慢性乙肝患者外周血CD4CD25FOXP3 Tregs及HBV抗原特异性CTLs的检测和分析

目的: 检测慢性乙肝(CHB)患者外周血中CD4⁺CD25⁺FOXP3⁺调节性T淋巴细胞(Treg细胞)和乙肝病毒(HBV)特异性细胞毒性T淋巴细胞(CTLs)的表达及意义。方法: 收集28例CHB患者和15例健康人外周血单个核细胞标本,运用流式细胞仪对Treg细胞亚群进行定量分析,同时采用酶联免疫斑点法检测HBV抗原特异性CTLs,并结合丙氨酸氨基转移酶(ALT)和 HBV DNA的临床情况进行分析。结果: CHB组CD4⁺CD25⁺FOXP3⁺ Treg细胞的频率显著高于健康对照组 (3.14%±0.97% vs 1.95%±0.68%,P＜0.05);HBV抗原特异性CTL斑点计数为阳性(19.28±3.85)。CHB组Treg的频率与乙肝病毒载量呈正相关(r=0.831, P＜0.01),与HBV特异性CTL斑点计数值呈负相关(r＝-0.540,P＜0.01)。结论: CHB患者外周血CD4⁺CD25⁺FOXP3^＋Treg细胞表达升高并与病毒载量相关,而与HBV反应的CTLs数量呈负相关,提示Treg细胞可通过抑制细胞免疫反应影响病毒清除。     

Th17细胞联合CD3+CD8-IL-21+T细胞升高与宫颈癌发生发展的关系

目的: Th17细胞在免疫调节中起重要作用,而IL-21与Th17在分化调节和功能行使上密切相关。本研究旨在探讨Th17在宫颈癌发生发展中的作用。方法: 选取37例宫颈癌患者、25例宫颈上皮内瘤变(CIN)患者和18例健康志愿者作为研究对象,用流式细胞分析术检测外周血中Th17细胞及CD3⁺CD8^-IL-21⁺T细胞的比例。分析两者与临床病理指标之间的关系。结果: 与健康对照组相比,Th17细胞及CD3⁺CD8^-IL-21⁺ T细胞比例(占淋巴细胞百分比)在CIN组(P<0.01,P<0.05)及宫颈癌组(P<0.01,P<0.05)均明显升高。此外,2种细胞的比例都与临床分期有关,在晚期宫颈癌组明显升高(均P<0.05),并且有淋巴结转移组或脉管浸润组都明显高于相对应的无转移组(P<0.01, P<0.05)或无浸润组(均P<0.01)。此外,在健康对照组和宫颈癌组,Th17与CD3⁺CD8^-IL-21⁺ T细胞呈正相关,CD3⁺CD8^-IL-21⁺ T细胞的比例还与肿瘤大小有关(P<0.01)。结论: Th17和CD3⁺CD8^-IL-21⁺ T细胞在宫颈癌患者外周血中的比例上调,在宫颈癌的发生发展中可能起着重要作用。     

系统性红斑狼疮患者外周血淋巴细胞表达BLyS和CD38的变化

目的:研究系统性红斑狼疮(SLE)患者外周血淋巴细胞表达BLyS和CD38的变化。方法:收集22名SLE患者和14名健康人外周血淋巴细胞, 用流式细胞仪检测外周血淋巴细胞表达BLyS和CD38的变化。结果:SLE患者外周血BLyS⁺淋巴细胞、CD19⁺淋巴细胞和CD19⁺CD38⁺淋巴细胞显著增加, BLyS⁺淋巴细胞增加与CD19⁺CD38⁺淋巴细胞增加呈正相关(r=0.434, P<0.05).结论:SLE患者外周血淋巴细胞表达BLyS和CD19⁺B淋巴细胞表达CD38均显著增加, 且二者增加呈正相关。     

