肝细胞癌与肝内胆管癌的免疫组化诊断

目的 评价一组免疫组化标志物在诊断和鉴别诊断肝细胞癌(HCC)和肝内胆管癌(ICC)中的价值.方法 对手术切除的90例HCC和80例ICC分别进行石蜡包埋肝细胞1(Hep Par 1)、多克隆性癌胚抗原(pCEA)、CD34、CD10、CD105、多药耐药相关蛋白3(MRP-3)、环氧合酶2(COX-2)、黏糖蛋白1(MUC-1)、水通道蛋白1(AQP-1)和CK19等10种抗体的免疫组化染色,比较其表达阳性率的差异性.结果 Hep Par 1、pCEA、CD34、CD10、CD105、MRP-3和COX-2在HCC的表达阳性率分别为85.6%、82.2%、87.8%、18.9%、8.9%、11.1%和48.9%,MUC-1、AQP-1和CK19在ICC的表达阳性率分别为73.8%、65%和92.5%.结论 HCC的一线诊断抗体由Hep Par 1和CD34组成,二线诊断抗体由pCEA和COX-2组成;ICC的一线诊断抗体由MUC-1和CK19组成,二线诊断抗体为AQP-1.     

肝肿瘤鉴别诊断中免疫组化应用的研究进展

论述肝细胞癌（HCC）器官特异性抗体HepPar-1、甲胎蛋白（AFP）,HCC相对特异性抗体多克隆癌胚抗原（pCEA）、氨肽酶N（CD13）、CD10（neprilysin）、绒毛蛋白、CD34,肝内胆管细胞癌（ICC）与转移性腺癌（MAC）相对特异性抗体细胞角蛋白（CK）7、CK19、CK20、MOC31、甲状腺转录因子-1（TTF-1）等单独及联合应用在肝肿瘤鉴别诊断中作用的研究进展。     

Combined hepatocellular andcholangiocarcinoma originating fromthe same clone：a pathomolecular evidence-based study

Background:Combined hepatocellular and cholangiocarcinoma (CHC) is a unique subtype of liver cancer com?prising both hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma (ICC); however, its cellular origin remains unclear. The purpose of this study was to investigate the clinicopathologic features and the clonal relation?ship between HCC and ICC in 34 patients with CHC. Methods:The clinicopathologic features and prognosis of the 34 CHC patients were compared with those of 29 patients with separated HCC and ICC (SHC). Loss of heterozygosity (LOH) at 10 highly polymorphic microsatellite markers was detected in 16 CHC and 10 SHC tissues for determination of the clonal origin of CHC. Expression of hepatocyte markers [hepatocyte paraffn 1 (Hep Par 1) and glypican 3 (GPC3)] and cholangiocyte markers [cytokeratin (CK)7 and 19] in tumor tissues was examined by immuno histochemical analysis. Results:In the 16 CHC specimens, the difference in LOH patterns between HCC and ICC was less than 30%, suggest?ing the same clonal origin of HCC and ICC. Consistent with this ifnding, immunohistochemical analysis revealed that hepatocyte markers (Hep Par 1 and GPC3) and cholangiocyte markers (CK7 and CK19) were simultaneously expressed in both the HCC and ICC components in 52.9% of CHC specimens, suggesting that the two components shared a similar phenotype with hepatic progenitor cells (HPCs). On the contrary, in all 10 SHC cases, the difference in LOH patterns between the HCC and ICC components was greater than 30%, suggesting different clonal origins of HCC and ICC. Overall survival and disease?free survival were shorter for patients with CHC than for patients with SHC (P<0.05). Conclusions:Our results suggest that the HCC and ICC components of CHC may originate from the same clone, hav?ing the potential for dual?directional differentiation similar to HPCs. CHC tended to exhibit the biological behaviors of both HCC and ICC, which may enhance the inifltrative capacity of tumor cells, leading to poor clinical outcomes for patients with CHC.     

