高强度聚焦超声治疗对肝癌患者外周血AFP mRNA的影响

目的　探讨高强度聚焦超声治疗对肝癌细胞血道播散的影响。方法　应用Nested RT PCR技术检测19例患者高强度聚焦超声治疗前、后外周血AFPmRNA的表达 ,观察其对肝癌细胞血道播散的影响。结果　①高强度聚焦超声治疗前 ,19例肝癌患者外周血检出AFPmRNA表达者 11例 ( 5 7.9% ) ,良性疾病对照组均未检出。治疗前AFPmRNA表达阳性率与血清AFP水平、肿瘤直径、门静脉癌栓、远处转移等临床参数明显相关 (P<0 .0 5 )。②余 8例治疗前AFPmRNA表达阴性者 ,有 2例于术后即刻转为阳性 ,其中 1例于 72h后又复转为阴性 ;19例患者于治疗后 1周时其AFPmRNA表达阳性率 ( 31.6 % )低于治疗前 ,但差异无统计学意义 (P>0 .0 5 )。③治疗后 1周时 ,肿瘤直径≤8cm者AFPmRNA表达阳性率低于直径>8cm者 (P<0 .0 5 )。结论　高强度聚焦超声治疗可能会降低肝癌的血道播散 ,特别是对于肿瘤直径 ≤8cm的肝癌患者 ,疗效较好。     

MAGE-10基因在肝细胞肝癌中的表达及临床意义

目的　探讨MAGE - 10基因与肝细胞肝癌 (HCC)患者临床指标及转移复发的关系。　方法　用RT -PCR的方法测定 31例HCC癌组织及相应癌旁组织和 10例肝硬化组织MAGE - 10基因表达 ,对全部RT -PCR扩增产物中目的基因片段进行DNA测序以证实其为MAGE - 10基因 ,31例HCC测定AFP、岩藻糖苷酶 (AFU)、抗HCV、HBsAg、AFPmRNA、肿瘤直径等临床指标。　结果　 31例HCC组织中MAGE - 10基因表达的阳性率为 5 1 6 % (16 / 31) ,明显高于癌旁组织 0 % (0 / 31) ,(χ2 =18 95 ,P <0 0 1)。HCC组织中MAGE - 10基因表达的阳性率与血清AFP、AFU、抗HCV、HBsAg、AFPmRNA水平及肿瘤大小无关 (P >0 0 5 )。 10例肝硬化组织中未见MAGE - 10基因的表达。　结论　MAGE - 10基因在HCC组织中特异高表达 ,其阳性率与HCC患者肿瘤标志物无关。     

荧光定量检测肝癌、癌旁组织及外周血AFP mRNA

目的　探讨肝癌、癌旁组织及外周血AFPmRNA的表达意义。方法　应用荧光定量聚合酶链反应 (FQ -PCR)定量检测 30例原发性肝癌术前外周血、肝癌组织、癌旁组织AFPmRNA。结果　肝癌组织AFPmRNA的基因拷贝数与组织的分化程度、有无肝内浸润和 (或 )转移及血清AFP明显相关 ;而与外周血的AFPmRNA的表达相关无显著性 ;肝癌组织与癌旁组织AFPmRNA的表达量差异有显著性。结论　定量检测AFPmRNA表达情况可能判断肝癌组织的分化程度及早期诊断微小肝癌、肝癌的复发和转移 ,并指导治疗 ,判断预后。     

巢式RT-PCR定量检测肝癌、癌旁组织及正常肝组织内AFPmRNA

目的:探讨肝癌、癌旁组织及正常肝组织内AFPmRNA的表达含量 。方法:应用巢式RT-PCR定量检测肝癌、癌旁组织及正常肝组织内AFPmRNA 。结果:17例AFP（+）癌旁组织内AFPmRNA表达强阳性（+++）,其癌旁组织内AFPmRNA表达呈中度阳性（++）。15例AFP（-）肝癌、癌旁组织及正常肝组织内AFPmRNA表达弱阳性（+） 。结论:AFP（+）肝癌组织过量表达AFPmRNA基因,正常肝组织仅微弱表达AFPmRNA。     