Foxp3转染对哮喘小鼠脾淋巴细胞功能的影响

目的: 转染Foxp3至哮喘小鼠脾淋巴细胞,探讨Foxp3表达对脾淋巴细胞功能的影响。方法: 卵白蛋白(OVA)致敏激发制作哮喘小鼠模型,收集培养脾脏淋巴细胞;使用电穿孔法转染真核表达载体pcDNA3.1(-)-Foxp3至脾脏淋巴细胞,并设转染空质粒组和对照组;RT-PCR和Western blotting检测Foxp3的表达;流式细胞术检测转染后CD4⁺CD25⁺ Treg细胞/CD4⁺细胞比例;MTT法检测转染后的脾脏淋巴细胞增殖反应,ELISA检测脾淋巴细胞上清中白细胞介素4(IL-4)和干扰素γ(IFN-γ)的含量。结果: 转染组Foxp3 mRNA 和蛋白的表达水平显著高于空质粒组和对照组;转染Foxp3后CD4⁺CD25⁺ Treg细胞/CD4⁺细胞比例显著高于空质粒组和对照组;与空质粒组和对照组相比,转染pcDNA3.1(-)-Foxp3质粒明显抑制了脾淋巴细胞增殖;转染组细胞上清中IL-4和IFN-γ含量低于空质粒组和对照组。结论: 转染pcDNA3.1(-)-Foxp3至哮喘小鼠脾淋巴细胞,Foxp3得到有效表达。Foxp3的高表达能增加CD4⁺CD25⁺ T细胞的数量,抑制哮喘小鼠脾淋巴细胞的增殖以及Th1和Th2细胞因子的产生。     

自然流产中外周血和蜕膜组织自然杀伤细胞亚群的比例变化

目的和方法:采用流式细胞仪检测淋巴细胞亚群法探讨自然流产与正常早孕之间外周血和蜕膜自然杀伤细胞亚群的差异。结果:外周血中自然流产组的CD56⁺的百分率较早孕组有减少的趋势,而CD56⁺CD16⁺的百分率则较早孕组显著减少,CD16⁺的百分率两组间无差异。自然流产组的蜕膜CD56⁺、CD56⁺CD16⁺、CD16⁺的百分率均明显低于早孕组。结论:蜕膜中CD56⁺NK细胞的减少可能是自然流产的原因之一,外周血中CD56⁺和CD56⁺CD16⁺NK细胞的丢失可能对自然流产的发生具有诊断价值。     

镁对哮喘患者外周血CD4+ CD25+调节性T细胞凋亡及Foxp3表达的影响

目的:观察镁对分离培养的健康人和哮喘患者外周血CD4⁺CD25⁺调节性T细胞凋亡及叉头框蛋白3(Foxp3)表达的影响。方法:经磁珠分离法分离出健康人和哮喘患者外周血CD4⁺CD25⁺T细胞,分镁剂干预组(10 mmol/L)及空白组培养72 h后,用流式细胞仪检测CD4⁺CD25⁺T细胞的凋亡率及Foxp3表达情况。结果:(1)健康人外周血CD4⁺CD25⁺T细胞的纯度为77.4％~92.3%,哮喘患者CD4⁺CD25⁺T细胞的纯度为75.2％~93.8％。(2)CD4⁺CD25⁺T细胞占外周血CD4⁺T细胞的比例在健康组为4.12%~7.98%,在哮喘组为4.51%~8.68%,两者没有显著差异(P＞0.05)。(3)镁(10mmol/L)可以诱导健康组及哮喘组外周血CD4⁺CD25⁺T细胞凋亡率增加(P＜0.05),但对Foxp3的表达无影响(P＞0.05)。结论:镁促进CD4⁺CD25⁺T调节细胞凋亡增加可能为其治疗支气管哮喘的作用机制之一。     