肝脏肿瘤组织中HEP、AFP、CK19及CD1O的表达及意义

目的研究HEP、AFP、CK19及CD10在肝脏肿瘤组织中的表达及在鉴别诊断中的应用价值。方法对272例肝细胞肝癌（HCC）、62例胆管细胞癌（BCC）、27例混合性肝癌（CHC）、40例肝脏转移性癌（MC）及10例肝血管平滑肌脂肪瘤（AML）分别进行HEP、AFP、CK19及CD10免疫组织化学标记。结果HEP在HCC中表达率为91．91％，明显高于BCC（9．67％）、MC（12．5％）及AML（0％）（P〈0．01），且HEP表达与HCC的分化相关；AFP在HCC、BCC、CHC、MC及AML中表达率分别为47．79％（130／272），0％（0／62）、48．14％（13／27），10％（4／40），0％（0／10），其在HCC中的表达亦明显高于BCC、MC及AML（P〈0．01）；HCC及BCC中CK19的表达率分别为23．16％（63／272）、100％（62／62），后者明显高于前者（P〈0．01）；而CDl0在HCC中呈现特征性的细胞间小粱状、分支状及逗点状着色，阳性率为50．74％，而在BCC、MC及AML均无此着色。结论联合检测HEP、AFP、CK19及CD10对区别HCC及非HCC具有较高的应用价值。     

多肿瘤标志物蛋白芯片在卵巢癌诊断中的应用

目的：探讨多肿瘤标志物蛋白芯片技术对卵巢癌诊断的应用价值。方法：应用多肿瘤标志物蛋白芯片技术测定66例卵巢癌患者，64例妇科良性病患者和144例健康女性的血清12项肿瘤标志物（CA19-9、NSE、CEA、CA242、CA125、CA153、AFP、Ferritin、f-PSA、PSA、β-HCG、HGH）的水平，并评价蛋白芯片的诊断价值。结果：卵巢癌组的血清CA19-9、CEA、CA242、Ferritin、AFP、CA125和CA153水平显著高于妇科良性病组和健康对照组（P〈0．01）。卵巢癌组CA125、Ferritin、CA19-9、CEA、CA242和CA153单项指标的阳性率依次分别为68．2％、31．8％、30．3％、22．7％、19．7％和19．7％，与妇科良性病组和健康组比较有显著性差异（P〈0．001）。蛋白芯片12项指标联合检测的阳性率显著高于任何单项标志物（P〈0．001）。蛋白芯片检测中CA125＋CA19-9二项和CA125＋CA19-9＋CEA三项联合检测的诊断敏感性和准确性明显高于其CA125单项指标。蛋白芯片12项指标联合检测的诊断敏感性、特异性、准确性、阳性预测值和阴性预测值分别为87．9％、93．3％、92．0％、80．6％和96．0％。与其CA125单项指标比较，诊断敏感性和准确性分别从68．2％和90．5％增加到87．9％和92．0％。结论：应用蛋白芯片技术联合检测肿瘤标志物有效提高诊断敏感性和准确性，对卵巢癌的辅助诊断具有重要临床应用价值。     

CD147和CK19在肝细胞癌中的表达及临床意义

目的 探讨细胞外基质蛋白诱导因子（CD147）和细胞角蛋白19（CK19）在肝细胞癌（HCC）中的表达及临床意义。方法 采用组织芯片技术和免疫组织化学法检测CD147和CK19在272例HCC组织和81例癌旁组织中的表达情况。结果 CD147在HCC中的阳性表达率为73.53％（200／272）,在癌旁组织中的阳性表达率为13.58％（11／81）,差异有统计学意义（P＜0.05）。CK19在HCC组织中的阳性表达率为14.34％（39／272）,CK19在癌旁组织中无表达。在HCC中,CD147的表达与组织学分级、临床分期、术后无瘤生存时间、肿瘤直径、脉管或门静脉癌栓相关,与性别、年龄、肝硬化、AFP水平、HBV感染、淋巴结转移、病灶数目、肝被膜浸润及卫星灶无关（P＞0.05）;CK19在HCC中的表达与术后无瘤生存时间、组织学分级、肿瘤直径、肝硬化、卫星灶、淋巴结转移及临床分期有关,与性别、年龄、病灶数目、肝被膜浸润、AFP水平、HBV感染及脉管或门静脉癌栓无关（P＞0.05）。在HCC中,CD147 阳性表达者与阴性表达者的中位复发时间分别为13个月和48个月（P＜0.05）,中位生存时间分别为24个月和60个月（P＜0.05）;CK19阳性表达者与阴性表达者的中位复发时间分别为7个月和31个月（P＜0.05）,中位生存时间分别为13个月和42个月（P＜0.05）。 CD147的表达和CK19的表达无明显相关性（r=0.061,P=0.317）。结论 HCC中CD147和CK19的表达与预后密切相关,且两者均可作为HCC预后不良的判断指标。     