肝癌患者不同部位血甲胎蛋白mRNA的检测及其临床意义

目的 检测肝癌患者不同部位（门静脉、肝静脉及外周静脉）血甲胎蛋白（AFP）mRNA的表达,了解肝细胞肝癌血行播散的规律。方法 采用巢式逆转录－聚合酶链式反应（RT－PCR）检测技术,测定20例肝癌患者门静脉、肝静脉及外周血AFPmRNA的表达。结果 全组病例门静脉、肝静脉血AFPmRNA阳性率（65％）高于其外周血AFPmRNA阳性率（40％－45％）,但差异无显著性（P＞0．05）。肿瘤≤5cm患者的肝静脉血AFPmRNA阳性率（70％）明显高于外周血（20％,P＜0．05）,门静脉血AFPmRNA阳性率（60％）也高于外周血（30％）,但差异无显著性（P＞0．05）。肿瘤＞5cm患者的门静脉、肝静脉血AFPmRNA阳性率接近于外周血AFPmRNA的阳性率。结论 肿瘤＞5cm的中晚期肝癌患者,其外周血中AFPmRNA的表达能较好地反映脱落癌细胞的血行播散情况;肿瘤≤5cm的患者虽然其外周血AFPmRNA阳性检出率较低,但肝静脉血的AFPmRNA明显高于外周血,提示肝内已有肿瘤播散,此时直接检测肝内门静脉或肝静脉血的AFPmRNA表达能更准确地反映癌细胞血行播散情况。     

外周血γ-谷氨酰转移酶mRNA-H亚型与甲胎蛋白对原发性肝癌诊断及肝外转移意义的比较

目的　探讨外周血γ 谷氨酰转移酶 (GGT)mRNA H亚型及甲胎蛋白 (AFP)的表达与原发性肝癌 (HCC)诊断及肝外转移的关系。方法　以逆转录 聚合酶链反应方法检测HCC及对照组外周血GGTmRNA H亚型 ;运用放射免疫测定法检测外周血AFP水平。结果　 65例肝癌中 ,GGTmRNA H亚型阳性率为 46.2 %。小肝癌AFP阳性率 (3 0 .0 % )明显低于大肝癌 (75 .6% ) ;其余各组AFP阳性率无差异 (P >0 .0 5 ) ;有肝内转移 (82 .1% )、门静脉癌栓 (86.4% )及远处转移组(90 .0 % )其外周血GGTmRNA H亚型阳性率显著高于无肝内转移 (18.9% )、门脉癌栓 (2 5 .6% )及远处转移组 (3 8.2 % ) (P <0 .0 5 )。血清AFP值的高低与肝癌患者外周血GGTmRNA H的阳性率两者差异无显著性 (P >0 .0 5 )。结论　检测HCC患者外周血细胞GGTmRNA H亚型表达对估计有无血行播散和远处转移有临床实用价值。对血清AFP值低或阴性患者进行周围血GGTmRNA H测定可以显著提高肝癌诊断的敏感性和特异性     

MAGE-1基因在肝细胞肝癌中的表达及临床意义

目的:研究MAGE-1基因在肝细胞肝癌组织中的表达,结合临床资料分析,探讨MAGE-1基因与肝细胞肝癌(HCC)患者临床指标及转移与复发的关系,为MAGE-1基因编码蛋白用于HCC患者免疫治疗提供依据。方法:用RT-PCR的方法对31例HCC患者癌组织及相应癌旁组织MAGE-1基因表达进行检测,随机对6例RT-PCR扩增产物中目的基因片段进行DNA测序以证实其为MAGE-1基因,所有患者均测定并统计AFP、AFU、抗HCV、HBsAAg、AFPmRNA、肿瘤直径等临床指标。结果:31例HCC患者肝癌组织中MAGE-1基因表达的阳性率64.5％(20/31)明显高于癌旁组织2％(1/31),P<0.01。HCC患者肝癌组织中MAGE-1基因表达的阳性率与患者AFP、AFU、抗HCV、乙肝标志物、AFPmRNA、肿瘤直径等临床指标均无关,P>0.05。结论:MAGE-1基因在HCC患者肝癌组织中特异高表达,HCC患者肝癌组织中MAGE-1基因表达的阳性率与HCC患者AFP、AFU、AFPmRNA肿瘤转移、复发均无关。     

