反义真核细胞转化生长因子βⅠ型受体与反义基质金属蛋白酶组织抑制因子-1联合作用对大鼠肝纤维化的影响

目的观察反义转化生长因子βⅠ型受体（TβRI真核表达质粒与反义基质金属蛋白酶组织抑制因子-1（TIMP-1）真核表达质粒联合作用对实验性大鼠肝纤维化的影响。方法构建大鼠反义真核细胞表达质粒，导入大鼠肝纤维化模型体内，通过I型胶原的免疫组织化学以及苦味酸-酸性品红染色观察两种反义质粒联合作用对大鼠肝纤维化的影响，用目标积分吸光度（A）值表示各实验组动物肝组织中蛋白的表达量。结果反义TβRI疗组、反义TβRI反义TIMP-1治疗组、反义TIMP-1治疗组、pcDNA3．1（＋）空质粒对照组、模型对照组、正常对照组TβRI白的A值分别为：（2．11±0．88）×10^5、（1．06±0．57）×10^5、（3．46±1．14）×10^5、（5．66±2．54）×10^5、（5．19±1．22）×10^5和（0．38±0．27）×10^5；TIMP-1蛋白的A值分别为：（1．10±0．22）×10^5、（0．30±0．12）×10^5、（0．65±0．15）×10^5、（2．05±0．36）×10^5、（1．97±0．28）×10^5和（0．10±0．12）×10^5；I型胶原的蛋白表达的A值分别为：（4．37±1．30）×10，、（0．90±0．32）×10^5、（3．40±0．91）×10^5、（6．90±1．61）×10^5、（7．34±1．68）×10^5和（0．41±0．21）×10s。反义TIMP-1治疗组TIMP-1蛋白表达量显著降低（P〈0．05），反义TβRI疗组TβRI白表达量显著降低（P〈0．05），两种反义质粒可有效抑制相应蛋白的表达；反义TIMP-1表达质粒与反义TβRI达质粒均可减少受损肝脏中I型胶原的沉积（P〈0．05），联合应用可进一步减少受损肝脏中I型胶原的沉积（P〈0．01）。在病理形态学方面的观察，反义TIMP-1表达质粒与反义TβRI达质粒均可使受损肝脏的病理形态有一定改善，联合应用可使受损肝脏的病理形态得到进一步的改善。结论反义TIMP-1表达质粒与反义TβRI达质粒对肝纤维化的发展均有一定的干预作用，联合作用可产生更有效的阻止作用。     

慢性乙肝患者肝组织及血清中HBV DNA与肝脏组织病变的相关性

采用定量聚合酶链反应（FQ-PCR）检测45例慢性乙肝患者肝组织与血清中乙肝病毒（HBV）DNA的含量,并与丙氨酸氨基转移酶（ALT）及肝组织病理的关系进行相关性分析。结果 肝组织及血清中HBV DNA含量分别为2.8×10^5～1.1×109copies/cm3及2.4×10^3～8.1×10^7copies/cm3,P〈0.001,二者含量呈显著正相关（r=0.86,P〈0.001）。肝组织HBV DNA含量与ALT水平、肝组织炎症活动度及肝组织纤维化指标均无明显相关性。认为HBV复制程度与肝细胞损伤程度无明显相关性;肝组织中HBV DNA水平能更准确反映HBV的复制程度;血清中HBV DNA水平一定程度上可指导抗病毒治疗及判断疗效。     

