肝脏局灶性结节性增生的诊断与治疗

目的:探讨肝脏局灶性结节性增生(focal nodular hyperplasia,FNH)的临床诊断和治疗。方法:回顾性分析18例经病理组织学检查确诊的FNH的临床病理、影像学检查、外科治疗及随访资料。结果:18例FNH,影像学检查诊断率较低,其中彩超确诊率44.4%(8/18),CT66.7%(12/18),MRI88.8%(16/18),手术切除病灶效果好,随访1年未发现复发病例。结论:目前CT和MRI是FNH诊断的重要手段,对影像检查疑为FNH又暂不同意手术探查病人可考虑定位穿刺活组织检查,但对穿刺未能定性成功及不能排除恶性肿瘤者应手术治疗。     

肝局灶性结节增生与肝血管瘤的超声造影鉴别

目的探讨超声造影对肝局灶性结节增生和肝血管瘤的清晰程度。方法经增强CT或MR对照,并经手术及病理证实的肝局灶性结节增生病变患者10例,肝血管瘤患者21例。采用第二代超声造影剂SonoVue进行低机械指数实时超声造影成像,记录肝实质及病灶的充填过程、充盈方式及增强程度。结果肝局灶性结节增生和肝血管瘤的造影模式各有其特点,其主要鉴别点在于病灶的充填过程及充填方式,10例肝局灶性结节增生动脉相均为快速增强,其中7例为中央型,3例为整体型,门脉相均为高回声,延迟相8例为高回声,2例为等回声;21例肝血管瘤,动脉相表现为周边环状增强,中央无增强,门脉相继续由周边环状或结节状向中央向心性增强,延迟相均呈整体增强。结论超声造影能清楚显示肝局灶性结节增生和肝血管瘤的造影充填过程及充填方式,对肝局灶性结节增生和肝血管瘤的鉴别诊断具有重要的意义。     

克隆性分析在肝细胞腺瘤与局灶性结节性增生鉴别诊断中的应用

目的 探讨肝细胞腺瘤（HA）和局灶性结节性增生（FNH）的克隆构成特征及其在两者鉴别诊断中的应用。方法 2例女性HA标本和发生于3位女性患者的4个FNH标本均经中性福尔马林固定、石蜡包埋、HE染色后应用显微切割技术分离病变及病变周围肝实质,提取基因组DNA,经甲基化敏感的HpaⅡ或Hha Ⅰ消化,巢式PCR扩增磷酸甘油酸激酶（PGK）和雄激素受体（AR）基因。通过Bst Ⅺ消化和琼脂糖电泳显示PGK基因单核苷酸多态性;应用变性聚丙烯酰胺凝胶电泳显示AR基因CAG重复序列长度多态性。选取4例高分化肝细胞癌（HCC）作为对照。结果 2例HA光镜下细胞排列成条索状,厚约2层,局部伴有脂肪变性及糖原贮积。瘤细胞呈圆形或多角形,大小和形态与周围肝细胞相似,未见核分裂象。克隆性分析结果显示,2例HA病变及4例HCC均显示出X染色体失活嵌合性丢失,证明为肿瘤性病变。4个FNH病变均缺乏特征性的中央瘢痕,整个病变显示多克隆性细胞组成。结论 HA属于单克隆性,诊断中需要与FNH及高分化HCC鉴别,除组织学观察外,克隆性分析在肝细胞腺瘤与结构不典型的FNH病变的鉴别诊断中有重要应用价值。     

HSP70、GPC3、GS和AKR1B10在肝脏高度异型增生结节免疫组化诊断中的应用

目的探讨免疫组化标志物热休克蛋白70(heat shock protein 70,HSP70)、磷脂酰肌醇蛋白聚糖3(glypican 3,GPC3)、谷氨酰胺酶(glutamine synthetase,GS)和正醛酮还原酶家族中单分子醛糖还原酶(aldo-ketoreductase family 1 member B10,AKR1B10)在肝脏高度异型增生结节(high-grade dysplastic nodule,HGDN)和高分化小肝细胞癌(well-differentiated small hepatocellular carcinoma,WD-SHCC)中的表达特点及鉴别诊断价值。方法对16例单结节型HGDN和32例WD-SHCC进行HSP70、GPC3、GS和AKR1B10免疫组化染色。结果 4项标志物在HGDN与WD-SHCC组织中均可表达,单项标志物中,HSP70在HGDN与WD-SHCC中阳性率最高(31.25%,81.25%,P0.001);GPC3在HGDN与WD-SHCC中阳性率最低(12.50%,25.00%,P0.460)。HSP70+GPC3+GS及HSP70+AKR1B10+GS诊断组合均在3项标志物中至少2项阳性时,获取最佳诊断效果,此时诊断敏感性和准确率分别为62.50%、81.25%和70.83%、83.33%,特异性均为87.50%。结论HSP70、GPC3和GS经典免疫组化诊断谱对HGDN和WD-SHCC组织具有一定的鉴别能力。然而,GPC3在WD-SHCC中阳性率较低,表达水平与HGDN无明显差异,制约该谱的整体诊断效果。AKR1B10替代GPC3后,在维持较高特异性一致的同时,可明显提高诊断敏感性和准确率。     