β-HCG与肝细胞癌的相关性研究

目的评估人绒毛膜促性腺激素（β-HCG）检测在肝细胞癌（HCC）诊断和预后中的价值。方法用微粒子酶免发光技术（MEIA）测定83例HCC患者，25例肝硬化患者和62名正常人血清的5种HCC血清标志物，包括β-HCG、甲胎蛋白（AFP）、糖链抗原19—9（CA19—9）、糖链抗原125（CA125）、癌胚抗原（CEA）。结果通过5种血清学指标检测，HCC患者5种血清学指标与肝硬化组和正常人组比较差异均有显著性（P〈0．01），β-HCG和AFP在Ⅰ期HCC和Ⅰ、Ⅲ期HCC中表达差异无显著性。部分AFP阴性患者β-HCG可阳性。结论β-HCG检测可提高HCC早期诊断率，同时，也可能为HCC生物治疗提供一个新的靶点。     

上消化道恶性肿瘤患者血清多项肿瘤标志物联合检测的临床意义

目的评价癌胚抗原（CEA）、唾液酸化岩藻戊糖（CA19—9）、胃肠道癌抗原（CA242）、甲胎蛋白（AFP）、胃癌及卵巢癌抗原（CA724）、鳞状上皮细胞癌抗原（SCC）、组织多肽抗原（TPA）、细胞角蛋白（TPS）8项标志物联合检测,对上消化道恶性肿瘤的临床诊断、评价疗效及预后等方面的临床意义。方法对373例上消化道恶性肿瘤患者及50名健康体检者血清标本进行8项标志物检测,评价标志物水平与病情的关系。结果8项标志物诊断阳性率分别为：CEA26．80％,CA19—927．34％,CA24234．14％,AFP2．84％,SCC19．72％,CA72412．13％,TPA34．15％,TPS30．89％。8项标志物联合检查的总阳性率为89．05％。术前CEA、CA242、SCC阳性患者的生存时间较短。上消化道恶性肿瘤患者血清CA242和CA19—9水平呈密切的正相关。结论多项标志物联合检测在上消化道恶性肿瘤不同病理分型及临床分期中差异有统计学意义,且阳性率远高于任何标志物单独检测的结果。CA19—9与CA242可能成为监测病情的最佳组合。CA724、CA242和SCC3项标志物可分别作为贲门、胃、食管恶性肿瘤患者的不良预后指标。     

GPC3、Hep-Par-1在肝细胞癌中的表达及其价值

目的探讨磷脂酰肌醇蛋白聚糖3(GPC3)、Hep-Par-1在原发性肝细胞癌(HCC)诊断与鉴别诊断中的应用价值。方法应用免疫组化EliVision法,对已确诊的64例HCC、12例肝内胆管细胞癌(ICC)、11例肝转移性腺癌(MAC)、6例局灶结节性增生(FNH)、2例肝细胞腺瘤(HA)及28例癌旁肝组织进行GPC3、Hep-Par-1抗体标记,检测2种抗体在上述组织中的表达情况。结果 GPC3在HCC中表达率为91%(58/64),其中75%(48/64)为强阳性(+++),在其他组织中无表达,表达差异具有统计学意义(P<0.05)。Hep-Par-1在HCC中表达率为81%(52/64),在FNH、HA及癌旁肝组织中表达率均为100%,在MAC中表达率为9%(1/11),在其余组织中无表达,表达差异具有统计学意义(P<0.05)。结论 GPC3和Hep-Par-1对HCC的诊断均具有较高的敏感性及特异性,GPC3可取代Hep-Par-1成为HCC诊断的一线抗体。     

