肝细胞癌微循环模式的形态及分布

目的:探讨肝细胞癌中微循环模式的形态结构与分布特点.方法:采用CD34-PAS双重染色法对87例肝细胞癌标本进行染色,判定各种微循环模式组织学结构特点并量化各种微循环模式的分布.结果:肝细胞癌中存在CD34-PAS染色双阳性的内皮细胞依赖性血管,其是血液供应的主要模式;35.63％的病例存在血管生成拟态,且出现肿瘤细胞型和细胞外基质型2种组织学类型;32.18％的病例组织中出现马赛克血管;肿瘤细胞型血管生成拟态、细胞外基质型血管生成拟态和马赛克血管的供血面积之和占全部的36.22％.结论:肝细胞癌组织至少存在4种微循环模式且具有异质性.     

人原发性肝癌间质胶原及其mRNA的表达

用ＡＢＣ方法对２９例人肝细胞癌和２例胆管细胞癌组织及癌旁肝细胞Ｉ型胶原、Ⅲ型前胶原进行免疫组化定位。结果显示两种胶原在细胞外分布是一致的。作者将胶原在肝细胞癌中的分布类型分为两大类：血管样分布和非血管样分布。癌组织内胶原呈血管样分布者，肿瘤易浸润生长及门静脉转移，其肝内转移灶发生率也显著高于非血管分布者，预后较差，推测胶原的分布形式可作为判断肝癌患者预后的一个指标，我们还用ｃＤＮＡ－ｍＲＮＡ原位分     

微环境与肝癌发病机制的研究进展

肝细胞癌是一种在微环境刺激因素作用下由肝细胞基因组积累的突变所导致的常见原发性肝癌。肿瘤微环境是一个动态系统,由肿瘤细胞、间质组织、细胞外基质、组织低氧及饮食、胃肠道微生物群落和微泡等组成,肝脏微环境在肝细胞癌发生,发展和治疗中起重要作用,研究肝细胞癌微环境可为发现肿瘤生成机制和肝细胞癌治疗新靶点提供新的借鉴。     

涎腺腺泡细胞癌临床病理分析并文献复习

目的：探讨涎腺腺泡细胞癌的临床病理学特点及诊断要点。方法：对1例涎腺腺泡细胞癌进行临床资料、病理形态学及免疫组织化学观察,并结合文献对其诊断及鉴别诊断进行探讨。结果：镜下瘤细胞胞体宽大,胞浆嗜碱性呈细颗粒状,核圆形瘤细胞生长呈腺泡状或实性片状生长,部分区域呈乳头状改变,免疫组化显示AE1/AE3(+)、CK8/18(+)、CK7(+)、AAT(+)、S-100(+)。结论：涎腺腺泡细胞癌发病率低,但根据其常见的发病部位及特征性的组织形态,结合免疫组织化学方法,有助于其诊断及鉴别诊断。     

乳腺原发性腺泡细胞癌临床病理诊断

目的探讨乳腺原发性腺泡细胞癌的临床病理特征及诊断和鉴别诊断。方法对1例乳腺原发性腺泡细胞癌进行光镜观察及免疫组化标记。结果癌细胞由同种细胞组成,排列成实性巢状,细胞核圆形,胞质内富含颗粒,胞质嗜酸性或空泡状,透明细胞常见;免疫组化特征性表达CK、lysozyme、α-anti-chymotrypsin等抗体,有助于与其他肿瘤鉴别。结论乳腺腺泡细胞癌非常罕见,需要与乳腺浸润性导管癌、腺肌上皮瘤、颗粒细胞癌等鉴别。     

肝细胞癌和肝硬变中bcl-2蛋白表达与细胞凋亡的关系

目的：研究肝细胞癌和肝硬变中ｂｃｌ－２蛋白表达与细胞凋亡的关系。方法：应用原位脱氧核糖核酸末端转移酶标记法和Ｓ－Ｐ免疫组化技术检测２８例肝硬变和３５例肝细胞癌组织中凋亡细胞的分布、密度及ｂｃｌ－２蛋白的表达。结果：肝细胞癌组织中凋亡细胞密度显著低于肝硬变，且随其恶性程度的增高呈逐渐降低趋势；凋亡细胞在肝硬变中多分布于假小叶周边区域。ｂｃｌ－２蛋白在肝细胞癌组织中的表达强度明显高于肝硬变，但表达的阳性率差异不明显。结论：ｂｃｌ－２通过其表达产物调控肝硬变和肝细胞癌中的细胞凋亡，在肝细胞癌发生中起重要作用，但ｂｃｌ－２蛋白并非细胞凋亡的唯一调控因素     