超声造影在肝硬化背景-F鉴别增生性结节与肝细胞性肝癌的应用价值

目的:探讨超声造影(contrast-enhanced ultrflBound,CEUS)在肝细胞性肝癌(hepatocellular caFcino-ma,HCC)与肝硬化增生结节诊断和鉴别诊断中的应用价值.方法:采用Sono Vue实时灰阶超声造影,对32例肝硬化背景下的16个肝细胞性肝癌结节与29个肝硬化增生性结节进行超声造影检查,造影时记录并观察其动脉相、门脉相和实质相的动态造影变化及TIC曲线变化,并与手术或穿刺病理诊断结果进行对照.结果:肝细胞性肝癌患者中93.75%(15/16)的病灶具有造影剂的快进快出的特点,超声造影剂在肝细胞性肝癌中快进快出,即动脉相快速增强,门脉相或实质相快速消退.肝硬化增生结节患者中89.66%(26/29)造影后可根据动脉期无明显增强,实质期无明显退出的诊断标准排除恶性.结论:超声造影在肝硬化增生结节与肝细胞性肝癌的鉴别诊断中具有重要的临床意义和应用价值.     

血管紧张素转换酶Ⅰ在肝局灶性结节状增生中的表达及其意义

收集四川大学华西医院病理科1993年5月-2005年3月确诊为肝局灶性结节状增生（FNH）的20例资料,包括症状和体征、实验室和影像学检查,术中所见以及治疗等情况,并进行随访。其中结节性肝硬化6例,肝细胞腺瘤5例;正常肝脏组织7例（其中2例为尸体解剖标本）。20例FNH的组织样本经4%甲醛固定,常规石蜡包埋切片,HE染色,光镜观察。     

超声造影在肝硬化背景下鉴别增生性结节与肝细胞性肝癌的应用价值

目的：探讨超声造影(contrast-enhanced ultrasound,CEUS)在肝细胞性肝癌(hepatocellular carcinoma，HCC)与肝硬化增生结节诊断和鉴别诊断中的应用价值。方法：采用Sono Vue 实时灰阶超声造影,对32例肝硬化背景下的16个肝细胞性肝癌结节与29个肝硬化增生性结节进行超声造影检查，造影时记录并观察其动脉相、门脉相和实质相的动态造影变化及TIC曲线变化，并与手术或穿刺病理诊断结果进行对照。结果：肝细胞性肝癌患者中93.75%(15/16)的病灶具有造影剂的快进快出的特点,超声造影剂在肝细胞性肝癌中快进快出，即动脉相快速增强,门脉相或实质相快速消退。肝硬化增生结节患者中89.66%(26/29)造影后可根据动脉期无明显增强,实质期无明显退出的诊断标准排除恶性。结论：超声造影在肝硬化增生结节与肝细胞性肝癌的鉴别诊断中具有重要的临床意义和应用价值。     

血清GPC3联合DCP、GP73及AFP检测在原发性肝癌诊疗中的临床应用

目的 探讨血清磷脂酰肌醇蛋白聚糖-3(GPC3)、异常凝血酶原(DCP)、高尔基体糖蛋白(GP73)及甲胎蛋白(AFP)联合检测在原发性肝癌(HCC)诊疗中的临床价值.方法 采用酶联免疫吸附试验(ELISA)法检测51例HCC患者血清中GPC3及DCP、GP73、AFP水平,并与36例肝硬化患者、63例肝纤维化患者、73例慢性活动性肝炎患者及80例表观健康人的结果进行比较,并比较其在HCC不同分期、分化程度和手术前后的表达情况.结果 HCC组血清GPC3、DCP、GP73及AFP的水平与肝硬化组、肝纤维化组、肝炎组和健康对照组比较,差异均有统计学意义(P<0.05);4个项目在HCC晚期(T3+T4期)水平均高于早期(T1+T2期),GP73和AFP在HCC晚期与早期的差异均有统计学意义(P<0.05),但GPC3和DCP在HCC晚期与早期的差异无统计学意义(P>0.05).GPC3、DCP、GP73及AFP在高、中、低分化HCC的水平变化不一,但4个项目在各分化程度之间的差异均无统计学意义(P均>0.05).与手术前相比,27例HCC患者手术后GPC3、DCP水平下降,GP73、AFP水平升高,GPC3水平差异有统计学意义(P<0.05),而DCP、GP73、AFP水平差异均无统计学意义(P均>0.05).结论 血清GPC3、DCP、GP73及AFP具有诊断和鉴别诊断HCC的价值,并对HCC患者的预后评估有一定的意义.     