Survivin在原发性肝细胞癌中的表达及意义

目的　Survivin是凋亡抑制蛋白中的一种 ,选择性地表达于恶性肿瘤组织。该文研究Sur vivin基因在原发性肝细胞癌中的表达及生物学意义。方法　收集 2株肝细胞癌细胞株 ,4 0例原发性肝癌组织标本及相应的癌旁组织 ,以Westernblotting法检测Survivin蛋白表达 ;半定量RT PCR法检测SurvivinmRNA表达 ;肝癌细胞凋亡指数采用原位末端标记法检测。结果　 2株肝癌细胞株和 85 % (34例 )的肝癌组织表达Survivin蛋白和mRNA ,而癌旁组织内无一例阳性表达。Survivin蛋白表达的阳性率在肝内转移组为 93.5 % ,显著高于肝内无转移组 (5 5 .6 % ,P <0 .0 5 ) ;在门静脉癌栓浸润组为 92 .8% ,显著高于无门静脉癌栓浸润组 (6 6 .7% ,P <0 .0 5 )。RT PCR显示 ,2株肝癌细胞株和 85 .0 % (34例 )的肝癌组织表达SurvivinmRNA ,与Westernblotting的结果一致 ,SurvivinmRNA的表达水平在肝内转移组 (1.10 5± 0 .396 )和门静脉癌栓浸润组 (1.137± 0 .4 0 4 )中 ,显著高于肝内无转移组 (0 .5 72± 0 .0 82 )和无门静脉癌栓浸润组 (0 .6 2 7± 0 .12 2 ,P <0 .0 5 )。所有肝癌组织标本中均可检测到凋亡细胞 ,但Survivin表达阳性组的凋亡指数 (1.15 2 %± 0 .32 6 % )显著低于Survivin表达阴性组 (4.5 0 2 %± 0 .830 % ,P <0 .0 5     

41例肝细胞癌门静脉癌栓患者64层螺旋CT门静脉成像诊断体会

目的 探讨64层CT门静脉成像(64-SCTP)的诊断门静脉癌栓的价值.方法 对41例肝细胞癌(HCC)患者行64-SCTP,并以最大密度投影法(MIP)、容积再现(VR)和多平面重建(MPR)技术进行图像后处理. 结果 发现门静脉主干癌栓15例,右支癌栓11例,左支癌栓5例,主干+左支癌栓2例,主干+右支癌栓8例,门静脉海绵样变11例;CT轴位图像诊断准确门静脉癌栓40例(7.8％),MPR结合MIP诊断41例(00％),二者差异无统计学意义.结论 64-SCTP可多方位显示门静脉癌栓部位及范围.     

血管紧张素Ⅱ1型受体拮抗剂对胰腺癌细胞的抑制作用

目的探讨选择性血管紧张素Ⅱ1型受体拮抗剂ZD7155在体外对胰腺癌细胞PaTu8988的抑制作用及其作用机制。方法采用四甲基偶氮唑盐（MTT）法测定浓度为5×10^-11、5×10^-10。、5×10^9、5×10^-8、5×10^-7和5×10^6mol／L的ZD7155在同一时间点和5×10^-8mol／LZD7155在不同作用时点对人胰腺癌细胞系PaTu8988的影响，应用流式细胞仪检测ZD7155作用前后PaTu8988细胞周期变化，电镜观察ZD7155对PaTu8988细胞凋亡的影响以及ZD7155对PaTu8988细胞形态的影响。结果MTT显示ZD7155的抑制率随着浓度增大、时间延长而增加。浓度分别为5×10^-11、5×10^10、5×10^-9、5×10^-8、5×10^-7和5×10^6mol／L的ZD7155在与PaTu8988细胞作用48h后，对其的抑制率分别为9％、18％、30％、51％、60％和78％。相同浓度（5．0×10^-8mol／L）的ZD7155在与PaTu8988作用后12、24、36、48、60和72h对其抑制率分别为15％、25％、36％、51％、67％和85％。ZD7155对人胰腺癌细胞系PaTu8988细胞周期S期有明显抑制作用，而且呈剂量依赖性。经与ZD7155共同孵育的细胞电镜下可见细胞核染色质边集、凋亡小体形成，而且随浓度升高而愈明显，细胞形态学未发生改变。结论ZD7155在体外能抑制胰腺癌细胞生长和增殖，其作用机制可能与其抑制细胞周期S期以及诱导细胞凋亡有关，且无明显细胞毒性作用。     

门静脉癌栓影像学诊断的比较

比较术前各影像学检查对门静脉癌栓诊断的敏感性和特异性。回顾性分析110例肝细胞肝癌(HCC)伴门静脉癌栓患者的术前影像学特征及诊断。以同期100例HCC不伴门静脉癌栓患者为阴性对照。US、CT和MRL／MRA诊断门静脉癌栓的敏感性分别为88%、、93％和94％,其特异性分别为95％、96％和98％。经x2检验,各影像学诊断门静脉癌栓敏感性和特异性的差异无统计学意义(P>0．05)。各影像学检查对门静脉癌栓诊断的敏感性和特异性相近。     