肝细胞癌患者外周血单个核细胞端粒酶活性检测

目的　探讨外周血单个核细胞 (PBMC )端粒酶活性的检测在肝细胞癌 (HCC)患者中的意义。方法　采用聚合酶链反应 -酶联免疫吸附测定 (PCR ELISA )检测HCC、肝脏良性肿瘤、慢性肝病患者及正常人血PBMC和HCC组织端粒酶活性。结果　端粒酶活性在HCC患者的血PBMC和癌组织分别为 1.2 9± 0 .68和 1.40± 0 .88,取端粒酶活性 >0 .7为阳性标准 ,则HCC肝癌组织阳性表达的 2 8例全部为PBMC阳性表达 ,两者有显著一致性 (Kappa =1,P <0 .0 0 1)。血PBMC端粒酶活性在肝脏良性肿瘤组为 0 .65± 0 .3 2 ,慢性肝病组为 0 .43± 0 .2 3 ,正常对照组为 0 .3 8± 0 .14 ,均显著低于HCC组 (分别P <0 .0 1,<0 .0 0 1,<0 .0 0 1)。HCC中 ,在血清AFP高表达 (>2 0 0 μg/L )的18例中PBMC端粒酶活性全部为阳性 (10 0 % ) ,在血清AFP低表达 (<2 0 0 μg/L )的 12例中有 83 .3 % (10 /12 )PBMC端粒酶活性为阳性 ,差异有显著性 (P <0 .0 0 1)。且端粒酶活性的阳性率显著高于血清AFP (18/3 0 ,60 % ,P <0 .0 1)。结论　HCC患者的血PBMC端粒酶活性可准确反映癌组织端粒酶的表达情况 ,检测血PBMC端粒酶活性可能是一种灵敏、微创的早期诊断肝癌的有效方法。     

肝癌患者转化生长因子-β_1及其受体的表达和意义

目的　探讨肝癌患者转化生长因子 β1(TGF β1)及其受体的表达情况和意义。方法　酶联免疫吸附法检测 2 7例行胆囊切除术者 (对照组 )和 36例肝癌患者 (肝切除组 )肝切除手术前后血清TGF β1浓度 ;免疫组织化学法检测 36例肝癌和癌周组织标本中相关受体的表达情况。结果　肝癌患者血清TGF β1浓度较对照组显著升高 (P <0 .0 5 ) ;患者肝切除术后血清TGF β1浓度较术前有显著升高 (P <0 .0 1) ,而胆囊切除患者则无明显变化。癌组织与癌周组织受体表达的强度近似 ,但是部位有显著不同 ,36例癌周组织无论是否有肝硬化 ,受体染色均为阳性 ,以在细胞膜表面线形浓聚为特点 ,36例癌组织中仅 11例阳性 ,而且受体染色在细胞核周的胞浆中呈颗粒状浓聚 ,细胞膜表面则未发现受体染色。两者受体的染色阳性率差异有非常显著性 (P <0 .0 1)。结论　血清TGF β1水平可能是肝癌重要的血清标记物之一。肝叶切除能引起血清TGF β1水平的升高 ,但该因子受体只表达于癌细胞核周围的胞浆中 ,而在癌周组织中 ,则表达于细胞膜表面 ,提示癌细胞受TGF β1影响较小 ,故肝癌细胞的分裂增殖失控与TGF β1的抑制作用降低有关。     