Glypican-3和CD34免疫组化染色在肝细胞癌与良性肝细胞性病变鉴别诊断中的价值

肝细胞癌与良性肝细胞性疾病的鉴别诊断有时非常困难，特别是在细针穿刺活检时。作者应用免疫组化方法研究了107例肝细胞癌（HCC）、19例肝细胞腺瘤（HA）、16例局灶性结节性增生（FNH）和225例伴有上皮样分化的非肝脏性肿瘤中Glypican-3（GPC3）和CD34表达，结果发现88％（94／107）HCC胞质表达GPC3，而所有HA和FNH均不表达GPC3。     

Immunohistochemical detection of hcv in cirrhosis, dysplastic nodules, and hepatocellular carcinomas with parallel-tissue quantitative RT-PCR.

Hepatitis C virus is a major risk factor for hepatocarcinogenesis in humans. In situ detection of the virus in early sequential lesions of hepatocarcinogenesis could provide information about the role of the virus in the transformation and promotion process. Parallel in situ detection of HCV proteins and RNA in human tissues were performed in 55 posthepatitis C cirrhosis, 17 dysplastic nodules (DN), and 25 hepatocellular carcinomas (HCC), using immunohistochemistry and tissue quantitative RT-PCR. A consistent cytoplasmic hepatocellular staining was obtained in 73% of cirrhosis cases (with or without HCC) and in 55% DN cases. A few tumoral hepatocytes were unambiguously stained in 28% HCC. The percentage of positive cells and the intensity of immunostaining significantly decreased from cirrhosis to HCC through DN, whereas there was no difference in the prevalence of positivity or the number of viral copies between cirrhosis and HCC using tissue-quantitative RT-PCR. Finally, RT-PCR levels were found parallel with the immunostaining in cirrhosis but not in HCC. These results suggest that HCV protein synthesis may persist but be down-regulated during sequential hepatocarcinogenesis. A putative role of HCV proteins on cell proliferation and differentiation during the early steps of carcinogenesis cannot therefore be excluded.     

Promoter methylation of E-cadherin in hepatocellular carcinomas and dysplastic nodules

In order to clarify the significance of E-cadherin methylation in multistep hepatocarcinogenesis, we examined the methylation status of the E-cadherin promoter region, using methylation-specific polymerase chain reaction in 64 hepatocellular carcinomas (HCCs) and 13 dysplastic nodules (DNs), and correlated these results with E-cadherin protein expression and clinicopathologic factors of HCCs. Promoter methylation was detected in 1 of 13 (7.7%) DNs, in 5 of 13 (38.5%) Edmondson and Steiner grade I HCCs, and in 27 of 51 (52.9%) grade II or III HCCs, and a significant correlation was observed between the methylation status and the stepwise progression of hepatocarcinogenesis (p=0.004). Reduced E-cadherin immunoreactivity was found in 18 of 64 (28%) HCCs, but in none of DNs. E-cadherin methylation status in HCCs was significantly correlated with microvascular invasion (p=0.02) and tumor recurrence (p=0.04), but not with reduced E-cadherin immunoreactivity. The Kaplan-Meier method showed that methylation status did not have a significant influence on the recurrence-free survival of HCC patients (p=0.15). Our results indicate that methylation of the E-cadherin promoter region is a frequent event in HCC, which may play an important role in the stepwise progression of hepatocarcinogenesis. And the promoter methylation of E-cadherin in HCC was found to be significantly correlated with microvascular invasion and recurrence.     

