B7-H1及其受体PD-1在膀胱癌患者外周血免疫细胞中的表达及其临床意义

目的：研究膀胱癌患者外周血中树突状细胞（dendritic cells,DCs）表面B7-H1和CD8＋T淋巴细胞表面PD-1的表达情况及其临床意义。方法：分离30例膀胱癌患者外周血单个核细胞（peripheral blood mononuclear cell,PBMC）,联合培养后采用流式细胞术检测DCs表面B7-HI及CD8＋,T细胞表面PD-1的表达情况。结果：膀胱癌患者外周血DCs表面B7-H1表达水平及CD8＋T细胞表面PD-1表达水平均明显高于正常人,差异具有显著统计学意义（P〈0．01）,并且二者的表达水平随着病理分级和临床分期的增加均升高,差异有统计学意义（P〈0．05）。结论：在膀胱癌发生发展过程中存在着DCs表面B7-H1和CD8＋T细胞表面PD-1分子表达的上调,B7-H1／PD-1信号通路可能在T细胞免疫应答的初始阶段亦参与了膀胱癌免疫逃逸的发生。     

活动性狼疮肾炎患者外周血单个核细胞钙调神经磷酸酶活性的检测及白细胞介素-4表达变化

目的：检测活动性狼疮肾炎（LN）患者外周血单个核细胞（PBMC）白细胞介素-4（IL-4）表达及其与钙调神经磷酸酶（Calcineurin，CaN）活性的关系。方法：体外培养活动性LN患者PBMC，应用发色底物法检测胞浆CaN活性，逆转录PCR检测IL-4mRNA表达。结果：（1）在单纯培养情况下，正常对照组和LN组PBMC均出现一定量CaN活化，活动性LN组显著高于正常对照组[（46．08±5．58）mmol／mg．pr vs（8．81±3．61）mmol／mg．pr，P〈0．01]；在PMA＋Ionomyci刺激下，各组CaN活性均升高，活动性LN组CaN活性明显高于正常对照组[（69．34±12．59）mmol／mg．pr vs（37．12±11．57）mmol／mg．pr，P〈0．01]；（2）LN患者PBMC在单纯培养和PMA＋Ionomycin刺激时IL-4蛋白和mRNA表达均显著高于相应的对照组（P〈0．05）；（3）在单纯培养和PMA＋Ionomycin刺激时，FK506对LNPBMC表达IL-4蛋白和mRNA均有显著抑制作用（P〈0．01）。结论：LN患者PBMC存在CaN过度活化；LN患者PBMC高效表达IL-4与其CaN过度活化密切相关，通过阻断CaN活性可调控IL-4表达。     

IgA肾病患者外周血单个核细胞凋亡率及调控凋亡基因相关指标的检测

目的：探讨IgA肾病（IgAN）活动期与稳定期外周血单个核细胞（PBMC）凋亡率、CD3^＋、CD3^＋CD4^＋、CD3^＋CD8^＋、CD3^＋CD4^＋Fas、CD3^＋CD8^＋Fas、sFas、sFasL等表达的意义。方法：采用ELISA检测sFas、aFasL、IL-1β和流式细胞仪检测PBMC凋亡率等指标。结果：IgA肾病活动期患者PBMC凋亡率、sFas、sFasL明显高于正常人，稳定期sFas、sFasL较正常人有统计学差异（P〈0．01）；CD3^＋CD4^＋Fas、CD3^＋CD8^＋Fas升高（活动期，P〈0．05；稳定期，P〈0．01）；PBMC凋亡率与sFas、sFasL无相关性：sFasL与sFas、CD3^＋CD4^＋Fas、CD3^＋CD8^＋Fas有相关性。结论：IgAN活动期PBMC凋亡率明显高于正常人，导致sFas、sFasL增多，在IgAN中增加的sFas、sFasL可能抑制肾小球系膜细胞凋亡，而且可能是IgAN进展中的一个因素。     