体外研究整合素αv、α4、β1、β3在小鼠胚泡着床中的作用

整合素(Integrin)是一类存在于细胞表面的跨膜糖蛋白，属于细胞粘附分子(cell adhesion molecules，CAMS)的一大家族，是细胞外基质(ECM)的受体，以受体一配体相对应的形式调节细胞与细胞、细胞与细胞外基质的相互作用，介导细胞的增殖、分化、粘附、迁移等过程。胚泡着床是个复杂的生理过程，包括胚泡孵出、胚泡与子宫内膜的识别、粘附及滋养层细胞向子宫内膜侵入等，     

组织芯片检测星形胶质细胞上调基因1与原发性肝细胞癌病理因素的相关性

背景：星形胶质细胞上调基因1是肿瘤发生转移早期的重要标记分子之一,很有可能成为肝癌临床治疗的分子靶标。 目的：探索原发性肝细胞癌患者星形胶质细胞上调基因1在其癌组织中的表达水平,并推测其在原发性肝细胞癌患者微创疗效中的地位。 方法：采用组织芯片方法和免疫组织化学染色方法检测120例原发性肝细胞癌患者肿瘤组织中星形胶质细胞上调基因1蛋白表达水平,并与其临床病理资料进行相关性分析。 结果与结论：原发性肝细胞癌患者星形胶质细胞上调基因1蛋白表达水平与其临床分期和组织分化程度密切相关,且其星形胶质细胞上调基因1蛋白高表达水平与Ki-67阳性率和肿瘤微血管密度有关,但该蛋白高表达与患者的性别及年龄无相关性。说明星形胶质细胞上调基因1蛋白表达水平与常规恶性检测指标相关,检测该蛋白表达水平对于判断原发性肝细胞癌具有重要意义。     

小鼠卵丘透明质酸结合蛋白的分布

目的：观察小鼠卵丘细胞外基质透明质酸结合蛋白（HABPs）的分布,探讨HABPs对小鼠卵丘的结构和功能的影响。方法：应用免疫荧光双重染色检测HABPs在野生型和bikunin基因敲除小鼠卵丘细胞外基质中的表达。结果：HABPs在两型小鼠卵丘细胞外基质呈阳性表达,HABPs的缺失损害小鼠卵丘细胞外基质完整。结论：HABPs对卵丘细胞外基质的完整是必需的。     

粘附分子与系统性红斑狼疮

粘附分子（adhesion molecules，AM）是指介导细胞与细胞间或细胞与基质间相互接触和结合的一类分子，大都为糖蛋白，分布于细胞表面或细胞外基质（extracellular matrix，ECM）中。粘附分子以配体受体相对应的形式发挥作用，导致细胞与细胞间、细胞与基质间或细胞-基质-细胞之间的粘附。     

Localization of hepatitis B surface antigen in hepatocellular carcinoma

The author respectively examined the localization of hepatitis B surface antigen (HBsAg) in 150 autopsy cases of hepatocellular carcinoma (HCC) with particular attention to growth pattern of HCC. Varying numbers of orcein-positive cells were observed in 42 cases (28.0%) with various distribution patterns, and all the 42 cases were confirmed with immunoperoxidase technique for HBsAg. HBsAg-positive cells were detected in HCC tissue in 10 cases (6.6%) among the 150 cases. Regarding the growth pattern of HCC in the 10 cases, many HBsAg-positive cells were seen in HCCs of the sinusoidal and replacing types, in which the hepatocytes were frequently retained in the cancerous tissue, particularly around the tumor-nontumor boundary. On the other hand, there was no HBsAg-positive cell in the encapsulated type HCC which was the most common (approximately 50%) in the present study and in which the retained hepatocytes were hardly seen. Meticulous histological observation after decoloration of the positive reaction to HBsAg in the sections treated with immunoperoxidase technique and subsequent eosin stain disclosed that HBsAg-positive cells in HCC tissue were retained hepatocytes in 9 cases, and the possibility of HCC cell was not denied in one case. Moreover, HBsAg-positive cells were never detected in tumor thrombi of the portal vein branches and pulmonary metastases.     