Immunohistological evaluation of single small hepatocellular carcinoma with negative staining of monoclonal antibody Hepatocyte Paraffin 1

BACKGROUND AND OBJECTIVES: The sensitivity and specificity of the monoclonal antibody Hepatocyte Paraffin 1 (Hep Par 1) for hepatocellular carcinoma (HCC) are very high, and the usefulness for differential diagnosis of hepatic tumors has been reported. However, there are some cases of HCC with negative staining for Hep Par 1. We examined the histopathological features of HCC with negative staining for Hep Par 1. METHODS: We examined 69 samples of single nodular HCC less than 2 cm in greatest dimension, resected from 1985 to 1994 in our hospital, with immunohistological staining for Hep Par 1, cytokeratin 19 (CK 19), MUC-1 glycoprotein (MUC-1), and epithelial membrane antigen (EMA). RESULTS: Hep Par 1 staining was positive in 64 cases (93%) and negative in 5 cases (7%). With regard to the histological structure, 3 of the 5 negative cases were scirrhous HCC. With regard to the grade of histological differentiation, 2 cases were poorly differentiated HCC, 3 cases were moderately differentiated HCC, and no well-differentiated HCC was found in the negative cases. CK 19, MUC-1, and EMA staining were negative in all cases. CONCLUSIONS: It is necessary to recognize the existence of Hep Par 1 negative HCC, in particular scirrhous HCC. This may be due to a different mechanism in the earlier stage of hepatocarcinogenesis.     

胸腹水细胞块的免疫细胞化学研究

[目的]行多项胸腹水细胞块的免疫细胞化学检测,探索一组鉴别良恶性及肿瘤起源的有价值的常规一抗试剂组.[方法]收集胸腹水标本制成细胞块,HE染色筛检出间皮细胞反应性增生及可疑恶性或查见恶性细胞的病例59例.免疫细胞化学方法采用SP法,一抗用HBME-1、钙网膜蛋白(CR)、E-cad、CD44、CK7、CK20,腹水加做CA19-9(女性加做CA125),胸水加做TTF-1.[结果]HBME-1在间皮瘤中表达57.1%(4/7)、转移腺癌中表达51.1%(23/45);钙网膜蛋白在间皮瘤中表达100%、腺癌中未表达;E-cad( )见于96.4%(53/55)恶性肿瘤;CD44( )见于反应性增生及恶性间皮瘤;TTF-1在肺癌中表达80.6%(25/31)、非肺源性未见表达;CK7( )在转移腺癌中表达86.7%(39/45),无特异性;CK20( )在肠癌中表达100%,CK7(-)/CK20( )具肠源性特异性;CA19-9在胃肠癌中表达100%,间皮瘤中亦表达2/7;CA125在卵巢癌表达75.0%(3/4),特异性100%.[结论]E-cad鉴别良恶性胸腹水;CR鉴别是否间皮起源、TTF-1鉴别肺源性、CK7/CK20鉴别肠源性转移癌具有特异性及敏感性特点,它们可作为常规一抗鉴别良恶性及肿瘤起源.CA125鉴别卵巢癌具有相同特点,可作为女性患者腹水常规一抗.     