CIK细胞回输联合吉西他滨和顺铂治疗鼻咽癌放疗后肝肺转移瘤的效果及机制研究

目的: 观察评价细胞因子诱导杀伤(CIK)细胞回输联合吉西他滨和顺铂(GP)方案化疗治疗鼻咽癌放疗后肝肺转移瘤的近远期疗效,并探讨其机制。方法: 2007年8月至2008年7月放疗后随访发现肝或肺转移患者30例,随机分为3组。研究1组10例行过继性CIK细胞回输联合GP化疗;研究2组10例行单纯GP化疗。对照组患者及健康志愿者各10例,不给予抗肿瘤治疗而仅进行随访。观察其近远期疗效、血清EB病毒DNA PCR定量和外周血淋巴细胞亚群分布变化。结果: CIK+GP组有效率(90%)比单纯GP组(70%)好,但2组间无明显差异;而CIK+GP组和GP组均比对照组(10%)好,差异明显。CIK+GP组最和GP组治疗后血清EBV-DNA PCR定量均有不同程度的下降,尤以CIK+GP组为明显,而对照组则无明显改变。鼻咽癌放疗后肝肺转移瘤患者外周血CD3⁺比例较健康志愿者显著低下,经CIK+GP治疗后CD3⁺比例较单纯GP化疗组有所提高;肝肺转移瘤患者和健康志愿者CD4⁺/CD8⁺比例无明显差异,经CIK+GP生物化疗后其比例明显升高,而经GP化疗后明显降低,2组之间存在明显差异。CIK+GP组、GP组和对照组2年生存率(OS)分别为60.0%、40.0%和20.0%,生存曲线分析显示3组病例之间均有明显差异(P＜0.05)。结论: CIK 细胞回输联合GP 化疗治疗鼻咽癌放疗后肝肺转移瘤具有肯定的近远期疗效并可改善其预后;两者具有协同作用,其作用可能与改变CD3⁺及CD4⁺/CD8⁺比例有关。     

不同宫颈癌临床分期患者外周血中T淋巴细胞及NK细胞的表达分析

目的分析宫颈癌患者外周血中T淋巴细胞及其亚群以及NK细胞的表达情况。方法前瞻性分析145例在我院就诊的宫颈癌患者的临床资料,按照宫颈癌FIGO临床分期差别分为:宫颈癌Ⅰ期38例、宫颈癌Ⅱ期42例、宫颈癌Ⅲ期35例、宫颈癌Ⅳ期30例;同期选择体检正常女性30例作为对照组。应用流式细胞仪检测各组患者外周血CD3^+、CD4^+、CD8^+T细胞亚群、调节性T细胞(Treg)以及NK细胞数量,同时计算各亚群的比例以及Treg占CD4^+T细胞的比例。应用多元Logistic回归分析评估各细胞亚群数量及比例与宫颈癌临床分期的相关性。结果与对照组相比,宫颈癌患者CD3^+T细胞、CD4^+T细胞、NK细胞数量以及CD4^+/CD8^+比值均较低,Treg/CD4^+比值较高(均P <0.05),而CD8^+T细胞数量差异无统计学意义(均P>0. 05)。随宫颈癌病理严重程度递增,CD3^+T细胞、CD4^+T细胞、NK细胞数量以及CD4^+/CD8^+比值逐步降低,Treg/CD4^+比值逐步增高(均P <0.05),而CD8^+T细胞数量差异无统计学意义(均P>0.05)。多元Logistic回归分析显示,Treg/CD4^+比值是宫颈癌临床分期的危险因素,CD4^+T细胞、CD4^+/CD8^+比值及NK细胞是其保护性因素(均P<0.05)。结论宫颈癌患者细胞免疫功能均不同程度降低,晚期患者降低最显著。检测T淋巴细胞亚群及NK细胞可用于宫颈癌患者免疫监测,为临床治疗及预后评估提供参考。     

Treg/Th17失衡在脓毒症发病机制中的作用

目的: 观察脓毒症患者外周血中CD4⁺CD2⁺CD127^-调节性T细胞(Treg)及辅助性T细胞17(Th17)的比例,并探讨Treg/Th17失衡在脓毒症发病中的作用。方法: 选择2010年1~8月入住我院ICU的脓毒症患者70例,按疾病严重程度分为脓毒症组(34例)、重症脓毒症组(21例)和脓毒症休克组(15例)。第1 d抽取外周血,应用流式细胞术检测不同组别患者外周血中Treg/Th17比例及CD14单核细胞 HLA-DR表达率,并探讨Treg/Th17比例与免疫状态及病情严重程度(APACHEⅡ评分)的关系。选取同期健康查体者为对照组(30例)。结果: (1) 和对照组相比,各病例组外周血Treg及Th17比例均明显升高(均P<0.01),但升高的程度不同。Th17表达以重症脓毒症组升高最明显,Treg表达以脓毒症休克组升高最明显。(2)Treg/Th17:脓毒症休克组>重症脓毒症组>脓毒症组,各组两两相比均有显著差异(均P<0.01)。脓毒症组Treg/Th17显著低于对照组(P<0.01),脓毒症休克组Treg/Th17显著高于对照组(P<0.01),但重症脓毒症组Treg/Th17与对照组比较无明显差异。(3)CD14单核细胞HLA-DR表达率:脓毒症休克组<重症脓毒症组<脓毒症组<对照组,两两比较均有显著差异(均P<0.01)。(4)Th17比例和APACHEⅡ评分及 HLA-DR表达率无相关性;Treg表达率和APACHEⅡ评分正相关(r=0.93, P<0.01),和HLA-DR表达率负相关(r=-0.89, P<0.01);HLA-DR表达率和APACHEⅡ评分负相关(r=-0.91, P<0.01)。结论: (1)脓毒症患者中Treg和Th17比例失调,Treg/Th17的失衡参与脓毒症的发病及进展。(1)Treg表达率可以在一定程度上反映机体病情及免疫状态。     