乳腺癌患者B7-CD28共刺激通路的研究

目的探讨乳腺癌患者外周血中CD2 8表达阳性的T细胞数量和患者乳腺癌原代细胞表面B7 1(CD80 )、B7 2 (CD86 )的表达水平 ,以探讨乳腺癌患者共刺激通路是否异常。方法采用流式细胞技术检测乳腺癌患者外周血中CD2 8阳性T细胞与乳腺癌患者原代细胞B7分子的表达 ,并与乳腺良性疾病作比较。结果乳腺癌患者CD2 8阳性的T细胞数明显低于对照组 (t =2 879,P <0 0 5 ) ,特别是CD4/CD2 8双阳性T细胞数更低于对照组 (t=3 0 17,P <0 0 0 1)。乳腺癌患者原代细胞表面B7 1和B7 2表达水平都低于对照组。结论乳腺癌患者外周血中T细胞低表达CD2 8分子 ,乳腺癌细胞低表达B7分子 ,从而影响B7 CD2 8共刺激通路 ,使T细胞不能有效地清除肿瘤细胞。     

地塞米松和川芎嗪对狼疮肾炎外周血单个核细胞白细胞介素—1 …

目的 探讨地塞米松、川芎嗪对活动期,静止期狼疮肾炎（ＬＮ）外周血单个核细胞（ＰＢＭＣ）白细胞介素－１２（ＩＬ－１２）表达的影响。方法 应用地塞米松,川芎嗪对ＬＮ患者ＰＢＭＣ培养,以正常人为对照,分别采用ＥＬＩＳＡ法和半定量ＲＴ－ＰＣＲ法检测细胞上清液及细胞ＩＬ－２和ＩＬ－１２ Ｐ４０ｍＲＮＡ表达。结果ＬＮ活动期,静止期较正常对照组ＩＬ－１２蛋白,基因水平明显增高（Ｐ〈０．０１）。地塞米松明显抑制狼     

自体外周血造血干细胞移植对弥漫性大B细胞淋巴瘤患者PD-1、B7-H1表达及T细胞亚群的影响

目的研究自体外周血造血干细胞移植（auto-PBSCT）对弥漫性大B细胞淋巴瘤（DLBCL）患者程序性死亡因子-1（PD-1）及其配体B7同系物1（B7-H1）表达及T细胞亚群的影响。方法选择浙江大学医学院附属第-医院血液科和温州市中心医院血液科2009年7月至2012年6月接受auto-PBSCT的27例DLBCL患者作为观察组,选择25例同期参加体检的健康人群作为对照组。流式细胞仪检测两组PD-1、B7-H1表达,并分析各组T细胞亚群。结果观察组移植后3个月PD-1阳性表达率低于初次诊断时[（4．6±1．0）％和（12．5±2．1）％],差异具有统计学意义（t=4．47,P〈0．05）。观察组两次检测结果均高于对照组,差异具有统计学意义（t=22．70和25．11,P均〈0．05）。观察组移植后3个月B7-H1阳性表达率低于初次诊断时[（5．7±1．4）％和（16．3±1．7）％],差异具有统计学意义（t=6．78,P〈0．05）。观察组两次B7-H1检测结果均高于对照组（t=25．15和38．06,P均〈0．05）。观察组移植后3个月CD3±T细胞百分比高于初次诊断时[（57±7）％和（44±4）％],差异具有统计学意义（t=6．29,P〈0．05）。观察组两次CD3±T细胞百分比检测结果均低于对照组（t=5．69和7．36,P均〈0．05）。观察组移植后3个月CD4＋T细胞百分比低于初次诊断时[（11±4）％和（29±3）％],差异具有统计学意义（t=5．47,P〈0．05）。观察组两次CD4＋T细胞百分比检测结果均低于对照组,差异具有统计学意义（t=4．61和7．11,P均〈0．05）。观察组移植后3个月CD8±T细胞百分比高于初次诊断时[（32．5±2．7）％和（16．4±3．2）％],差异具有统计学意义（t=9．73,P〈0．05）。与对照组相比,观察组初次诊断后CD8±T细胞百分比较低,而移植后3个月较高（t=10．62和14．71,P均〈0．05）。观察组初次诊断后CD4＋／CD8±比值高于移植后3个月（1．8±0．4和0．4±0．1）,差异具有统计学意义（t=8．57,P〈0．05）。移植后3个月CD4＋／CD8±比值明显低于对照组,差异具有统计学意义（t=11．73,P〈0．05）。结论DLBCL患者存在细胞免疫功能紊乱,auto-PBSCT可以影响DLBCL患者的PD-1、B7-H1表达及T细胞亚群百分比,在-定程度上纠正细胞免疫功能。     