162例胃肠道间质瘤的临床病理学及免疫表型特征

目的阐明胃肠道间质瘤的临床病理学与免疫表型特征。方法收集１６２例胃肠道间质瘤临床病理学资料,其中４６例进行免疫组织化学染色。标记抗体为波形蛋白（ｖｉｍｅｎｔｉｎ）、ＣＤ３４、肌特异性肌纤蛋白（ＭＳＡ）、平滑肌特异性肌纤蛋白（ＳＭＡ）、Ｓ１００蛋白、神经特异性烯醇化酶（ＮＳＥ）、突触素（ＳＹＮ）、细胞角蛋白（ＣＫ）、癌胚抗原（ＣＥＡ）、白细胞共同抗原（ＬＣＡ）等１０种抗体。结果病理学上肿瘤起源于胃肠道肌层,直径为０．５～４３ｃｍ,５５％为恶性。免疫表型特征为ｖｉｍｅｎｔｉｎ,占１００％、ＣＤ３４,占６４％、ＭＳＡ,占４７％、ＳＭＡ,占４１％、ＮＳＥ,占６１％、Ｓ１００蛋白,占１９％、ＳＹＮ,占１５％。胃肠道间质瘤不表达ＣＫ、ＣＥＡ、ＬＣＡ。结论间质瘤为起源于胃肠道肌层的最常见间叶性肿瘤,消化道出血与腹部包块为最常见症状。最常见的细胞学特征为梭形细胞和上皮样细胞。肿瘤的大小、核分裂相是关系到肿瘤良恶性和患者预后、生存的最重要因素。免疫组化证实仅有部分肿瘤具有不完全的平滑肌、神经或双向分化特征。     

Serum CA 19-9 and alpha-fetoprotein levels in primary hepatocellular carcinoma and liver cirrhosis.

Serum CA 19-9 and alpha-fetoprotein (AFP) levels were determined in 211 patients with liver cirrhosis and 27 with primary hepatocellular carcinoma (HCC) associated with liver cirrhosis. This was done to determine the usefulness of CA 19-9 level with respect to AFP level in distinguishing between these two illnesses, and to assess the influence of some clinical and biochemical variables on these tests in patients with liver cirrhosis with or without primary HCC. Pathologic AFP values were found in 23 of 27 (sensitivity, 85%) patients with HCC; CA 19-9 levels increased in only 12 of 27 (sensitivity, 44%) HCC patients, the values being comparable with those of patients with liver cirrhosis. In liver cirrhosis a substantial number of false-positive values was found for both markers, although they were higher for CA 19-9 (50 of 211 versus 39 of 211). In liver cirrhosis correlations were found between AFP level and alanine amino-transferase level; and between CA 19-9 level and (1) total bilirubin value, (2) alkaline phosphatase level, and (3) pseudocholinesterase level. The authors conclude that CA 19-9 level is a poor biochemical marker, inferior to AFP level, in the detection of a carcinomatous transformation of liver cirrhosis. The finding of false-positive AFP values in liver cirrhosis seems mainly attributable to cellular proliferation and necrosis. Cholestasis seems to greatly affect serum CA 19-9 level variations, probably by reducing its liver metabolism.     

Primary clear cell adenocarcinoma of the lung

Primary clear cell adenocarcinoma of the lung is extremely rare. A 63-year-old Japanese man consulted to our hospital because of cough and sputum. Imaging modalities including XP, CT and MRI revealed a tumor of the right middle lobe. They did not identify other tumors in the body. Because clinical cytology and biopsy showed malignant cells, segmentectomy of the lobe was performed. Grossly, the tumor was whitish tan tumor measuring 5 × 6 × 5 cm. Histologically, the tumor was composed entirely of clear cells arranged in papillary and tubular patterns. The tumor cells had hyperchromatic nuclei, and mitotic figure and nuclear stratification were scattered. Histochemically, glycogen and mucins were absent in tumor cell cytoplasm. Immunohistochemically, the tumor cells were positive for pancytokeratin (AE1/3, CAM5.2), cytokeratin (CK) 7, CK 8, CK18, CK19, EMA, CEA, CA19-9, CA125, p53, Ki-67 (labeling = 80%), TTF-1 and surfactant apoprotein A. In contrast, the tumor cells were negative for CK5/6, CK 34βE12, CK 14, CK 20, vimentin, desmin, S100 protein, α-smooth muscle actin, AFP, HMB45, CD10, CD34, HER2/neu, CD56, p63, and synaptophysin. The tumor recurred 6 months after the operation, and right middle lobectomy was performed. Postoperative imaging modalities showed no tumors. The patient is alive free from tumor 9 years after the first manifestation.     

