作用的关键蛋白质分子.方法用双向电泳(2-DE)及基质辅助激光解吸飞行时间串联质谱法(MALDI-TOF-MS/MS)对同一动物自身的癌发生前肝组织(A45W),癌发生后的肝癌组织(ACa)及未经AFB1处理的对照组动物肝组织(E45W)进行了相互比较.结果A45W组、ACa组、E45W组分别平均检测到(1 255±43)个蛋白点(n=3)、(1 198±50)个蛋白点(n=3),和(1 252±40)个蛋白点(n=3),以其中A45W组图谱为参考胶,ACa组、E45W组分别与其匹配的平均匹配点数为(818±39)个(n=3)和(777±45)个(n=3),平均匹配率分别为66%和62%.癌与癌前比(ACa vs A45W),相差2倍以上的上调点和下调点分别为108个(包括癌特异点)和75个;癌前与未经AFB1处理的对照组比(A45W vs E45W),相差2倍以上的上调点有78个,下调点有21个.质谱可鉴定出不均一核糖核蛋白A1、不均一核糖核酸核蛋白A2/B1、peroxiredoxin 1、peroxiredoxin 2、膜联蛋白4、肌球蛋白α链等26种胶内差异蛋白质.结论在AFB1诱发的树鼩肝癌发生不同阶段有明显的差异性蛋白表达谱,为进一步对人肝癌蛋白质组的研究提供了线索.  相似文献   

2.
目的解析比较蛋白质组学筛出的差异蛋白S100A4的生物学功能,验证其是否在肝癌转移复发中发挥作用。方法用逆转录聚合酶链反应(RTPCR)、蛋白免疫印迹(WB)及细胞免疫化学法分别对差异蛋白S100A4在转移潜能不同人肝癌细胞系中的表达情况进行再验证;然后,用构建S100A4反义表达质粒,转染高转移潜能人肝癌细胞系MHCC97H,通过系列与侵袭转移有关的检测,观察S100A4表达降低对MHCC97H细胞生物学行为的影响。结果验证结果均证实,S100A4在有转移潜能人肝癌细胞系MHCC97L、MHCC97H中呈高表达。转染S100A4反义重组表达质粒后的侵袭试验显示,穿过人工基底膜细胞数分别是:5.0±1.4[MHCC97H/pcDNA3.1(+)ASS100A4],16.4±1.6[MHCC97H/pcDNA3.1(+)],16.9±1.5(MHCC97H);运动试验穿过上室底膜的细胞数为:11.1±3.3[MHCC97H/pcDNA3.1(+)ASS100A4],18.9±2.0[MHCC97H/pcDNA3.1(+)],21.9±3.4(MHCC97H);细胞培养上清液中MMP9的定量结果依次为18.2[MHCC97H/pcDNA3.1(+)ASS100A4]、28.0[MHCC97H/pcDNA3.1(+)]、31.9ng/ml(MHCC97H);MMP2定量结果依次为29.8[MHCC97H/pcDNA3.1(+)ASS100A4]、26.4[MHCC97H/pcDNA3.1(+)]、26.5ng/ml(MHCC97H)。明胶酶谱分析细胞培养上清液基质金属蛋白酶(MMP)显示,S100A4表达降低的MHCC97H细胞分泌的MMP9活性明显弱于对照组。结论pcDNA3.1(+)ASS100A4转染MHCC97H细胞后,细胞的运动与侵袭潜能均明显降低。细胞MMP9分泌量也显著降低。而细胞分泌MMP2的量则变化不明显。提示差异蛋白S100A4通过增强肝癌细胞的运动侵袭能力和上调MMP的分泌参与肝癌细胞的侵袭转移过程。  相似文献   