多项肿瘤标志物联合检测对肝癌早期诊断的作用

目的 探讨多项肿瘤标志物联合检测在原发性肝癌中的诊断价值及建立判别方程.方法 采用蛋白芯片技术,检测2003年11月至2006年4月大坪医院收治的98例原发性肝癌患者(肝癌组)、67例良性肝病患者(肝病组)、46例健康体检者(对照组)血清中的12项肿瘤标志物,并在肝癌组与肝病组患者之间建立判别方程.采用方差分析和X2检验对检测结果进行分析.结果 肝癌组中87例患者肿瘤标志物呈阳性表达(89%),肝病组中有13例呈阳性表达(19%),对照组中有2例呈阳性表达(4%).3组中的AFP、CEA、铁蛋白、CA19-9和CA125检测结果比较,差异有统计学意义(F=59.530,40.472,31.708,75.897,153.066,P<0.05).联合检测这5项指标,肝癌临床诊断符合率提高为89%,明显高于单项AFP检测的64%(X2=16.362,P<0.05).所建判别方程的判断准确率为90%.结论 多项肿瘤标志物联合检测优于单独AFP检测,可用于对肝癌高危人群的筛查及原发性肝癌的早期诊断.     

测定胃液中AFP与CEA在胃癌诊断中的价值

目的探讨胃液中的AFP与CEA检测在胃部恶性肿瘤和胃部良性疾病诊断中的作用.方法应用放射免疫分析法,测定34位胃癌病人,22例胃部良性病患者的AFP与CEA的含量并进行对比.结果 34例胃癌病人胃液中的AFP与CEA的阳性检出率及联合诊断率均明显高于胃部良性疾病的患者水平(P＜0.005).结论测定病人胃液中的AFP与CEA的含量,特别是联合检测对胃癌的诊断和鉴别诊断有较高的临床价值.     

原发性肝癌患者介入治疗前后血清GPC3水平检测的临床价值

目的:了解原发性肝癌(hepatocellar carcinoma,HCC)介入治疗前后GPC3的浓度变化和临床价值.方法:采用ELISA法检测HCC、肝硬化、正常人血清GPC3浓度.结果:51例HCC患者、16例肝硬化患者、正常人组血清GPC3浓度分别为(3 939.72±2 801.92)、(619.08±542.80)和(534.24±500.20)pg/mL,前者与后二者比较差异有统计学意义(P<0.05),但其表达与血清甲胎蛋白(alpha-fetoprotein,AFP)无显著相关性;32例HCC患者介入治疗后高转移复发倾向组血清GPC3浓度为(4 418.00±3941.96)pg/mL,低转移复发倾向组血清GPC3浓度为(534.44±910.56)pg/mL,二者比较差异有统计学意义(P<0.05);缓解组、稳定组、恶化组血清GPC3浓度分别为(571.64±288.72)、(2 521.76±2340.76)、(4 812.84 4-1 858.64)pg/mL,3组两两比较差异均有统计学意义(P<0.05).结论:GPC3在HCC患者血清中的检出率高,特异性强,其表达与AFP无相关性,联合应用AFP及GPC3可提高诊断的灵敏度.GPC3与肿瘤复发转移、治疗效果有关.     

肝细胞癌组织中HBsAg、AFP的研究

本研究对65例肝细胞癌标本进行免疫组化测定组织细胞中的HBsAg和AFP,结果显示肝癌细胞中存在有HBsAg、AFP,二者都主要存在于细胞浆中,表示HBV·DNA可以表达HBsAg抗原,致使肝细胞产生AFP,促发肝细胞癌。肝细胞癌中AFP反映了肝癌细胞的分化状态,比血清AFP更具有特异性的肝癌标志物。     

围手术期检测肝细胞癌病人外周血AFP mRNA的临床意义

目的　检测围手术期肝细胞癌病人外周血AFPmRNA ,通过随访评估其预后价值。方法　通过巢式RT PCR的方法 ,在围手术期检测了 4 5例肝细胞癌病人外周血中AFPmRNA的表达 ,并进行了 1～ 2 0个月不等的随访。结果　在 4 5例肝细胞癌病人外周血中 ,AFPmRNA的检出率分别是术前 5 1.1% ,术后 7d 2 4 .4 % ,术后 2 8d 2 2 .2 %。多因素统计分析提示 ,术后 2 8d外周血检出AFPmRNA是提示病人复发的独立预后因素。结论　采用巢式RT PCR的方法检测肝细胞癌病人术后 2 8d外周血AFPmRNA有助于预测肝癌转移复发     