B淋巴细胞活化及凋亡和BlyS、IL-10、IL-6对狼疮性肾炎的影响

目的：探讨B淋巴细胞活化及凋亡、BlyS、IL-10、IL-6对狼疮性肾炎的（LN）影响。方法：以双抗体夹心酶联免疫吸附（ELISA）法测定LN患者外周血中BlyS水平,流式蛋白分析系统（CBA）测定外周血IL-6、IL-10水平,流式细胞仪测定外周血CD19＋CD69＋及CD19＋凋亡率（AV）水平。结果：（1）①活动期和静止期LN患者CD19＋CD69＋、AV水平较正常组显著升高（P〈0.01）;②活动期LN患者IL-10、IL-6、CD19＋CD69＋、AV水平较静止期显著升高（P〈0.01）;③活动期、静止期BlyS水平较正常组差异无统计学意义（P〉0.05）。（2）LN患者活动指数（SLEDAI）与ds-DNA、IL-10、AV之间呈正相关,与BlyS、CD19＋CD69＋水平无相关性;BlyS与ds-DNA、IL-10、CD19＋CD69＋、AV均无相关性;IL-10与ds-DNA、AV呈正相关,与CD19＋CD69＋无相关性。（3）逐步回归分析发现活动指数与C3、凋亡率、CD69、C4有相关性,C4对活动指数影响最大;凋亡率与IL-6、IL-10有相关性,IL-6对凋亡率的影响最大。结论：（1）活动期和静止期LN患者B淋巴细胞活化率及凋亡率均增多;（2）CD19＋CD69＋、CD19＋凋亡率、IL-6、IL-10可反映狼疮的活动性。     

LFA—1共刺激诱导狼疮性肾炎外周血单个核细胞IL—10表达的研究

目的探讨淋巴细胞功能相关抗原-1(LFA-1)对活动性狼疮肾炎(LN)外周血单个核细胞(PBMC)白介素10(IL-10)的调节作用.方法利用半定量RT-PCR技术观察LFA-1共刺激对活动性LN患者PBMC IL-10 mRNA表达的影响.结果单独抗CD3抗体(30 ng/m1)能诱导LN患者PBMC IL 10 mRNA轻微表达(0.43+0.03vs 0.55±0.44,P＜0.05)而在抗CD3抗体(30 ng/m1)存在的情况下,抗LFA-1抗体(2μg/m1,相当于LFA-1配体)共刺激时明显增加了PBMC IL 10 mRNA表达(1.31±0.08vs 0.55+0.04,P＜0.01).LFA-1中和抗体明显抑制了这种共刺激诱导的IL-10 mRNA表达(1.31±0.08vs0.64±0.03,P＜0.01).抗LFA-1抗体单独刺激时并不引起IL-10 mRNA表达变化(P＞0.05).结论LFA-1作为共刺激分子诱导IL-10 mRNA表达可能是其参与LN发病的机制之一.     

CD4＋CD25＋调节性T细胞与Foxp3表达在白癜风发病中的作用

目的：研究CD4＋CD25＋调节性T细胞（CD4＋CD25＋Treg细胞）与Foxp3基因表达与白癜风发病的关系及可能机制。方法：以确诊为进展期白癜风且治疗显效的24例患者为研究对象,患者确诊符合白癜风诊断及疗效标准。以16例健康志愿者作为对照。应用流式细胞术检测正常对照、白癜风患者治疗前后各自外周血中CD4＋CD25＋调节性T细胞的比例。采用密度梯度离心法从治疗前后的外周血中提取单个核细胞,以逆转录聚合酶链扩增方法检测其中Foxp3 mRNA的表达水平。结果：进展期且治疗显效的24例白癜风患者外周血中CD4＋CD25＋调节性T细胞的比例以及CD4＋CD25＋调节性T细胞在总CD4＋T细胞中所占比例明显均低于正常对照组（P〈0．05）。治疗后显效的进展期白癜风患者CD4＋CD25＋调节性T细胞的比例与治疗前相比明显升高（P〈0．05）,CD4＋CD25＋调节性T细胞中Foxp3 mRNA的表达水平比治疗也前明显增加（P〈0．05）。结论：进展期白癜风患者体内存在CD4＋CD25＋调节性T细胞的异常,CD4＋CD25＋Treg细胞的数量的减少和Foxp3表达的降低所造成的CD4＋CD25＋Treg细胞免疫抑制功能爱损可能是白癜风发病的一个因素。     