3.
目的:胎兔皮肤蛋白质组的研究可能解开其无瘢痕愈合的机制,建立重复性好、分辨率高的胎兔皮肤组织蛋白质组双向电泳技术,初步分析寻找皮肤切割伤伤后胎兔蛋白质表达的差异。方法:本实验于2004-01/12在第三军医大学野战外科研究所完成。取孕20d的新西兰大耳白兔、行宫内手术致胎兔皮肤全层切割伤,24h后取致伤胎兔及其同胞未致伤胎兔背部皮肤,分别确定为致伤组和未致伤组,应用哺乳动物总蛋白提取试剂盒提取其总蛋白。应用双向电泳法将致伤组和未致伤组分别制成3块固相pH梯度胶条,电泳结束后硝酸银染色、图像扫描。同一胶条进行3次双向电泳,对所得图像进行蛋白质点、匹配率以及伤后蛋白表达差异的分析。结果:致伤组和未致伤组胎兔皮肤组织总蛋白分别分离出蛋白质点(105±14)个、(108±19)个,在两组中各选1块胶作为参考胶进行3块胶间的匹配,平均匹配点数分别是(88±8)个、(90±7)个,匹配率分别为84%、83%,同一样本3块胶上不同蛋白质点在第一向的位置偏差分别为(0.85±0.25)mm、(0.9±0.4)mm,在第二向的偏差分别为(1.25±0.5)mm、(1.5±0.5)mm。通过比对得到胎兔伤后表达增加的蛋白质点20个。结论:加大皮肤组织总蛋白上样量进行双向电泳得到重复性和分辨率均较好的电泳图谱,并初步找到伤后差异表达的蛋白质点  相似文献   

4.
C反应蛋白在冠心病及经皮冠状动脉介入治疗中意义   总被引:1,自引:0,他引:1  
于立成  王伟民  师淑敏  沙亚美  刘键  吴淳 《临床荟萃》2004,19(19):1093-1096
目的 探讨在稳定型心绞痛 (SA) ,不稳定型心绞痛 (UA)和冠状动脉造影正常组三组病例C反应蛋白(C reactiveprotein ,CRP)水平变化的临床意义。探讨CRP变化与手术参数的相关性。探讨CRP在住院期间预测主要不良心脏事件 (majoradversecardiacevent,MACE)的价值。方法 测定入选患者 (n =97)CRP水平 ,观察冠状动脉性心脏病 (CHD)病例中高CRP水平组 (n =2 5 )及低CRP水平组 (n =35 )围手术期及住院期间MACE。结果 UA组 (n=37)CRP水平高于SA(n=2 7)和冠状动脉造影组 (n =33) [(2 3.4± 2 .3)mg/Lvs(6 .1± 8.3)mg/L ,P <0 .0 1;(2 3.4± 2 .3)mg/Lvs(4 .6± 3.7)mg/L ,P <0 .0 0 1]。经皮冠状动脉介入治疗 (percutaneouscoronaryintervention ,PCI)组 (n =39)手术后CRP水平明显增高 (P <0 .0 0 1) ,经皮穿刺冠状动脉球囊成型术 (percutaneoustransluminalcoronaryangioplasty,PTCA)总时间 >10秒与PTCA总时间≤ 10秒CRP手术前后差值差异有统计学意义 [(2 8.2±17.6 )mg/L vs(17.2± 7.1)mg/L ,P <0 .0 5 ],支架最大扩张压力 ,支架扩张总时间 ,支架长度两组间CRP手术前后差值差异均无统计学意义。高CRP组 (CRP >15mg/L)CHD围手术期及住院期间反复发作心绞痛高于低CRP组(CRP <15mg/L) (P <0 .0 5 )。结论 血清  相似文献   