携带反义甲胎蛋白增强子/单纯疱疹病毒胸苷激酶基因的产病毒细胞系的建立及对肝癌细胞的靶向杀伤效应

目的建立对肝癌细胞具有靶向调控的高感染性和高分泌性产病毒细胞系。方法将重组反义甲胎蛋白增强子/单纯疱疹病毒胸苷激酶基因逆转录病毒载体(pL/TK/AFE/SN)转染至PA317包装细胞,经G418(400mg/L)筛选、病毒滴度测定。以病毒上清感染培养的Hep3B肝癌细胞和Hela宫颈癌细胞,并抽提细胞基因组DNA作Southernblot分析;观察GCV(100mg/L)对细胞生长的影响。结果转染pL/TK/AFE/SN的PA317细胞经G418筛选2周获阳性克隆;病毒滴度为5.4×104～3.8×105CFU/ml。Southern杂交证实前病毒已完整整合于PA317细胞,并稳定整合于Hep3B肝癌细胞和Hela宫颈癌细胞中。导入AFE/TK基因的Hep3B肝癌细胞经GCV作用后的存活率在2、4、6d时分别降低35%、87%、95%,生长明显受抑制(F=7.23,P<0.01)。结论携带AFE/TK基因的产病毒细胞系能够成功地将AFE/TK基因转移到感染细胞中。     

Results of retroperitoneal lymph node dissection after chemotherapy in patients with pure seminoma in the orchidectomy specimen but elevated serum α‐fetoprotein

OBJECTIVE

To determine the incidence of necrosis, teratoma, and active cancer in specimens at retroperitoneal lymph node dissection (RPLND) after chemotherapy in patients who presented with a pure seminoma primary tumour and an elevated serum α‐fetoprotein (AFP) level at diagnosis who underwent surgery.

PATIENTS AND METHODS

A retrospective review of the Indiana University testis cancer database from 1980 to 2004 was performed to identify all patients with metastasic germ cell cancer, pure seminoma in the orchidectomy specimen, and an elevated AFP level. In all, 42 patients were identified; two with nonseminomatous germ cell cancer in the contralateral testicle were excluded.

RESULTS

RPLND pathology in the 40 patients showed necrosis in 13 (32.5%), teratoma in 12 (30%), and cancer in 15 (37.5%). The histological subtype of the 15 cancer specimens at RPLND was pure seminoma in two, embryonal in three, yolk sac in seven, variant in one, and mixed elements in two (one with seminoma and yolk sac, and one with embryonal and variant). In all, 20 patients presented with a serum AFP level of <1000 µg/mL with nine (45%) having teratoma only in the retroperitoneum in contrast to 20 patients with an AFP level of >1000 µg/mL with three (15%) having teratoma only in the retroperitoneum (P = 0.04). The level of serum AFP did not predict for active cancer in the retroperitoneum (P = 0.5).

CONCLUSIONS

RPLND in patients after chemotherapy who presented with pure seminoma in the orchidectomy specimen and an elevated AFP do not have a high probability of finding necrosis only in the RPLND specimen. In fact, the prevalence of persistent cancer is higher compared with the general group of patients that have RPLND after chemotherapy.     

Primary endodermal sinus tumor of the prostate: report of a case

A 29-year-old man was admitted to the Department of Urology of the University of Padova with a diagnosis of prostatic adenocarcinoma. Cystourethrography showed a prostatic urethral obstruction, and CT scan demonstrated a low pelvic solid mass involving the prostate and the seminal vescicle. A radical prostatectomy was performed, and the prostate was found to be widely infiltrated by a neoplasia formed by tubules, papillary structures, and solid cords of cuboidal or flat elements, each having indistinct borders and scant cytoplasm, in a myxoid or fibrous stroma. "Schiller-Duval-like," PAS-diastase-resistant and alfa-fetoprotein hyaline bodies were present. An endodermal sinus (yolk-sac) tumor was diagnosed. There was infiltration of the periurethral prostatic ducts, prostatic urethra, and seminal vesicles and single lymph node metastasis. Testicular echotomography was negative. In spite of aggressive surgery and chemotherapy, the patient died 10 months after diagnosis. This is the third case in the literature after those presented by Benson and Michel.     