干扰素-γ和白细胞介素4对狼疮肾炎患者巨噬细胞炎症蛋白-1α产生的效应

目的 观察干扰素（IFN）-γ、白细胞介素（IL）-4对狼疮肾炎（LN）患者外周血单个核细胞（PBMC）巨噬细胞火症蛋白-1α（MIP-1α）产生的效应，以探讨Th1、Th2因子是否对LNMIP-1α产生具有调控作用。方法 Elisa方法检测细胞培养上清液MIP-10633，RT-PCRswimMIP-αmRNA。结果 （1）IFN-γ增强LN患者PBMCMIP-1α蛋白及mRNA产生，（2）IL-4抑制LN患者PBMCMIP-1α蛋白及mRNA产生。（3）IFN-γ、IL-4对正常组MIP-1α产生无显著效应。结论 IFN-γ增强LN患者PBMCMIP-1α产生而IL-4抑制MIP-1α产生。     

大鼠肝脏冷缺血再灌注损伤后B7表达的研究

摘要：目的:探讨B7-1和B7-2在大鼠肝脏冷缺血再灌注损伤时的表达及其免疫学意义。方法:将30只大鼠随机分为3组：A组为假手术组(对照组)；B组为冷缺血20min再灌注24h组；C组为冷缺血30min再灌注24h组。分别取各组之肝脏，采用实时反转录聚合酶链反应(RT-PCR)检测肝组织中B7-1和B7-2mRNA的表达。结果:B7-1mRNA在B，C组表达为0.529±0.089和0.618±0.074，均较A组(0.131±0.012)明显增高(P<0.01)。B7-2mRNA在B，C组表达为0.474±0.132和0.682±0.095，均较A组(0.163±0.054)明显增高(P<0.01)。并且B7-1和B7-2在C组表达明显高于B组(P<0.05)。结论:冷缺血再灌注时肝脏B7-1和B7-2表达上调，增加了肝脏的免疫原性。     

门静脉高压症患者脾切除后外周血B7-1/CD28表达的变化

目的探讨B7-1/CD28对门静脉高压症（portal hypertension,PHT）患者免疫功能的影响和作用。方法采用流式细胞分析技术对30例门静脉高压症患者进行研究,检测门静脉高压症患者行脾切除前后外周血淋巴细胞中以及脾脏中B7-1/CD28的表达情况,分析其与患者淋巴细胞亚群以及免疫功能的关系。对患者外周血B7-1/CD28和淋巴细胞进行检测。利用免疫组化方法检测B7-1/CD28在脾脏中的表达,与10例外伤性脾破裂行脾切除之脾脏组织进行比较。结果门静脉高压症患者外周血中淋巴细胞亚群和B7-1/CD28表达较低,脾切除后短期内升高。其脾脏中表达的B7-1/CD28较正常人群高。结论门静脉高压症患者全脾切除后外周血B7-1/CD28通路被激活,患者免疫功能得到部分恢复。     

HLA-G-treated tolerogenic dendritic cells induce tolerogenic potential by increasing expression of B7-1 (CD80) molecules