双途径化疗治疗肝癌并发门静脉癌栓患者对预后的影响

目的探讨经药物传输泵(DDS)门静脉化疗(PVC)联合肝动脉插管化疗栓塞(TACE)]治疗肝细胞癌伴门静脉癌栓的临床疗效及安全性。方法对87例肝细胞癌合并门静脉癌栓患者手术切除肿瘤并取出癌栓,再随机将患者分成3组。A组行TACE、B组行经DDS泵PVC,C组行经DDS泵PVC联合TACE。随访3年。结果 C组患者术后6个月、1年、2年和3年无瘤生存率分别为93.5%、80.6%、41.9%和29.0%,累积生存率分别为96.8%、87.1%、54.8%和45.2%,其中2年和3年生存率显著高于A组和B组(P均<0.05);C组患者术后6个月、1年、2年和3年上消化道出血发生率分别为0%、0%、16.1%和22.6%,与A组和B组比无显著性差异(P均>0.05)。结论对肝细胞癌合并门静脉癌栓患者行手术切除肿瘤并取出癌栓后,行双途径化疗可有效提高患者无瘤生存率及累计生存率。     

重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白对慢性阻塞性肺疾病大鼠肺功能的影响

目的探讨重组人Ⅱ型肿瘤坏死因子受体一抗体融合蛋白（rhTNFR：Fc）对大鼠慢性阻塞性肺疾病（COPD）模型肺功能的影响。方法48只大鼠随机分为正常对照组、COPD组、rhTNFR：Fc干预组（简称干预组）及假干预组。单纯吸烟法建立COPD模型，干预组皮下注射rhTNFR：Fc进行干预，假干预组皮下注射空白模拟制剂。酶联免疫吸附法（ELISA）测定各组血清和支气管肺泡灌洗液（BALF）中的肿瘤坏死因子α（TNF—α）浓度。肺组织切片行苏木精一伊红染色观察形态学改变，定量测定肺平均内衬间隔和平均肺泡数，用小动物肺功能仪测定系统测定肺功能。结果COPD组0．3秒用力呼气容积占用力肺活量比值（FEV。／FVC）和呼气峰流速（PEF）[（65．1±8．4）％和（18．801．6）ml／s]显著低于正常对照组[（85．6±5．9）％和（47．2±7．3）mt／s]和干预组[（77．7±2．7）％和（38．2±3．3）mL／s]，假干预组FEV。／FVC和PEF[（65．4±9．8）％和（19．0-1-1．9）mL／s]显著低于干预组。COPD组血清中TNF-α浓度[（118±34）ng／L]显著高于正常组[（74±16）ng／L]和干预组[（79±14）ng／L]，假干预组血清中TNF—α浓度[（120±39）ng／L]显著高于干预组；COPD组BALF中TNF—α浓度[（155±28）ng／L]显著高于正常对照组[（79±28）ng／L]。COPD组肺平均内衬间隔[（77±29）×10^-6m／个]显著高于正常对照组[（44±7）×10^-6m／个]，其平均肺泡数[（232±97）×10^6个／m^2]显著低于正常对照组[（393±24）×10^6个／m^2]和干预组[（379±33）×10^6个／m^2]。假干预组平均肺泡数[（213±99）×10^6个／m^2]显著低于干预组[（379±33）×10^6个／m^2]。COPD组＋假干预组血清和BALF中TNF—α浓度与PEF和FEV。／FVC均呈显著负相关。结论TNF—α与吸烟大鼠的COPD形成有关，并影响其肺功能；rhTNFR：Fc对延缓肺功能损害具有一定的作用。     

菜菔硫烷对低硒病毒性心肌损伤小鼠Ⅱ相酶和一氧化氮的影响

目的 研究Ⅱ相酶和一氧化氮（NO）在低硒病毒性心肌损伤中的变化以及十字花科蔬菜成分菜菔硫烷（SF）的影响。方法 采用低硒和常硒病毒性心肌损伤小鼠模型，观察心肌组织Ⅱ相酶和NO的变化及SF对其的影响。结果 病毒感染使低硒条件下谷胱甘肽硫转移酶（GST）水平增高[（22．46±2．08）×10^3、（19．36±2．03）×10^3U／g，P〈0．05]，而常硒条件下使醌还原酶（QR）水平增高[（24．40±1．18）×10^3、（22．04±1．25）×10^3U／g，P〈0．01]，各SF保护组Ⅱ相酶活性均明显高于相应未保护组（P〈0．05）；病毒感染使小鼠血清NO水平升高（P〈0．01）。常硒SF保护组NO水平显著低于未保护组[（45．98±3．16）、（61．33±8．54）μmol／L，P〈0．01]。结论 SF可通过诱导心肌组织Ⅱ相酶活性．提高抗氧化能力以及降低血清NO水平减轻炎症反应来保护损伤的心肌。     