Glypican-3 expression in hepatocellular tumors: diagnostic value for preneoplastic lesions and hepatocellular carcinomas

Glypican-3 (GPC3), a member of heparan sulfate proteoglycans, plays a role in cell growth, differentiation, and migration. The objectives of this study were to assess the diagnostic value of GPC3 immunostaining in hepatocellular carcinomas (HCCs) and to analyze its expression profile in preneoplastic lesions. Tissue microarrays were built by sampling 54 HCCs and adjacent liver tissues (21 developing from cirrhosis and 33 from normal liver) and 94 cirrhotic macronodules. Fourteen typical liver cell adenomas and 5 with malignant foci were also included. Sections were assessed for GPC3 expression by immunohistochemistry. GPC3 staining was observed in 19 (90%) of 21 HCC cases with cirrhosis and in 18 (64%) of 28 HCC cases with normal liver (P < .01). When staining was positive, it was both membranous and cytoplasmic. Positive staining was observed in 1 case of nonneoplastic adjacent liver. In cases of adenomas, only malignant foci were positive. Among the 94 macronodules, GPC3 immunostaining was noted in 48% (14/29) of high-grade dysplastic or early HCC and in 3% (2/65, P < .001) of benign or low-grade dysplastic macronodules. This study shows that GPC3 is an efficient diagnostic marker of HCC, potentially useful in the differential diagnosis of liver cell adenomas and well-differentiated HCC. Our results also suggest that GPC3 may be considered as an early marker of liver carcinogenesis because it is able to identify some cirrhotic macronodules with malignant potential.     

p53 immunoreactivity in hepatocellular adenoma, focal nodular hyperplasia, cirrhosis and hepatocellular carcinoma

The prolonged half-life of mutant p53 makes feasible its immunocytochemical detection. In order to assess the pathogenetic role of mutant p53 in regenerative and neoplastc liver disease we studied its immunohistochemical expression in cases of hepatic cirrhosis, hepatocellular carcinoma (HCC), cirrhosis with areas of HCC, hepatocellular adenoma and focal nodular hyperplasia. The study included needle and wedge biopsies of 50 cirrhotic livers, 59 HCCs (36 of them with associated cirrhosis), six adenomas and two focal nodular hyperplasias. Sixty-five HCC fineneedle cytology specimens were also included in the study. There was no immunohistochemical evidence of mutant p53 expression in any of the cases of cirrhotic liver (except for one instance associated with HCC) adenoma or focal nodular hyperplasia. In contrast p53 was detected in 8.5% of HCC cases in the biopsy series and 24% of HCC cases in the fine needle aspiration series. In addition, mutant p53 expression in HCC was positively correlated with tumour grade. According to grade, the distribution of p53 positive immunoreactivity among HCCs was as follows: Grade I-II, 0% of cases in the biopsy series and 9% in the fine needle aspirates; Grade III, 18% in the biopsy series and 55% in the fine needle aspirates; and Grade IV, 40% in the biopsy series. Therefore, mutant p53 expression does not seem to be associated with benign liver lesions but seems to correlate with the progression of HCC through various grades of increasing malignancy.     

Arterial elements and perisinusoidal cells in borderline hepatocellular nodules and small hepatocellular carcinomas

Borderline hepatocellular nodule in the human cirrhotic liver is considered a preneoplastic lesion of hepatocellular carcinoma (HCC). However, the angiogenetic process and changes in perisinusoidal cells (fat-storing cells or Ito cells) during the borderline nodule-HCC sequence have not been investigated. We have investigated intraparenchymal arterial elements and perisinusoidal cells in normal livers, chronic hepatitis, borderline nodules and small HCC, using an immunohistochemical staining for α-smooth muscle actin. In normal livers, chronic hepatitis, cirrhotic nodules and large regenerative nodules, no or few arterial elements were present in the parenchyma, and α-smooth muscle actin-positive perisinusoidal cells were not increased. In borderline nodules, however, there were many intranodular arterial elements, and perisinusoidal cells were significantly increased. In small HCC, there were much more arterial elements, and perisinusoidal cells were increased further. These data suggest that angiogenesis first occurs in borderline hepatocellular nodules and it gradually proceeds during the nodule to HCC sequence along with an increase in perisinusoidal cells. The demonstration of arterial elements and perisinusoidal cells may be useful for the differential diagnosis of large regenerative nodule, borderline hepatocellular nodule and small HCC.     

Immunoreactivity for LN2 and LN3 distinguishes small cell carcinomas from non-small cell carcinomas in the lung

Immunoreactivity to LN2 and LN3, monoclonal antibodies that recognize components of the class II major histocompatibility complex, was assessed in 72 cases of non-small cell lung carcinoma (NSCLC) (32 biopsy specimens, 40 resection specimens) and 64 cases of small cell carcinoma (56 biopsy specimens, 8 resections) of the lung. All cases were reviewed independently by three pathologists for histological classification. Only 1 of the 64 small cell carcinomas showed immunoreactivity for LN2, and none of the 64 cases showed reactivity for LN3. Among the non-small cell carcinomas, 25 of 48 cases were positive for LN2 and 43 of 71 were positive for LN3; the sensitivity was greater for adenocarcinoma (78.5%) than for squamous cell carcinoma (37%). A combined sensitivity of 64.7% was observed when the results of LN2 and LN3 were combined, and this sensitivity was not significantly diminished in the biopsy subset of cases (59.4%). Differentiation within histological subtypes of NSCLC (ie, well, moderate, or poorly differentiated) did not alter test sensitivity. In conclusion, LN2 and LN3, used alone or in combination, appear highly specific for non-small cell carcinoma and moderately sensitive in both biopsy and resection specimens; therefore, these antibodies may be diagnostically useful in distinguishing small cell from non-small cell carcinoma of the lung.     