类风湿关节炎CD4+CD28-T细胞与淋巴细胞凋亡的相关性研究

目的: 研究类风湿关节炎(RA)患者外周血CD4⁺CD28^-T细胞比例与淋巴细胞凋亡异常的相关性。方法: 采用流式细胞术三色分析法检测50例患者和50例健康志愿者的外周血淋巴细胞中CD4⁺CD28^-T细胞比例;通过加入PHA孵育检测RA病人外周淋巴细胞和正常对照的淋巴细胞对激活诱导细胞死亡(AICD)易感性差异;分析CD4⁺CD28^-T细胞比例与外周血淋巴细胞凋亡率的相关性。结果: RA组CD4⁺CD28^-T细胞比例的均数明显高于健康对照组(7.79%±3.52% vs 1.89%±1.78%,P<0.05)。RA组病人外周血淋巴细胞的AICD凋亡率低于健康对照组(11.38%±5.73% vs 19.46%±6.32%,P<0.05)。Spearman相关分析结果显示CD4⁺CD28^-T细胞比例与外周血淋巴细胞AICD凋亡率负相关(r=-0.433,P<0.01)。结论: RA患者外周血中CD4⁺CD28^-T细胞比例增多,活化淋巴细胞生存期延长,这可能参与RA的发病机制。     

AFP-L3在肝细胞癌早期诊断和疗效监测中的价值

为了探讨甲胎蛋白异质体(AFP-L3)在肝细胞癌的早期诊断和疗效监测价值,本研究联合应用微量离心柱法和电化学发光法随访监测83例原发性肝细胞癌患者和57例首诊为非肿瘤性慢性肝病患者血清中的AFP和AFP-L3水平。原发性肝细胞癌患者监测时间为肝癌术前至术后6个月,慢性肝病患者监测时间为首诊前至诊后2年。同时收集原发性肝细胞癌患者门脉癌栓情况、肿瘤分化程度、临床分期、肿块大小等临床诊断指标。研究结果为,随访的22例AFP-L3持续阳性的慢性肝病患者平均3.15个月均确诊为肝癌,肝癌发生率为45.45%;4例AFP-L3阴性变阳性的慢性肝病患者,平均8.17个月均确诊为肝癌,肝癌发生率为100%;31例AFP-L3持续阴性的慢性肝病患者,平均11.75个月确诊为肝癌,肝癌发生率为9.68%。各组相比差异有统计学意义。原发性肝细胞癌患者手术有效者AFP和AFP-L3水平明显下降,而病情稳定和进展者二者水平无显著变化或升高。AFP-L3水平与肿瘤分化程度相关,而与门脉瘤栓、临床分期和肿块大小不相关。在有门脉瘤栓组、低分化肝癌组、Ⅰ期肝癌组和小肝癌组中,其AFP-L3的阳性率分别为81.97%、100%、75%和77.78%。由此可知AFP-L3在肝细胞癌早期诊断和疗效监测中具有重要的临床价值。     

ELISA法检测甲胎蛋白异质体用于肝癌诊断的探讨

目的应用ELISA检测甲胎蛋白异质体(AFP-L3),并探讨AFP-L3浓度在HCC组与良性肝病组诊断与鉴别诊断的价值。方法用ELISA法检测137例AFP阳性的肝病患者血清AFP-L3浓度,用ROC曲线分析AFP-L3。结果 92例HCC患者AFP-L3浓度为109.04±62.51ng/mL,明显高于45例良性肝病组(25.96±49.43ng/mL)P0.001。HCC的ROC曲线面积为0.819,以AFP-L3浓度37.89ng/mL为临界值,分析HCC患者与良性肝病患者AFP-L3浓度异常的敏感性为83.7%,特异性为88.9%,诊断正确率为85.4%。结论用ELISA法检测AFP-L3浓度对HCC诊断与良性肝病鉴别诊断有较高的临床价值,操作简便,费用低廉。     