新生大鼠肝细胞脾内移植联合鼠肝再生增强因子治疗大鼠急性肝衰竭的疗效与免疫学机制

目的探讨新生大鼠肝细胞脾内移植联合重组鼠肝再生增强因子（rALR）治疗大鼠急性肝衰竭的疗效与免疫学机制。方法采用D-氨基半乳糖（1．2 g／kg）诱导并建立大鼠急性肝衰竭模型,36 h后随机分为6组：Ⅰ组：模型对照组;Ⅱ组：经脾脏注射等渗盐水1 ml,腹腔注射等渗盐水1 ml／d;Ⅲ组：经脾脏注射rALR 1 ml（50μg／kg）,腹腔注射rALR 1 ml（50μg·kg^-1·d^-1）;Ⅳ：经脾脏移植2×10^7同种异体新生大鼠肝细胞,腹腔注射等渗盐水1 ml／d;Ⅴ组：经脾脏移植2×10^7同种异体新生大鼠肝细胞,腹腔注射rALR 1 ml（50μg·kg^-1·d^-1）;Ⅵ组：经脾脏移植2×10^7同种异体新生大鼠肝细胞,腹腔注射环孢菌素1 ml（10 mg·kg^-1·d^-1）。观察大鼠每天存活率;术后第1、5天及2周处死大鼠以获取肝、脾组织,观察脾内移植肝细胞的组织学改变;在术前、术后第1、2、5、12天采集静脉血,采用放射免疫法测定血清IL-1β和TNFα的浓度。结果Ⅰ、Ⅱ和Ⅲ组大鼠存活时间差异无统计学意义,Ⅳ组有部分大鼠长期存活,2周存活率为33％,Ⅴ组大鼠2周存活率显著高于Ⅳ组,而与Ⅵ组大鼠2周存活率比较差异无统计学意义。Ⅳ、Ⅵ组脾内移植的肝细胞可存活5～7 d,Ⅴ组脾内移植肝细胞可存活至2周以上。术后第1天,Ⅳ组血清IL-1β水平显著高于Ⅴ组和Ⅵ组（P〈0．05）;术后第5天,Ⅳ组血清IL-1β水平仅与Ⅵ组差异有统计学意义（P〈0．05）。术后第1天,TNFα的浓度在Ⅱ组显著高于Ⅳ、Ⅴ、Ⅵ组（P〈0．05）。结论新生大鼠肝细胞脾内移植与rALR联合治疗大鼠急性肝衰竭有一定疗效,可提高D-氨基半乳糖诱导肝衰竭大鼠的存活率;移植肝细胞在脾脏可存活2周以上,其机制可能与促进肝细胞再生、预防移植肝细胞凋亡及轻微抑制细胞免疫有关。     

Screening and detection of portal vein tumor thrombi-associated serum low molecular weight protein biomarkers in human hepatocellular carcinoma

Purpose Serum low molecular weight protein biomarkers might be important in relation to portal vein tumor thrombi (PVTT) in hepatocellular carcinoma (HCC). This study aimed to screen and to detect these biomarkers. Methods We selected sera of 3 groups from 12 healthy volunteers, 12 HCC patients without PVTT and 12 HCC patients with PVTT, respectively. By using two-dimensional gel electrophoresis (2-DE) in which the first dimension was 16% SDS-PAGE, serum protein images of 3 groups were analyzed by Image Master Software. The differential protein spots were further identified by MALDI-TOF MS/MS. Results Compared with 12.5% SDS-PAGE gel, there were more protein bands between 3 and 20 kDa in 16% SDS-PAGE gel and low molecular weight (MW) protein spots (<20 kDa) were clearly shown. Fifteen differential protein spots representing five proteins were found in the three groups by inter-class comparison and were then identified. Compared with the healthy group, apolipoprotein A-I, lipoprotein CIII, transthyretin and DNA topoisomerase II were down regulated in HCC groups while haptoglobin-2 was over expressed. All the five proteins were less in PVTT group than in non-PVTT group. Conclusion The expression of low MW serum protein changes obviously in the beginning and progressive stage of HCC, and differentially expressed low MW proteins might be the potential biomarkers in early prognostication and surveillance of treatment for HCC and PVTT.     