Synchronous hepatocellular carcinoma and cholangiocarcinoma arising in two different dysplastic nodules.

We present the first reported case of explant cirrhotic liver that had synchronous cholangiocarcinoma and hepatocellular carcinoma arising in two different high-grade dysplastic nodules. The patient was a 55-year-old woman who had hepatitis B virus-associated liver cirrhosis for 3 years. The moderately differentiated cholangiocarcinoma occurred in high-grade dysplastic nodule with a 1.7-fold cell density compared with that of cirrhotic nodule. The hepatocellular carcinoma arose in a nodule-in-nodule pattern within a peripherally low-grade and centrally high-grade dysplastic nodule and had a 2.7-fold cell density compared with that of cirrhotic nodule. By immunohistochemistry, the tumor cells of the cholangiocarcinoma as well as bile ductular cells in dysplastic nodule were diffusely positive for cytokeratin 7, whereas hepatocellular carcinoma cells and dysplastic hepatocytes were negative for cytokeratin 7. The c-kit-positive hepatic progenitor cells were singly scattered between hepatocytes, and their number was highest in cirrhotic nodule and decreased in dysplastic nodule, whereas they were absent in cholangiocarcinoma and hepatocellular carcinoma arising in dysplastic nodules. Proliferation indices were progressively increased in cirrhotic nodule, dysplastic nodule, and cholangiocarcinoma or hepatocellular carcinoma, sequentially. These observations indicate that cholangiocarcinoma as well as hepatocellular carcinoma can develop in dysplastic nodule and that hepatic progenitor cells might play a role in the early stage of cholangiocarcinogenesis and hepatocarcinogenesis.     

Expression of transforming growth factor-beta1 and transforming growth factor-beta receptors in hepatocellular carcinoma and dysplastic nodules.

In this study we analyzed by immunohistochemistry the expression of TGF-beta1 protein and TGF-beta receptors I and II in 4 low-grade dysplastic nodules, 2 high-grade dysplastic nodules, 6 early, 22 small, and 62 advanced hepatocellular carcinomas. The expression of TGF-beta1 protein by hepatocytes was decreased in advanced hepatocellular carcinoma compared with small or early hepatocellular carcinoma(P < .05). Frequent and intense staining of TGF-beta1 protein was noted in the sinusoidal endothelium of advanced hepatocellular carcinomas despite of its decreased staining in hepatocellular carcinoma cells. Reduced expression of TGF-beta receptors I and II compared with surrounding nontumorous tissue were noted from the early hepatocellular carcinoma stage suggesting that down-regulation of TGF-beta receptors is correlated with progression from premalignant to malignant phenotype. Reduced expression of both TGF-beta1 and TGF-beta receptor II in neoplastic hepatocytes were also significantly correlated with increased tumor size and increased proliferative activity(P < .05). These findings suggest that during hepatocarcinogenesis, the inhibitory effects of TGF-beta1 protein on hepatocellular carcinoma cells is outweighed by its effects on stromal elements, which, overall, contributes indirectly to a tumor growth stimulatory environment. Also, the growth-inhibitory effects of TGF-beta1 may have been further negated by reduced TGF-beta receptors on hepatocellular carcinoma cells.     

Differences in growth of transplants of liver, liver hyperplastic nodules, and hepatocellular carcinomas in the mammary fat pad.

Transplantation of fragments of normal rat liver autologously and isologously into the inguinal mammary fat pad permitted survival for up to 75% of grafts for 38 weeks, the longest interval studied. Similarly transplanted hepatocarcinomas grew rapidly and progressively in this site. Neither autologous or isologous transplants of liver hyperplastic nodules displayed obvious growth, although like normal liver, they also persisted for up to 38 weeks. Some persisting hyperplastic cells retained certain characteristic features, but others appeared to revert to a normal morphology. Thus, there is a stage in which hyperplastic cells do not possess the progressive growth ability of neoplastic cells and appear to be capable of reversion to a normal phenotype.