甲胎蛋白异质体L3预警原发性肝癌的研究

目的 探讨甲胎蛋白异质体(AFP-L3)的检测在预警原发性肝癌中的作用.方法 对100例AFP升高肝病患者血清,应用甲胎异质体微量离心柱分离并洗脱获得AFP-L3,再同时检测原始血清中的AFP以及AFP-L3含量,计算AFP-L3在AFP中的比例,对AFP-L3异常升高者、正常者进行跟踪,结合6个月后临床诊断结果 ,分析AFP-L3升高在鉴别良性肝脏病变与预警肝癌中的作用.结果 肝癌、疑似肝癌患者与良性肝病患者中的AFP-L3阳性率差异有统计学意义(分别为81.80%、73.68%、11.80%,P＜0.05).未确诊肝癌(疑似HCC、肝病)的患者中,AFP-L3阳性的21例中有12例在6个月内被诊断为HCC,而且有6例是通过B超、CT等影像学手段被早期确诊的单发性小肝癌.AFP-L3阴性的62例标本中,6个月内有3例发生肝癌,AFP-L3阳性发生肝癌的危险率增加了11.9倍.结论 AFP-L3与AFP值无相关性,可以作为一个独立肝细胞癌诊断因子.AFP-L3的测定对于AFP升高时良、恶性肝病的鉴别及肝癌的早期预警诊断具有重要意义.     

甲胎蛋白异质体L3对低浓度甲胎蛋白肝癌诊断的临床意义

目的探讨在低浓度甲胎蛋白（AFP）肝病患者中,甲胎蛋白异质体L3（AFP-L3）的百分含量（AFP—L3％）对肝癌的早期诊断和疗效评估的临床意义。方法收集245例血清低AFP含量（5～40ng／m1）的肝病患者样本（其中肝硬化患者100例、肝癌患者145例）,检测AFP—L3％,并对其中100例肝硬化患者和20例肝癌术后患者分别进行3个月和12个月的随访。结果以AFP—L3％≥10％作为阳性判断标准,100例肝硬化患者中阳性为23例,经3个月随访后其中8例诊断为肝癌;145份肝癌患者血清AFP-L3％阳性率为46．2％（67／145）。低浓度AFP肝癌组的AFP—L3％水平显著高于低AFP肝硬化组（t=7．318,P=0．001〈0．01）;20例肝癌患者术后有5例AFP．L3％仍为阳性,其在12个月内的生存率为0,而术后AFP—L3％阴性患者生存概率为15／15。结论AFP—L3％在低浓度AFP肝病患者中对肝癌的早期诊断和术后疗效评估都具有一定的临床意义。     

Evaluation of tumor markers for the detection of hepatocellular carcinoma in Yangon General Hospital,Myanmar

Levels of alpha-fetoprotein (AFP), its glycoforms AFP-L3 and AFP-P4, and proteins induced by vitamin K absence or antagonist-II (PIVKA-II) were determined in sera obtained from patients in Yangon General Hospital (20 with hepatocellular carcinoma (HCC), 29 with chronic liver diseases, including 3 with chronic hepatitis and 26 with cirrhosis of the liver, and 9 with other hepatobiliary diseases). Forty-five percent of the patients with HCC had serum AFP levels above 10,000 ng/ml, indicating that nearly half of the HCC patients were at an advanced stage of the disease. Thus, the AFP sensitivity was as high as 70% with 100% specificity for a cutoff level of 200 ng/ml. The sensitivity of AFP-L3 was 75% and a specificity 90% for a cutoff level of 15%. AFP-P4 showed a higher sensitivity of 80% and a similar specificity of 86% for a cutoff level of 12%. Combined evaluation of AFP-L3 and/or AFP-P4 increased the sensitivity to 90% with the same specificity of 86%, indicating that AFP-L3 and AFP-P4 are useful as adjuncts for diagnosis of HCC in the present population. PIVKA-II had a high sensitivity of 90%, although the specificity was lower than 45%, probably due to the low cutoff level, as some cholestatic patients were included in the control group.     