Identification of two portal vein tumor thrombosis associated proteins in hepatocellular carcinoma: protein disulfide-isomerase A6 and apolipoprotein A-I

Background and Aim: Portal vein tumor thrombus (PVTT) is one of the factors that can affect prognosis and survival of hepatocellular carcinoma (HCC). In the present study, we aimed to find out some biomarkers associated with vascular invasion features of HCC with the method of comparative proteomic analysis. Methods: The proteins were extracted from a pair of HCC tissues with PVTT and without PVTT, and then separated by two‐dimensional polyacrylamide gel electrophoresis. Differentially expressed protein spots were identified by matrix assisted laser desorption ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS). Further analysis of two proteins were completed using real‐time fluorescence quantitative polymerase chain reaction and western‐blot in 40 HCC tissues with PVTT (n = 20) and without PVTT (n = 20). Results: Among 465 protein spots displayed on the gels, 33 unique proteins (> twofold change, P < 0.01) were identified, including 24 upregulated in HCC tissue without PVTT and nine upregulated in HCC tissue with PVTT. The real‐time fluorescence quantitative PCR showed no statistically significant difference between HCC tissues with PVTT and without PVTT for mRNA expressions of protein disulfide‐isomerase, A6 (PDI A6) (P = 0.137) and apolipoprotein A‐I (Apo A‐I) (P = 0.718). However, compared with HCC tissues without PVTT, protein expression of PDI A6 was higher in HCC tissues with PVTT (P < 0.001), while protein expression of Apo A‐I was lower in HCC tissues with PVTT (P = 0.012). Conclusions: PDI A6 and Apo A‐I are closely related to vascular invasion feature of HCC.     

血清蛋白质组分析技术筛选肝癌自发抗体

目的采用血清蛋白质组分析技术(SERPA)筛选、鉴定肝癌自发抗体。方法双向电泳分离肝癌细胞系HCCLM3的总蛋白后将其转膜,肝癌、肝炎和正常组血清各8份与膜免疫印迹,图像分析确定不同血清免疫印迹图谱间的差异点及其与双向电泳图谱的对应关系,最后用基质辅助激光解吸飞行时间质谱进行鉴定。结果建立了高重复性HCCLM3的双向电泳图谱及其肝癌、肝炎及正常组血清的免疫印迹图谱,图谱均点数分别为603、70.75±24.25、68.50±23.44和41.38±15.05,肝癌、肝炎组图谱点数明显多于正常组,但肝癌与肝炎组差异无统计学意义。质谱鉴定确定了核蛋白、细胞骨架、代谢酶及热休克蛋白等五类肝癌自发抗体。结论SERPA是一种高通量筛选、鉴定肿瘤自发抗体的新技术,大量肝癌自发抗体的发现为肝癌进一步的免疫诊断及治疗奠定了基础。     