Serum alpha-fetoprotein subfractions in hepatic malignancies identified by different reactivities with concanvalin A,lentil lectin or phytohemagglutinin-E

Using a modified method of concanavalin A (Con A), lentil lectin (LCH) or phytohemagglutinin-E (PHA-E) affinity crossed-line immunoelectrophoresis (ACIE), we studied alpha-fetoprotein (AFP) subfractions in 69 sera, including 58 from patients with primary liver cancer and 11 from patients with hepatic metastasis of gastric cancer. We found that Con A non-reactive subfraction (type b) or LCH weakly-reactive subfraction (type B) was more frequently detected in metastatic liver cancer, as compared with liver cancer hepatoma. The amount of Con A non-reactive subfraction (type b) or of PHA-E reactive subfraction (type X) was significantly higher in case of metastatic liver cancer than in primary liver cancer. Since different affinities between AFP and lectins are due to the microheterogeneity in AFP sugar chain, our findings suggest that AFP in primary liver cancer and metastatic liver cancer is glycosylated in a different manner. It is also indicated that different patterns of AFP subfractions identified by the combination of Con A, LCH or PHA-E ACIE facilitate a differential diagnosis of these hepatic malignancies. These data were reported at the 22nd Congress of the Japan Society for Cancer Therapy, on September 26, 1984, in Tokyo, Japan     

Alpha-fetoprotein-producing esophageal adenocarcinoma: Report of a case

This paper describes a rare case of adenocarcinoma located in the middle portion of the esophagus with liver metastasis. An 80-year-old man was admitted to our hospital with dysphagia and vomiting, following which an upper gastrointestinal series and esophagoscopy located an elevated-type carcinoma in the middle thoracic esophagus. Computed tomography revealed an esophageal tumor invading the left atrium and aorta, and multiple intrathoracic lymph node swellings, and an ultrasonograph of the liver showed multiple liver metastases. The serum carcinoembryonic antigen, carbohydrate antigen 19-9, and squamous cell carcinoma-related antigen levels were normal, but the serum alpha-fetoprotein (AFP) level was 351.5 ng/ml. The patient died 124 days after undergoing an esophageal bypass operation. On post-mortem histological examination, the original esophageal tumor was diagnosed as a poorly differentiated adenocarcinoma without a squamous component and immunohistochemical staining for AFP showed positive granules in the cytoplasm. All the metastatic nodules, including the lymph nodes, liver, spleen, and lungs, showed the same histological type and AFP-staining pattern as the original esophageal tumor. To our knowledge, this is the first case of AFP-producing esophageal carcinoma to be reported in Japan.     

外周血GPC-3 mRNA在大鼠肝癌微转移中的意义

目的　在大鼠肝细胞癌模型中 ,通过检测外周血GPC 3mRNA ,探讨其与大鼠肝癌微转移的关系 ,为临床肝癌的早期检测奠定实验基础。方法　采用二乙基亚硝胺建立大鼠肝癌模型 ,提取外周血总RNA ,经逆转录后 ,用巢式PCR检测GPC 3mRNA和AFPmRNA ,比较二者在外周血中的阳性率与肝癌微转移的关系。结果　 75例成功构建的肝癌大鼠中 ,其中 2 5例单个结节肝癌中GPC 3mR NA阳性率为 32 % ,AFPmRNA阳性率为 2 8% ;2 0例 2～ 5个结节肝癌中GPC 3mRNA阳性率为 4 5 % ,AFPmRNA阳性率为 35 % ;18例大于 5个结节肝癌中GPC 3mRNA阳性率为 72 .2 % ,AFPmRNA阳性率为 6 1.1% ;12例其它器官有明显转移灶的肝癌中GPC 3mRNA阳性率为 83.3% ,AFPmRNA阳性率为75 % ;18例正常大鼠中GPC 3未检测到 ,而AFPmRNA阳性率为 11.1%。结论　随着肿瘤发生转移的可能性增大 ,GPC 3mRNA的阳性率明显增高 ,表明外周血GPC 3mRNA的阳性率与肝癌微转移的发生有明显的相关性 ,且高于AFPmRNA的阳性率。有望成为肝癌微转移新的诊断指标。