微量离心柱法检测AFP-L3的临床应用研究

目的 探讨微量离心柱法检测甲胎蛋白异质体在肝癌预警及良恶性肝病鉴别诊断中的临床价值.方法 应用装有耦联小扁豆凝集素（LCA）的微量离心柱分离300例肝病患者的AFP-L3,采用化学发光法检测AFP及AFP-L3,计算AFP-L3占总AFP的比率（判断标准以AFP-L3≥10%者为阳性）.结果 AFP-L3在肝细胞癌患者组中的阳性率是95%,在慢性肝病患者组的阳性率是64%,两组患者AFP-L3阳性率差异有统计学意义（x2=134.72,P＜0.01）;AFP-L3阳性的慢性肝病患者与阴性患者肝癌发生率差异有统计学意义（x2=80.158,P＜0.01）;AFP-L3的百分比与AFP浓度不相关（r=0.046,P＞0.05）.结论 微量离心柱法检测甲胎蛋白异质体（AFP-L3）在肝细胞癌诊断、预警及与良恶性肝病的鉴别诊断中具有重要价值.     

Usefulness of the Novel Oncofetal Antigen Glypican-3 for Diagnosis of Hepatocellular Carcinoma and Melanoma

Glypican-3 (GPC3) mRNA and protein are expressed in >80% of human hepatocellular carcinomas (HCC) but not in normal tissues except for placenta and fetal liver. The oncofetal antigen GPC3 is a glycosylphosphatidyl inositol-anchored membrane protein and may be secreted. It is a novel tumor marker for human HCC: GPC3 protein was present in sera from 40-50% of HCC patients, but was not detected in sera from patients with liver cirrhosis or chronic hepatitis, or in sera from healthy individuals. alpha-Fetoprotein (AFP) and PIVKA-II (protein induced by vitamin K absence or antagonist-II), are well known major tumor markers for HCC. Generally, AFP shows high positivity for HCC but also high false-positivity in detection assays. Lens culinaris agglutinin-reactive fraction of alpha-fetoprotein (AFP-L3) is a recently described marker of HCC. Detection of AFP-L3 shows a much higher specificity than AFP, but a lower sensitivity. On the other hand, detection of PIVKA-II shows a lower false-positivity, but is not always sensitive enough to detect low levels secreted by small HCCs. There was no correlation between the three tumor markers, AFP, PIVKA-II, and GPC3 in terms of their presence in HCC cells. All three tumor markers showed similar positivity in patients with HCC, detecting 80% of patients with the disease. GPC3 is also a novel tumor marker for the diagnosis of human melanoma, especially in the early stages of the disease. Expression of GPC3 mRNA and protein was evident in tumor cells from >80% of patients with melanoma and melanocytic nevus, which is a common benign lesion. GPC3 protein was detected in sera from 40% (36/91) of melanoma patients, but not in sera from those with large congenital melanocytic nevus, or from healthy donors. Surprisingly, we detected serum GPC3 even in patients with stage 0, in situ melanoma. The positive detection rate of serum GPC3 at stage 0, I, and II (44.4%, 40.0%, 47.6%, respectively) was significantly higher than that of 5-S-cysteinyldopa, a well known tumor marker for melanoma (0.0%, 8.0%, and 10.0%, respectively). Interestingly, GPC3 was highly immunogenic in mice and elicited effective anti-tumor immunity with no evidence of autoimmunity. Thus, GPC3 is useful for diagnosis of HCC and melanoma and may also have a role in immunotherapy or tumor prevention. However, studies in humans are warranted.     

Additive effects of amyloid beta fragment and interleukin-1beta on interleukin-6 secretion in rat primary glial cultures