GPC-3联合CK-19免疫组化检测在肝细胞癌诊断及鉴别诊断中的应用

目的 研究磷脂酰肌醇蛋白多糖-3（GPC-3）联合细胞角蛋白-19（CK-19）免疫组化检测在肝细胞癌诊断及鉴别诊断中的应用价值。方法 选择2017年1月~2019年1月我院通过肝穿刺或外科手术获取的肝癌标本91例。所有标本按照GPC-3及CK-19检测流程处理,比较肝细胞癌（HCC）、肝内胆管细胞癌（ICC）及肝细胞癌合并肝内胆管细胞癌（CHC）标本中GPC-3及CK-19检测阳性率,及不同分化程度HCC的GPC-3及CK-19阳性率。结果 HCC与CHC中GPC-3阳性率高于ICC,ICC与CHC中CK-19阳性率高于HCC,差异有统计学意义（P＜0.05）;中、低分化的HCC标本中GPC-3、CK-19阳性率高于高分化HCC,差异有统计学意义（P＜0.05）。结论 在肝细胞癌诊断及鉴别诊断中采取GPC-3联合CK-19免疫组化检测,可有效检出HCC,并可与CHC及ICC进行区分,在早期诊断中也具有一定应用价值。     

Hepatitis B virus X mutations occurring naturally associated with clinical severity of liver disease among Korean patients with chronic genotype C infection

Few reports have detailed mutation frequencies and mutation patterns in the entire X region according to clinical status. The aims of this study were to elucidate the relationships between mutation patterns and their frequencies in the X region and clinical status in a Korean cohort and determine specific X mutation types, related closely with liver disease progression. All X mutations were determined by direct sequencing in 184 patients with different clinical features. Mutation rates in the X region in patients with more severe liver disease, hepatocellular carcinoma (HCC) (3.6%) or liver cirrhosis (4%) were always significantly higher than in patients with corresponding less severe forms, chronic hepatitis (2.9%) or asymptomatic carriers (2.1%), but no significant difference in mutation rates was found in terms of HBeAg serostatus. All five mutation types (V5M/L, P38S, H94Y, I127T/N, and K130M and V131I) affecting the six codons were found to be related significantly to clinical severity. Among these, two mutation types (V5M/L and K130M and V131I) were observed more frequently in HBeAg negative patients than in HBeAg positive patients. In conclusion, the results suggest that an accumulation of mutations in the X region contributes to disease progression in chronic patients, at least Korean patients with genotype C. Specific mutation types appears to be related more to severe liver diseases such as HCC or liver cirrhosis. In particular, a novel mutation type (V5M/L) discovered firstly during the present study was found to be associated significantly with HCC.     

Distribution of albumin- and/or alpha-fetoprotein-positive cells in hepatocellular carcinoma

Albumin (ALB)-positive cells were identified by an immunoperoxidase technique in 52 of 53 autopsy cases and in all of 13 surgical cases of hepatocellular carcinoma (HCC). The distribution pattern of ALB-positive cells could be classified into three groups: diffuse, localized, and sparse. The diffuse type was the most common pattern and was usually seen in well or moderately differentiated HCC, showing a trabecular growth pattern. The localized or sparse patterns were more frequent in poorly differentiated HCC showing a compact growth pattern. alpha-Fetoprotein (AFP)-positive cells were detected in 37 of the 53 autopsy cases of HCC and 11 of the 13 surgical cases. The number of AFP-positive cells and the intensity of the immunoperoxidase reaction were roughly proportional to serum AFP levels in most cases. In most regions of HCC, there seemed to be an inverse relationship between the number of ALB- and AFP-positive cells, suggesting tht most HCC cells synthesized only one of the two antigens studied. ALB-positive hepatocytes were found in all of the normal or cirrhotic livers examined and in the tumor-free regions of the HCC-containing livers. In contrast, AFP was not detected in nonneoplastic hepatocytes.     