BACKGROUND: By consensus, HLA-G has a role in the induction of tolerance via interaction with dendritic cells and manipulation of co-stimulatory molecule expression. The purpose of this study was to demonstrate that HLA-G modified dendritic cells exhibit a decreased ability to stimulate T-cells in vitro due to increased expression of B7-1, which provides regulatory signalling to T-cells as a consequence of binding CD28 and CD152 ligands. METHODS: Bone-marrow cells were cultured from Brown Norway (BN) rat femurs and sorted with anti-rat dendritic cell Ab (OX62) microbeads. Isolated dendritic cells (DCs) were treated with HLA-G tetramer for 3 days. Initially, cells were plated with media, alloantigenic splenocytes, and T-cells and then observed in mixed lymphocyte reaction for thymidine uptake. Also, the cells were analyzed by flow cytometry using antibodies for MHC-II (IA), CD80, and CD86. RESULTS: This study displays that HLA-G-modified DCs decrease induction of an alloproliferative response. In addition, this study demonstrated that HLA-G tetramer treatment decreases CD86 and permits expression of CD80 without altering MHC-II expression. DISCUSSION: This study specifically investigated the role of B7-1 (CD80), showing that HLA-G treatment of immature DCs allows expression of B7-1 (CD80) without altering MHC-II expression and simultaneously decreases B7-2 (CD86) expression. Therefore, a low level of B7-2 (CD86) allows attenuation of T-cell response after activation, both of which are beneficial to immune tolerance. The selective expression of B7-1 (CD80) is important and may serve as a guide for future therapies aimed at transplant tolerance.     

The role of B7 ligands (CD80 and CD86) in CD152-mediated allograft tolerance: a crosscheck hypothesis

BACKGROUND: The regulatory mechanism by which the B7 ligands (CD80 and CD86) direct the CD28/CD152 costimulatory pathways is unclear. This study investigated the role of CD80 and CD86 in a CD152-mediated allograft tolerance model. METHODS: A low-responding cardiac transplant model (BALB/c-->B10.A) with possible long-term acceptance was used. Immunocytochemical and flow cytometric analyses of the graft-infiltrating cells were conducted to characterize this transplant model. The influence of anti-CD80 and anti-CD86 treatments on the proliferation and interleukin (IL)-2 productions of the tolerated splenocytes (SC) was analyzed. The role of CD80 and CD86 in the induction and maintenance of the graft acceptance in this transplant model were also tested. RESULTS: B10.A mice could accept the BALA/c cardiac allografts (11/22), and an anti-CD152 antibody blocked the graft acceptance (10/10). Immunocytochemical and flow cytometric analyses showed that CD152+ cells were predominant among the CD4+ cells infiltrating the 100-day grafts of the B10.A recipients (B10.A-100). Either anti-CD80 or anti-CD86 treatment significantly enhanced polyclonal proliferation and IL-2 production of the B10.A-100 SC. Blockade of either CD80 or CD86 prohibited the tolerance transmitted by adoptive transfer, and anti-CD80 or anti-CD86 plus skin grafting undermined the established allograft tolerance. CONCLUSIONS: Both CD80 and CD86 were essential for the induction and maintenance of the CD152-mediated allograft tolerance.     

共刺激分子B7-H3 mRNA和B7-H3蛋白在胃癌组织中的表达及其临床意义

目的研究B7家族的新成员B7-H3mRNA和B7-H3蛋白在胃癌组织中的表达及其临床意义。方法采用荧光实时定量逆转录-多聚酶链反应（RT—PCR）方法检测38例胃癌患者癌组织及其邻近正常组织B7-H3mRNA的表达；免疫组织化学Elivision法检测B7．H3蛋白在胃癌组织中的表达。结果B7-H3mRNA在胃癌组织及其邻近正常组织中均有表达，其在胃癌组织中的表达明显低于邻近正常组织。B7-H3mRNA的表达与胃癌患者性别、年龄、组织类型、肿瘤大小、淋巴结转移、病理分级和浸润深度无关（P〉0.05）。B7-H3蛋白在胃癌组织中表达的阳性率为39、5％，其与患者性别、年龄、组织类型、肿瘤大小、淋巴结转移和浸润深度也无关（P〉0.05）。B7-H3蛋白表达随肿瘤临床分期的递增而降低，各组间差异有统计学意义（P=0.022）；病理分级各组间差异也有统计学意义（P=0.039）。Kaplan—Meier生存分析显示，B7-H3阳性表达组无病生存期明显优于B7-H3阴性表达组（P=0.009），B7-H3阳性表达组总生存期亦明显优于B7-H3阴性表达组（P=0.010）。结论B7-H3表达与胃癌预后有关，可以作为反映患者免疫状态和预后的一个新的指标．为制定临床治疗方案提供依据。     