5.
邱俊  周继红  张波 《中国临床康复》2005,9(14):56-57,i002
目的:胎兔皮肤蛋白质组的研究可能解开其无瘢痕愈合的机制,建立重复性好、分辨率高的胎兔皮肤组织蛋白质组双向电泳技术,初步分析寻找皮肤切割伤伤后胎兔蛋白质表达的差异.方法:本实验于2004-01/12在第三军医大学野战外科研究所完成.取孕20 d的新西兰大耳白兔、行宫内手术致胎兔皮肤全层切割伤,24 h后取致伤胎兔及其同胞未致伤胎兔背部皮肤,分别确定为致伤组和未致伤组,应用哺乳动物总蛋白提取试剂盒提取其总蛋白.应用双向电泳法将致伤组和未致伤组分别制成3块固相pH梯度胶条,电泳结束后硝酸银染色、图像扫描.同一胶条进行3次双向电泳,对所得图像进行蛋白质点、匹配率以及伤后蛋白表达差异的分析.结果:致伤组和未致伤组胎兔皮肤组织总蛋白分别分离出蛋白质点(105&;#177;14)个、(108&;#177;19)个,在两组中各选1块胶作为参考胶进行3块胶间的匹配,平均匹配点数分别是(88&;#177;8)个、(90&;#177;7)个,匹配率分别为84%、83%,同一样本3块胶上不同蛋白质点在第一向的位置偏差分别为(0.85&;#177;0.25)mm、(0.9&;#177;0.4)mm,在第二向的偏差分别为(1.25&;#177;0.5)mm、(1.5&;#177;0.5)mm.通过比对得到胎兔伤后表达增加的蛋白质点20个.结论:加大皮肤组织总蛋白上样量进行双向电泳得到重复性和分辨率均较好的电泳图谱,并初步找到伤后差异表达的蛋白质点,为无瘢痕愈后的深入实验研究打下基础.  相似文献   

6.
Objective To investigate the plasma 8-iso-prostaglandin F2α(8-iso-PGF2α and the serum C-re-active protein(CRP) levels in patients with obstructive sleep apnea-hypopnea syndrome(OSAHS) with and without hypertension(OSAHS + HT),and to explore the changes of pathophysiology in patients with OSAHS and the patho-genesis of OSAHS + HT. Methods All observed subjects were divided into 3 groups: control group (n=20),OS-AHS group(n=19),OSAHS + HT group (n=21). Plasma 8-iso-PGF2αand serum CRP concentrations levels were measured by ELISA and were compared. Results The plasma 8-iso-PGF2αand serum CRP levels,were higher in OSAHS patients than those in control subjects [(11.08±3.26)μg/L vs (7.49±2.10)μg/L,P<0.01;(1.75±0.82) mg/L vs (0.52±0.26 ) mg/L,P<0.01],and were higher in OSAHS + HT group than those in control group [14.84±3.43)μG/L vs(11.08±3.26)μg/L,P<0.01 ;(3.13±1.06)mg/L vs(1.75±0.82)mg/L,P<0.01]. Conclusions Oxidative stress and inflammation in OSAHS patients are increased,which are involved in the devel-opment of OSAHS associated hypertension.  相似文献   

7.
Objective To investigate the plasma 8-iso-prostaglandin F2α(8-iso-PGF2α and the serum C-re-active protein(CRP) levels in patients with obstructive sleep apnea-hypopnea syndrome(OSAHS) with and without hypertension(OSAHS + HT),and to explore the changes of pathophysiology in patients with OSAHS and the patho-genesis of OSAHS + HT. Methods All observed subjects were divided into 3 groups: control group (n=20),OS-AHS group(n=19),OSAHS + HT group (n=21). Plasma 8-iso-PGF2αand serum CRP concentrations levels were measured by ELISA and were compared. Results The plasma 8-iso-PGF2αand serum CRP levels,were higher in OSAHS patients than those in control subjects [(11.08±3.26)μg/L vs (7.49±2.10)μg/L,P<0.01;(1.75±0.82) mg/L vs (0.52±0.26 ) mg/L,P<0.01],and were higher in OSAHS + HT group than those in control group [14.84±3.43)μG/L vs(11.08±3.26)μg/L,P<0.01 ;(3.13±1.06)mg/L vs(1.75±0.82)mg/L,P<0.01]. Conclusions Oxidative stress and inflammation in OSAHS patients are increased,which are involved in the devel-opment of OSAHS associated hypertension.  相似文献   