慢性砷暴露人群血清差异表达蛋白研究

目的 筛选慢性砷暴露人群血清差异表达蛋白,为寻找慢性砷暴露和砷中毒的血清蛋白标志物提供线索.方法 在山西省饮水型地方性砷中毒病区村进行现场调查,收集调查对象的人口学基本特征、饮水砷暴露史、吸烟、饮酒和健康情况等信息.根据饮水水砷浓度分组:低砷组(水砷暴露＜ 10μg/L)、中砷组(水砷暴露10 ～ 50 μg/L)、高砷组(水砷暴露＞50 μg/L)、砷中毒组(过去水砷暴露＞50 μg/L,改水后目前水砷＜10μg/L),选择符合入组条件的调查对象,每组30例.砷中毒组诊断依据《地方性砷中毒诊断标准》(WS/T 211-2001)进行.根据知情同意的原则,采集4组人群静脉血,取血清,采用双向差异凝胶电泳(two-dimensional differential gel electrophoresis,2-D DIGE)筛选不同水砷浓度暴露组及砷中毒组血清差异表达蛋白,采用质谱进行蛋白鉴定.结果 在6张胶图中平均检测出蛋白点(1299±167)个,其中在5张胶中均出现的蛋白点有688个.在低、中、高砷组,有33个蛋白点存在差异表达(P＜ 0.01);在低、中、高砷、砷中毒组,有54个蛋白点存在差异表达(P＜0.01).对其中的25个蛋白点进行了质谱鉴定,成功的鉴定出13种蛋白.与低砷组比较,载脂蛋白A-Ⅳ、血浆视黄醇结合蛋白、雌激素受体(下丘脑亚型)在中、高砷组表达下调,补体4A和4B表达上调;与低、中、高砷组比较,β2糖化蛋白Ⅰ、角蛋白1、血红素结合蛋白前体、补体C1r亚单位、纤维凝胶蛋白3在砷中毒组表达下调,色素上皮衍生因子、α1-微球蛋白、羧肽酶N端催化链表达上调.结论 慢性砷暴露对人群血清蛋白产生显著影响,砷中毒人群血清中出现的差异表达蛋白不会在短期内随着饮水砷浓度的下降而全部得到改善,鉴定的差异表达蛋白能否作为慢性砷暴露或砷中毒的血清标志物还有待于进一步验证.     

肝癌发生不同病理阶段血清蛋白质组的比较

目的 比较肝癌发生不同病理阶段血清蛋白质组的表达变化情况,从中寻找特异性标志物.方法 用双向凝胶电泳分离正常人,慢性肝炎、肝硬化和肝癌患者的血清蛋白质,结合质谱技术对差异点进行鉴定; Western blot检测结合珠蛋白β链以验证蛋白质水平的表达.计量资料数据以均数±标准差(x±s)表示,采用方差分析和LSD检验进行两两比较.结果 4组血清的双向电泳图谱经软件匹配分析,并与基质辅助激光解析电离飞行时间质谱相结合,成功鉴定出结合珠蛋白,SAA1、SP40等30个差异蛋白质点,K cluster聚类分析不同的变化模式.在差异蛋白质点中,7个蛋白质点均为结合珠蛋白,呈现由正常→肝炎→肝硬化持续下调、到肝癌又上调的模式.Western blot证实结合珠蛋白β链的表达与双向电泳表达相一致.结论 在肝癌发生的动态过程中各疾病阶段的血清蛋白质差异表达有4种明显变化模式,可能为肝癌的早期诊断和预后提供依据,与肝癌发生发展相关的结合珠蛋白很可能是肝硬化发展到肝癌的一个潜在早期诊断标志物.     

Comparative Proteomics of Sera From HCC Patients With Different Origins

Background:

Hepatocellular carcinoma (HCC), a major fatal cancer worldwide, is induced by different etiological factors in the liver.

Objectives:

To gain insight into serum protein profiling of HCC with different etiologies.

Patients and Methods:

We subjected the sera of HBV-HCC, HCV-HCC, non-B non-C-HCC patients, and healthy volunteers to two-dimensional gel electrophoresis (2-DE) and liquid chromatography tandem mass spectrometry (LC-MS/MS).

Results:

We found 30 differentially expressed protein spots (≥ 1.5 fold P < 0.05) between these two analyses; of them 17 protein spots corresponding to 8 proteins were identified by MS. Transthyretin, leucine rich α-2-glycoprotein, and ficolin 3 were differentially expressed between HBV-related HCC and non-B non-C-HCC sera. Moreover, haptoglobin α-2 isoforms were decreased in HCV-HCC compared to non-B non-CHCC.

Conclusions:

Serum proteome analyses of HCC with different origins showed a differential protein pattern, presumably related to different hepatopathogenesis in liver induced by different agents. Further studies are required to clarify the importance of identified proteins for early diagnosis of HCC with different origins.     