CD1D基因与B7-1基因共转染胰腺癌细胞及其抗癌作用的实验研究

目的:观察共转染小鼠B7-1和CD1D 基因对小鼠胰腺癌细胞的免疫应答。方法:将表达B7-1和CD1D 基因的逆转录病毒载体导入包装细胞系,然后转染入胰腺癌细胞。用PCR和Western方法鉴定细胞表达;用B7-1和CD1D 阳性的细胞在体内诱导抗肿瘤免疫反应。结果:B7-1和CD1D 阳性的细胞在同基因小鼠体内能诱导抗肿瘤免疫反应并抑制肿瘤生长。结论:B7-1基因和CD1D 基因共转染肿瘤细胞,能抑制肿瘤生长,可能为肿瘤的基因治疗开辟一条新途径。     

Usefulness of inhibiting the lymph node metastasis in human gastric carcinoma by B7-1 gene transfection

INTRODUCTION: Lymph node metastasis is one of the crucial prognostic factors in gastric cancer. We have reported that ICAM-1 gene transfection was effective against lymph node metastases of gastric cancer. B7-1, one of the co-stimulatory factors, was reported to induce cytotoxic T lymphocytes when using melanoma and bladder cancer cell lines, as well as ICAM-1. In this study, we investigated the inhibitory effect of B7-1 on lymph node metastasis by B7-1 gene transfection into gastric cancer cells. MATERIALS AND METHODS: We transfected B7-1 genes into a gastric cancer cell line (OCUM-2MLN) and analyzed the effect of B7-1 transduction on lymph node metastasis, the in vitro adhesiveness and cytotoxicity assay of mononuclear lymphocytes to cancer cells and lymph node metastatic ability after orthotopic implantation of gastric cancer cells in vivo. RESULTS: We revealed that mononuclear lymphocytes showed significantly stronger adherence and cytotoxicity to B7-1 transfected cells (2MLN/B7) than its parent OCUM-2MLN cells. The tumor growth rate of 2MLN/B7 xenograft was significantly slower than OCUM-2MLN xenograft in nude mice. In orthotopic implantation experiments for nude mice, 2MLN/B7 cells in stomach developed significantly less lymph node metastasis than OCUM-2MLN cells. Histologic findings showed that leukocytes were intensively infiltrated in both the 2MLN/B7 tumors and its metastatic lesions, however, were scarcely observed in the lesions associated with 2MLN cells. CONCLUSION: B7-1 may play an important role in inhibiting lymph node metastasis by the mechanism of enhanced immunogenicity, and that B7-1 gene transduction might be effective against lymph node metastases of gastric cancer.     

实时荧光定量聚合酶链反应检测B7-H4基因表达的方法学构建

目的 建立实时荧光定量聚合酶链反应(PCR)检测协同刺激分子B7-H4基因表达的方法.方法 采用自行设计的检测协同刺激分子B7-H4及内参照基因GAPDH的引物及TaqMan探针,经优化反应体系及反应条件后,检测B7-H4及GAPDH的基因表达.将B7-H4和内参照基因GAPDH的PCR扩增产物经测序鉴定正确并纯化后分别与pMD19-T载体连接,克隆构建含B7-H4及GAPDH基因片段的重组质粒,作为实时荧光定量PCR检测B7-H4及GAPDH基因表达的标准品.将重组质粒标准品进行定量及梯度稀释,建立标准曲线.结果 PCR扩增产物分别经测序证实为B7-H4及GAPDH基因的特异性片段.该法检测B7-H4基因表达的灵敏度为527 copies/ml,线性范围为5.27×102～5.27×107 copies/ml,标准曲线方程为:Y=-3.1395X+41.805,直线回归相关系数r=0.995,批间变异系数范围为2.39%～3.59%,扩增效率108.2%.该法检测GAPDH基因表达的灵敏度为386 copies/ml,线性范围为3.86×102～3.86×107copies/ml,标准曲线方程为:Y=-3.2436X+41.083,直线回归相关系数r=0.999,批间变异系数范围为2.26%～3.86%,扩增效率103.4%.结论　成功构建实时荧光定量PCR检测协同刺激分子B7-H4基因表达的方法,该方法 特异性好、灵敏度高、重复性好.