8.
Objective To investigate the plasma 8-iso-prostaglandin F2α(8-iso-PGF2α and the serum C-re-active protein(CRP) levels in patients with obstructive sleep apnea-hypopnea syndrome(OSAHS) with and without hypertension(OSAHS + HT),and to explore the changes of pathophysiology in patients with OSAHS and the patho-genesis of OSAHS + HT. Methods All observed subjects were divided into 3 groups: control group (n=20),OS-AHS group(n=19),OSAHS + HT group (n=21). Plasma 8-iso-PGF2αand serum CRP concentrations levels were measured by ELISA and were compared. Results The plasma 8-iso-PGF2αand serum CRP levels,were higher in OSAHS patients than those in control subjects [(11.08±3.26)μg/L vs (7.49±2.10)μg/L,P<0.01;(1.75±0.82) mg/L vs (0.52±0.26 ) mg/L,P<0.01],and were higher in OSAHS + HT group than those in control group [14.84±3.43)μG/L vs(11.08±3.26)μg/L,P<0.01 ;(3.13±1.06)mg/L vs(1.75±0.82)mg/L,P<0.01]. Conclusions Oxidative stress and inflammation in OSAHS patients are increased,which are involved in the devel-opment of OSAHS associated hypertension.  相似文献   

9.
Objective To investigate the plasma 8-iso-prostaglandin F2α(8-iso-PGF2α and the serum C-re-active protein(CRP) levels in patients with obstructive sleep apnea-hypopnea syndrome(OSAHS) with and without hypertension(OSAHS + HT),and to explore the changes of pathophysiology in patients with OSAHS and the patho-genesis of OSAHS + HT. Methods All observed subjects were divided into 3 groups: control group (n=20),OS-AHS group(n=19),OSAHS + HT group (n=21). Plasma 8-iso-PGF2αand serum CRP concentrations levels were measured by ELISA and were compared. Results The plasma 8-iso-PGF2αand serum CRP levels,were higher in OSAHS patients than those in control subjects [(11.08±3.26)μg/L vs (7.49±2.10)μg/L,P<0.01;(1.75±0.82) mg/L vs (0.52±0.26 ) mg/L,P<0.01],and were higher in OSAHS + HT group than those in control group [14.84±3.43)μG/L vs(11.08±3.26)μg/L,P<0.01 ;(3.13±1.06)mg/L vs(1.75±0.82)mg/L,P<0.01]. Conclusions Oxidative stress and inflammation in OSAHS patients are increased,which are involved in the devel-opment of OSAHS associated hypertension.  相似文献   

10.
Objective To investigate the plasma 8-iso-prostaglandin F2α(8-iso-PGF2α and the serum C-re-active protein(CRP) levels in patients with obstructive sleep apnea-hypopnea syndrome(OSAHS) with and without hypertension(OSAHS + HT),and to explore the changes of pathophysiology in patients with OSAHS and the patho-genesis of OSAHS + HT. Methods All observed subjects were divided into 3 groups: control group (n=20),OS-AHS group(n=19),OSAHS + HT group (n=21). Plasma 8-iso-PGF2αand serum CRP concentrations levels were measured by ELISA and were compared. Results The plasma 8-iso-PGF2αand serum CRP levels,were higher in OSAHS patients than those in control subjects [(11.08±3.26)μg/L vs (7.49±2.10)μg/L,P<0.01;(1.75±0.82) mg/L vs (0.52±0.26 ) mg/L,P<0.01],and were higher in OSAHS + HT group than those in control group [14.84±3.43)μG/L vs(11.08±3.26)μg/L,P<0.01 ;(3.13±1.06)mg/L vs(1.75±0.82)mg/L,P<0.01]. Conclusions Oxidative stress and inflammation in OSAHS patients are increased,which are involved in the devel-opment of OSAHS associated hypertension.  相似文献   

11.
目的对肝细胞肝癌(hepatocellular carcinoma,HCC)患者尿液代谢轮廓进行分析,建立疾病区分模型并筛选出具有潜在诊断价值的特征代谢标志物。方法采用超高效液相色谱与质谱联用代谢组学研究平台对27。例HCC患者,31例肝硬化患者及22例健康志愿者尿液样本进行分析,利用Mzmine2.7软件对数据进行数据前处理及归一化;利用SIMCA—P+12.0.1.0软件进行主成分分析(prinei-pal component analysis,PCA)及正交偏最小二乘(orthogonal partial leastsquares discriminant analysis,OPLS—DA)聚类分析,构建疾病区分模型;筛选特征代谢物,并利用SPSS17.0软件进行数据统计分析。结果构建了HCC与肝硬化及健康对照尿液代谢轮廓的PCA(R2X:66.9%,Q2=35.4%)及OPLS—DA疾病区分模型(R2X=59.4%,R2Y=92.1%,Q2=82.8%),并且筛选18个对比HCC组与肝硬化组或健康对照组存在显著差异的特征离子,并鉴定了其中的15个。3葡萄糖苷、四氢醛固酮3葡萄糖甘酸等15种特征代谢离子在HCC组与肝硬化组差异均有统计学意义(P均〈0.05),5-a-双氢睾酮葡糖苷酸等12种特征离子在HCC组与健康对照组间差异均有统计学意义(P均〈0.05)。结论利用代谢轮廓分析方法所找到的尿液中特征代谢物具有很好的区分HCC患者与肝硬化及健康人群的能力,可以作为潜在肝脏疾病的诊断标志物及其新的治疗靶点,在临床领域可做进一步研究。  相似文献   