Plasma biomarkers of mouse aging

Normal aging is accompanied by a series of physiological changes such as gray hair, cataracts, reduced immunity, and increased susceptibility to disease. To identify novel biomarkers of normal aging, we analyzed plasma proteins of male mice longitudinally from 2 to 19 months of age. Plasma proteins were analyzed by two-dimensional gel electrophoresis and identified using mass spectrometry (MS), MS/MS and liquid chromatography MS/MS. We found that many plasma proteins exist as multiple isoforms with different masses and/or charges. Thirty-nine protein spots (corresponding to six distinct proteins) have been identified, 13 of which exhibited significant changes with age. For example, several proteins increased significantly during aging including one isoform of transthyretin, two isoforms of haptoglobin, and three isoforms of immunoglobulin kappa chain. Conversely, several proteins decreased significantly during aging including peroxiredoxin-2, serum amyloid protein A-1, and five isoforms of albumin. Identification of these proteins provides new biomarkers of normal aging in mice. If validated in humans, these biomarkers may facilitate therapeutic interventions to identify premature aging, delay aging, and/or improve healthspan of the elderly.     

不同转移潜能人肝癌细胞系酪氨酸磷酸化蛋白质差异分析

目的研究不同转移潜能人肝癌细胞系中酪氨酸磷酸化蛋白质表达的差异并筛查与肝癌转移相关的酪氨酸磷酸化蛋白质分子。方法应用双电泳(2-D E)、免疫印迹法及基质辅助激光解析电离飞行时间质谱分析,对三种不同转移潜能人肝癌细胞系Hep 3B、MHCC97L和MHCC97H进行酪氨酸磷酸化蛋白质组分析。结果对照2-DE胶和免疫印迹发光胶片,Hep3B检测到10个点,MHCC 9 7L检测到19个点, MHCC97H检测到17个点。经质谱鉴定,得到膜连蛋白Ⅰ等19个差异点。结论细胞内酪氨酸磷酸化蛋白的表达差异与肝癌的侵袭转移有关。     

Serum proteins in chronic hepatitis B patients treated with peginterferon alfa-2b

AIM: To study the differential protein profile in serum of hepatitis B patients.METHODS: Serum samples were obtained from patients with chronic hepatitis B who were receiving peginterferon alfa-2b.The serum samples were subjected to albumin depletion and analyzed by two-dimensional gel electrophoresis(2-DE).Differentially expressed protein spots were identified by electrospray ionizationquadrupole time-of-flight mass spectrometry.Alpha2-HS-glycoprotein,complement component C3c and CD5 antigen were further analyzed by an enzymelinked immunosorbent assay and immunonephelometry.RESULTS: Nineteen patients with HBeAg-positive chronic hepatitis B(CHB) were studied.These patients were followed for at least 1 year after treatment and were classified according to their treatment response: responders(n = 9) and non-responders(n = 10).2-DE and MS/MS analysis were performed to compare the serum proteins before initiating peginterferon alfa2b.From the quantitative analysis of the 2-D gel,7 proteins were detected between the two groups at different levels before treatment.Among these potential candidates,serum levels of alpha-2-HS-glycoprotein,complement component C3c and CD5 antigen-like precursor were further analyzed.In the validation phase,23 subjects,9 sustained responders and 14 nonresponders,were recruited.Interestingly,the levels of alpha-2-HS-glycoprotein and complement component C3c were elevated in the serum of the non-responders compared to the responders.CONCLUSION: Serum alpha-2-HS-glycoprotein and complement component C3c may be potential serum biomarkers in predicting the treatment response of peginterferon alfa-2b in patients with CHB prior to treatment.     

乙型肝炎病毒相关性肝细胞癌患者肝硬化组织与癌组织蛋白质组的差异

目的分析乙型肝炎病毒(HBV)相关性肝细胞癌患者肝硬化组织与癌组织蛋白质组之间的差异,鉴定其中的差异蛋白质点。方法应用双向聚丙烯酰胺凝胶电泳,对12例肝细胞癌患者的肝细胞癌组织与肝硬化组织的蛋白质进行差异分析,应用基质辅助激光解吸离子化飞行时间质谱仪对差异蛋白质点进行了鉴定。结果经肝细胞癌绀织与癌周组织的差异对比分析,共发现有表达水平存在显著差异的35个蛋白质点,14个差异蛋白质点得到了初步鉴定,其中5个蛋白质既往文献中曾报道其与肝细胞癌变相关。结论HBV相关性肝细胞癌蛋白质组与肝硬化组织蛋白质组之间存在差异;本实验研究中初步鉴定的差异蛋白质,可能有助于HBV相关性肝细胞癌的癌变机制、肿瘤标记和治疗靶标的进一步研究。