Abstract：

Objective To establish a real-time polymerase chain reaction (PCR) method to detect costimulatory molecule B7-H4 gene expression. Methods The mRNA levels of costimulatory molecule B7-H4 and the reference gene GAPDH were detected by using real-time PCR using TaqMan technology. The primers and TaqMan probes of B7-H4 and GAPDH were designed. The B7-H4 and internal reference gene GAPDH fragments in pure form from classical PCR amplification were cloned into pMD19-T vector, and recombinant plasmids were used as the standard substances of B7-H4 and GAPDH quantitative detection.Standard curves were established using a serial dilution of quantified plasmids to measure B7-H4 and GAPDH. The reaction systems were optimized, and the sensitivity, specificity and reproducibility were evaluated. Results The amplification products were confirmed as the specific fragments of B7-H4 and GAPDH by DNA sequencing instrument. For B7-H4, the sensitivity was 527 copies/ml. The linear range was from 5.27 × 102 to 5.27 × 107 copies/ml, the standard curve equation was Y = - 3. 1395X +41. 805, the correlation coeffecient was 0. 995, the interassay coefficient of variations was 2. 39% -3.59%, and the amplification efficiency was 108.2%; For GAPDH, the sensitivity was 386 copies/ml. The linear range was from 3. 86 × 102 to 3.86 × 107 copies/ml, the standard curve equation was Y = - 3. 2436X +41. 083, the correlation coeffecient was 0. 999, the interassay coefficient of variations was 2. 26% -3. 86%, and the amplification efficiency was 103.4%. Conclusion The real-time PCR system for quantifying costimulatory molecule B7-H4 mRNA levels has been established successfully with high specificity, sensitivity and good repeatability.     

胰腺癌中磷酸酶和张力蛋白同源物基因及B7-H1蛋白的表达及其临床意义

目的 观察胰腺癌磷酸酶和张力蛋白同源物基因(PTEN)蛋白和B7-H1蛋白的表达,探讨两者与胰腺癌肿瘤生物学特征的关系.方法 采用免疫组织化学ELPS法检测43例胰腺癌患者的组织标本和作为对照的5例非癌胰腺标本中PTEN和B7-H1蛋白的表达水平,分析其与临床和病理学特征的关系.结果 比较非癌胰腺组织,胰腺癌组织中PTEN蛋白表达显著降低,而B7-H1蛋白表达显著升高(P＜0.01),两者的表达还呈现显著负相关(r=-0.414,P＜0.01);两者的表达水平与肿瘤的分化程度、TNM分期、浆膜侵犯及淋巴结转移有关(P＜0.05),B7-H1表达水平还与年龄显著相关(P＜0.01);两者分别的表达水平与患者的预后均显著相关(P＜0.01),两者的联合表达水平与患者的预后相关(P＜0.05).结论 mN和B7-H1蛋白的表达水平与胰腺癌的发生、发展和预后密切相关.

Abstract：

Objective To investigate the probable correlation between the expressions of phosphatase and tensin homologue deleted on chromosometen (PTEN) and B7-H1 protein in pancreatic carcinoma and the biological behavior characteristics of tumors. Methods Forty-three patients were recruited who had undergone surgical resection for pancreatic carcinoma between 2002 and 2009. The PTEN and B7-H1 protein expressions in the tissue specimens of these 43 patients and 5 non-pancreatic carcinoma people' s pancreatic tissue specimens were evaluated by immunohistochemistry ELPS technique, and the clinical and pathological features of these specimens and the follow-up information were analyzed. Results PTEN expressions were significantly lower in pancreatic carcinoma tissues than in non-pancreatic carcinoma people' s pancreatic tissues but B7-H1 expressions were significantly higher ( P ＜ 0. 01 ). The expression of PTEN was negatively correlated to that of B7-H1 (r = -0.414 ,P ＜0. 01). PTEN and B7-H1 expressions correlated with the pathological grade and tumor-node-metastasis ( TNM ) stage, peripancreatic invasion, regional lymph node involvement,respectively (P＜0. 05). B7-H1 expressions also significantly correlated with the ages (P＜0. 01). Furthermore, PTEN and B7-H1 expressions showed significant prognostic effects (P＜0.01) and there are correlations existed between combined PTEN/B7-H1 expression and prognostic effects (P ＜0. 05). Conclusion The expression of PTEN and B7-H1 may be significantly correlated to the carcinogenesis,development and prognosis of pancreatic carcinoma.