Serum concentration levels of des-gamma-carboxy prothrombin (DCP), alpha-fetoprotein (AFP) and Lens culinaris agglutinin-reactive fraction (AFP-L3) are useful tumor markers for the diagnosis of hepatocellular carcinoma (HCC). Recently, a novel immunoassay using the electrochemiluminescence (ECLIA) was developed to enable measurement of low-concentration of DCP. This study investigated the usefulness of high-sensitive DCP for the early diagnosis of HCC. The subjects consisted of 90 patients with viral cirrhosis who could be followed for at least 24 months from 1992 to 1997. Fifty-six of these patients developed HCC and 34 patients had not by 1998. We measured the serum levels of high-sensitive DCP, AFP and %AFP-L3 every 3 months during 2 years before the detection of tumor in patients with HCC and during 2 years from 1995 to 97 in patients without HCC. Youden's index was calculated for evaluation of the ideal cut-off levels. The patterns of serial changes during 2 years were divided into two types: fluctuating type and non-fluctuating type. Cut-off levels of 40 mAU/ml for high-sensitive DCP (Youden's index = 0.435), 20 ng/ml for AFP (Youden's index = 0.442) and 10% for %AFP-L3 (Youden's index = 0.364) gave the highest index for each marker. When these markers were combined, the combination of high-sensitive DCP, AFP and %AFP-L3 gave the highest accuracy (sensitivity = 82.1%, specificity = 82.4%, accuracy = 82.2%). Fluctuating type of high-sensitive DCP, AFP and %AFP-L3 levels were found in 15 (17%), 29 (32%) and 11 (12%) patients, respectively. The rate of complication with HCC in the patients who showed the fluctuating type of high-sensitive DCP levels was significantly greater than that in the patients who showed non-fluctuating type (P<0.01). These results suggest that periodic measurement of serum DCP levels using ECLIA method is very useful for HCC screening and predicting the development of HCC.     

血清中甲胎蛋白异质体L3含量与肝癌分期及大小的相关性

目的 探讨原发性肝癌患者血清中的甲胎蛋白异质体L3（AFP-L3）水平与肝癌分期和癌灶大小的相关性.方法 随机选取162例原发性肝癌（PHC）患者的血清,分别进行总甲胎蛋白（AFP）和AFP-L3的纯化和检测,并计算AFP-L3的百分含量后与患者的癌灶大小和肝癌分期进行相关性分析.结果 PHC患者血清AFP-L3百分含量与癌灶大小呈正相关（r=0.332,P=0.009）,与肝癌分期不相关（r=0.121,P=0.189）.结论 AFP-L3的测定对于判断癌灶的大小具有一定的临床意义.     

肝癌肝移植患者外周血中AFP mRNA和MXR7 mRNA的表达及其与预后的关系

目的： 探讨甲胎蛋白（ɑ-fetoprotein,AFP）mRNA和米托蒽醌抗性基因（Mitoxantrone-resistant 7，MXR7）mRNA作为游离癌细胞（isolated tumor cells,ITC）的标志物在肝癌肝移植患者外周血中的表达情况及其与术后肿瘤复发和转移的关系。方法: 以2002年4月至2003年12月期间的53例肝癌肝移植患者为对象，通过建立稳定可靠的实时荧光定量RT-PCR检测方法来定量检测肝癌患者外周血全细胞中AFP mRNA和MXR7 mRNA在整个肝移植术围手术期的表达和动态变化情况，并与术后肝癌复发和转移进行相关分析。 结果: 将研究对象分为肝癌移植组（53例）、晚期肝癌组（8例）、良性肝病组（26例）、正常对照组（10例），检测发现在肝移植术前，晚期肝癌组的AFP mRNA和MXR7 mRNA其表达率均达到100％,在肝癌移植组中表达率分别为57.7%和53.6%。2者均明显高于在良性肝病组（表达率为19.2%和0）和正常对照组（表达率均为0）（P<0.05）。同时在2者的阳性结果表达水平上，晚期肝癌组和肝癌组要明显高于良性肝病组和正常对照组(P<0.05)，前2者阳性结果定量水平达到1×10¹²copies/g RNA，是后2者定量水平的100－200倍。围手术期AFP mRNA和MXR7 mRNA的表达无论是表达率还是阳性结果表达水平，均以在术中的检测标本为最高，而术后1周为次之。术后出现2次以上的持续性的AFP mRNA和MXR7 mRNA的表达往往提示肝癌的复发和转移，较之移植术后没表达或一过性升高的病例，其差异有统计学意义（P<0.05），而术前和术中单次的AFP mRNA和MXR7 mRNA的表达与否则对术后肝癌复发和转移无明显影响（P>0.05）。结论: AFP mRNA和MXR7 mRNA可以作为肝癌ITC细胞的标志物，具有较好的特异性。术后血全细胞检测出现2次以上的持续性的AFP mRNA和MXR7 mRNA的表达可以有效预测肝癌肝移植术后肿瘤复发和转移。