Application of Joint Detection of AFP,CA19-9, CA125 and CEA in Identification and Diagnosis of Cholangiocarcinoma

Objective: To explore the application of joint detection of serum AFP, CA19-9, CA125 and CEA in identificationand diagnosis of cholangiocarcinoma (CC). Materials and Methods: The levels of serum AFP, CA19-9, CA125and CEA of both 30 patients with CC and 30 patients with hepatocellular carcinoma (HCC) were assessed.Receiver operating characteristic (ROC) curves were used to evaluate the diagnostic effects of single and jointdetection of those 4 kinds of tumor markers for CC. Results: The levels of serum CA19-9, CA125 and CEAin CC patients were higher than that in HCC patients,whereas that of serum AFP was significantly lower s.The area under ROC curve of single detection of serum AFP, CA19-9, CA125 and CEA were 0.05, 0.86, 0.84and 0.83, with the optimal cutoff values of 15.4 ng/ml, 125.1 U/ml, 95.7 U/ml and 25.9 ng/ml, correspondingly,and the percentage correct single diagnosis was <79%. With joint detection, the diagnostic effect of combinedAFP, CA19-9, CA125 and CEA was the highest, with an area under the ROC curve of 0.94 (95%CI 0.88~0.99).Conclusions: Single detection of serum CA19-9, CA125 and EA is not meaningful. The sensitivity, specificity,the rate of correct diagnosis and the area under ROC curve of joint detection of AFP, CA19-9, CA125 and CEAare highest, indicating that the joint detection of these 4 tumor markers is of great importance in the diagnosisof CC.     

血小板及其参数联合肿瘤标志物术前鉴别肝细胞癌及肝内胆管癌的临床价值

目的 探讨血小板(PLT)及其参数和甲胎蛋白(AFP)、糖类抗原199(CA199)、糖类抗原125(CA125)及癌胚抗原(CEA)在术前肝细胞癌(HCC)及肝内胆管癌(ICC)鉴别诊断中的价值.方法 回顾性分析兰州大学第二医院行手术治疗的肝癌患者274例,据术后病理将其分为HCC组229例和ICC组45例.比较两组...     

Stereotactic radiotherapy for hepatocellular carcinoma induced by hepatitis C and the relationships of changes in carbohydrate antigen 19-9 with AFP and PIVKA-II

PurposeAssessing the therapeutic effects of stereotactic body radiotherapy (SBRT) for hepatocellular carcinoma (HCC) takes time. Purpose of our study was to explore the relationships of changes in carbohydrate antigen 19-9 (CA 19-9) with those in the existing markers alpha-fetoprotein (AFP) and protein induced by vitamin K absence or antagonist-II (PIVKA-II).Patients and methodsThe subjects were 16 patients who underwent SBRT for solitary HCC ≤ 3 cm induced by hepatitis C between June 2016 and July 2019. Observation periods ranged from 8–43 (median: 28) months, ages from 59–85 (median: 65) years.ResultsChanges in CA 19-9 levels after SBRT were categorised into three patterns: 1) a transient elevation followed by a decline (75%); 2) a transient decline followed by an elevation (18.8%); and 3) no change (6.3%). Among patients showing a transient CA 19-9 elevation followed by a decline, which was the most frequent pattern, 75% showed these changes in synchronisation with AFP and preceded the changes in PIVKA-II, while in the other 25%, CA 19-9 changes were in synchronisation with PIVKA-II and preceded those in AFP. At the time of recurrence, 62.5% showed a continuous CA 19-9 elevation, either in synchronisation with other markers or by itself.ConclusionsThis is the first investigation of changes in CA 19-9 levels after SBRT for HCC induced by hepatitis C. Characteristic changes in CA 19-9, AFP, and PIVKA-II levels were observed as responses after treatment. As for its correlations with tumour markers, the acute responses of PIVKA-II tended to be slower than those of CA 19-9 and AFP. Although the sample size was small, our findings raise the possibility that measuring these 3 biomarkers after SBRT may be useful for monitoring patients for HCC recurrence.