Glypican-3 as a potential differential diagnosis marker for hepatocellular carcinoma: a tissue microarray-based study

The differential diagnosis between hepatocellular carcinoma (HCC) and benign hepatic lesions is still difficult and new biochemical markers for HCC are required. The aim of this study was to assess the differential diagnostic value of glypican-3 (GPC3) immunostaining in HCC patients. 147 cases of surgically excised HCC tissues, 94 cases from needle biopsies, and tissue microarrays were used for this study. The tissue microarrays contained 449 specimens including: 115 HCC, 25 intrahepatic cholangiocellular carcinoma, 29 lung adenocarcinoma, 23 squamous cell lung carcinoma, 53 ovary adenocarcinoma, 44 renal cell carcinoma, 30 prostate acinar adenocarcinoma, 42 breast carcinoma, 41 gastric carcinoma and 47 colorectal carcinoma. The immunolocalization of GPC3 was measured using immunohistochemical staining. Among 147 surgically excised HCC samples, 87.1% (128/147) were GPC3 positive. No GPC3 expression, however, was observed in paracarcinomatous and cirrhotic tissues. In needle biopsy tissues, GPC3 was positively expressed in 81.9% (77/94). Among tissue microassays, HCCs showed positive GPC3 expression in 55.7% (64/115), while 9.6% (5/52) of lung carcinoma and 5.7% (3/53) of ovary adenocarcinoma also were positively stained. The other tumor types showed negative GPC3 expression. In conclusion, our results show that GPC3 is specifically overexpressed in HCC tissue and may be regarded as a potential marker for differential diagnostic hepatocellular carcinoma.     

Patterns of hepatitis B surface antigen. Localization in cells of hepatocellular carcinoma.

Hepatitis B surface antigen (HBsAg) was identified with immunofluorescence, immunoperoxidase, and aldehyde fuchsin stains within tumor cells in three cases of hepatocellular carcinoma (HCC) from a series of liver biopsies from 172 consecutive cases of HCC. Two patterns of distribution and staining of HBsAg in cells of HCC were observed. In two of the three biopsy specimens, HBsAg was confined to solitary or small groups of tumor cells where a heavily stained inclusion occupied the entire cytoplasm displacing the nucleus. These inclusions corresponded to ground-glass cytoplasm with hematoxylin-eosin. The pattern is different in the other specimen where all the HCC cells in one area of the tumor showed a diffuse peripheral or perinuclear staining of the cytoplasm. In hematoxylin-eosin sections, these tumor cells showed partial transformation of the cytoplasm into the ground-glass appearance.     

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.     

Morphological changes and molecular expressions of hepatocellular carcinoma cells in three-dimensional culture model

Metastatic processes of hepatocellular carcinoma (HCC) are highly associated with the breakdown of extracellular matrix (ECM). However, the regular two-dimensional (2D) culture system, in which only little ECM is involved, fails to provide a well-defined microenvironment for HCC functional research. HAb18G/CD147, a HCC-associated antigen, plays important roles in HCC progression, migration and invasion. In this study, we investigated whether HAb18G/CD147 enhanced the HCC migration and invasion in three-dimensional (3D) culture model through affecting the key molecules and enzymes involved in the metastatic processes, such as focal adhesion kinase (FAK), matrix metalloproteinases (MMPs) and cytoskeleton proteins. We found that, compared with those in 2D cell culture model, the expression of HAb18G/CD147 was significantly increased in 3D cell culture model, together with a high production of MMPs (P < 0.01), an enhanced expression and activation of FAK (P < 0.01) and a changed distribution of F-actin. In addition, the expressions of paxillin and E-cadherin, which enhance the adhesion and migration potentials, were also significantly increased in 3D cell culture model (P < 0.01). All the results suggest that the enhanced expressions of HAb18G/CD147, MMPs, paxillin and FAK changed the distributions of cytoskeleton in the 3D reconstituted basement membrane (BM) and increased the adhesion and invasion potentials of HCC cells.     

大鼠肝癌细胞与I型胶原的黏附特性及其与细胞周期的关系

肿瘤细胞和胞外基质之间的黏附特性与肿瘤的侵袭转移有密切关系。作者从细胞周期的角度,采用微管吸吮技术和细胞同步技术研究了不同细胞周期肝癌细胞与I型胶原裱衬表面的黏附力学特性。结果表明:胸腺嘧啶脱氧核苷、秋水仙碱顺序阻断和胸腺嘧啶脱氧核苷双阻断后释放培养的方法可分别获得G 1期和S期肝癌细胞,平均同步率分别为74.09%和90.39%;在研究剂量和时间范围内,肝癌细胞与I型胶原的黏附力具有浓度和时间依赖性;S期肝癌细胞和I型胶原的黏附力值与G 1期和未同步组(对照组)相应值比较明显降低。结果提示:肝癌细胞经血道转移的侵蚀细胞间质阶段,G 1期细胞可能起更重要的作用。这一研究对全面认识肝癌的转移机理有重要意义。