12.
目的:研究宫颈鳞癌患者血清差异蛋白质与鳞状细胞癌抗原(squamous cell carcinoma antigen,SCCA)之间的关系,寻找早期诊断宫颈癌的生物学指标。方法:采用表面增强激光解吸离子化飞行时间质谱仪(SELDI-TOF-MS)和弱阳离子结合芯片(CM10),检测44例宫颈鳞状细胞癌及25例健康女性的血清蛋白质,将3组SCCA不同水平的宫颈癌患者分别与健康对照组进行对比,同时通过对比SCCA不同水平的宫颈癌患者,筛选出有分类意义的差异蛋白质。结果:(1)宫颈鳞癌SCCA正常患者与健康对照组比较,共有19种血清蛋白质质谱峰值差异具有显著性(P<0.001)。宫颈鳞癌SCCA升高患者(大于正常1倍)以及大于正常3倍组,分别与健康对照组比较,分别发现了26种和15种差异蛋白质(P<0.05)。其中有4种质荷比(M/Z)为4172、3397、2741、6086的差异蛋白质,重复出现于宫颈鳞癌SCCA正常或升高不同水平组与健康对照组的比较中。(2)在宫颈鳞癌患者中,SCCA小于正常3倍组与大于正常3倍组比较、SCCA小于正常4倍组与大于正常4倍组比较,分别发现了5种和20种差异蛋白质(P<0.05)。其中M/Z为8142、7765、3885、5632、9287的差异蛋白质,重复出现于上述两组比较中。结论:通过蛋白芯片-飞行质谱方法,能够更早的发现宫颈癌与正常人之间的血清差异蛋白质,这些差异蛋白质有潜力发展为宫颈癌早期诊断指标,有助于宫颈癌的筛查、区分早晚期宫颈癌以指导治疗,具有重要的临床应用价值。  相似文献   

13.
BACKGROUND: Recent advances in proteomic profiling technologies, such as surface-enhanced laser desorption/ionization mass spectrometry (SELDI), have allowed preliminary profiling and identification of tumor markers in biological fluids in several cancer types and establishment of clinically useful diagnostic computational models. We developed a bioinformatics tool and used it to identify proteomic patterns in urine that distinguish transitional cell carcinoma (TCC) from noncancer. METHODS: Proteomic spectra were generated by mass spectroscopy (surface-enhanced laser desorption and ionization). A preliminary "training" set of spectra derived from analysis of urine from 46 TCC patients, 32 patients with benign urogenital diseases (BUD), and 40 age-matched unaffected healthy men were used to train and develop a decision tree classification algorithm that identified a fine-protein mass pattern that discriminated cancer from noncancer effectively. A blinded test set, including 38 new cases, was used to determine the sensitivity and specificity of the classification system. RESULTS: The algorithm identified a cluster pattern that, in the training set, segregated cancer from noncancer with sensitivity of 84.8% and specificity of 91.7%. The discriminatory pattern correctly identified. A sensitivity of 93.3% and a specificity of 87.0% for the blinded test were obtained when comparing the TCC vs. noncancer. CONCLUSIONS: These findings justify a prospective population-based assessment of proteomic pattern technology as a screening tool for bladder cancer in high-risk and general populations.  相似文献   

14.

Objectives

Nowadays, nearly 1000 hemoglobin (Hb) variants are known. The standard biochemical techniques used in Hb analysis are mainly: isoelectric focusing, cation-exchange liquid chromatography (LC) and reversed-phase LC. In addition to this approach, a protein analysis is achieved by mass spectrometry (MS) and additional DNA studies are performed. The aim of this review is to emphasize the significance of MS methods applied to Hb variants analysis.

Results and perspectives

To perform Hb studies, different MS techniques are currently used such as electrospray ionization (ESI), matrix-assisted laser desorption ionization (MALDI) and tandem mass spectrometry (MS/MS). As shown here, MS is an efficient tool for identification of all types of variants (substitution of a single amino acid residue, several substitutions in the same globin chain, insertions/deletions, fusion Hbs). The use of MS in neonatal screening of Hb variants is also presented.

Conclusions

MS is a powerful technique for Hb analysis. It appears as being an important additional method in the set of biochemical techniques.  相似文献   

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目的建立食管鳞癌诊断预测模型,寻找食管鳞癌潜在肿瘤标志物。方法收集59例食管鳞癌患者及57例体检健康者血清标本,按照3∶1随机分为训练组(患者44例,健康人对照42例)和验证组(患者15例,健康人对照15例)。弱阳离子交换磁珠(WCX-MB)提纯血清蛋白质/肽后,经基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)检测,筛选出差异表达蛋白质/肽,构建诊断预测模型并初步验证。结果经Clin ProTools软件分析,共筛选出差异多肽峰31个(P0.05),其中18个峰差异有统计学意义(P0.01);与健康人对照组相比,8个峰在食管鳞癌组中表达上调; 10个峰在食管鳞癌组中表达下调。其中差异峰m/z 2 660.84和m/z 5 336.49的ROC曲线下面积(AUCROC)分别为0.95和0.91,且均在食管鳞癌组中表达上调。应用遗传算法(GA)建立的食管鳞癌诊断预测模型经验证其准确性为93.10%,敏感性为92.90%,特异性为93.30%。结论建立的食管鳞癌诊断预测模型能较好的用于食管鳞癌辅助诊断,多肽峰m/z 2 660.84和m/z 5 336.49有望成为食管鳞癌潜在肿瘤标志物。  相似文献   

16.
PTEN在胃癌中的表达   总被引:1,自引:0,他引:1  
目的检测FIEN基因在胃癌组织中的表达,并研究其临床病理意义。方法采用逆转录聚合酶链式反应(RT-PCR)法检测FFEN在40例胃癌中的表达。结果①PINM分期Ⅰ、Ⅱ期胃癌与对照组正常胃黏膜组织FFEN的表达有显著性差异(P〈0.01);FTNM分期Ⅲ、Ⅳ期和Ⅰ、Ⅱ期胃癌FFEN的表达有非常显著性差异(P〈0.05);②FIEN在高、中分化腺癌和低分化胃癌阳性率表达差异具有显著性(P〈0.05)。③FFEN在有淋巴结转移组和无淋巴结转移组检出阳性率差异具有显著性(P〈0.05)。结论FFEN的表达与胃癌临床病理特征和生物学行为存在密切关系,FFEN基因是胃癌细胞增生活跃的重要影响因素。  相似文献   

17.
目的检测MMP-2、PTEN基因在胃癌组织中的表达,并研究其临床病理意义。方法采用逆转录聚合酶链式反应(RT-PCR)法检测MMP-2、PTEN在40例胃癌中的表达。结果①PTNM分期Ⅰ、Ⅱ期胃癌与对照组正常胃黏膜组织MMP-2、PTEN的表达均有显著性差异(P<0·01);PTNM分期Ⅲ、Ⅳ期和Ⅰ、Ⅱ期胃癌MMP-2、PTEN的表达均有非常显著性差异(P<0·05);②MMP-2、PTEN在高、中分化腺癌和低分化胃癌阳性率表达差异均具有显著性(P<0·05)。③MMP-2、PTEN在有淋巴结转移组和无淋巴结转移组检出阳性率差异具有显著性(P<0·05)。结论MMP-2、PTEN的表达与胃癌临床病理特征和生物学行为存在密切关系,MMP-2、PTEN基因是胃癌细胞增生活跃的重要影响因素。  相似文献   

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