A combination method for preparing casting and transparent liver specimen

With the development of hepatic surgery and radiology, a more accurate understanding of intrahepatic vessels and hepatic segments is necessary. Previously, research in these fields was primarily by means of dissecting livers, preparing corrosion cast specimens, reconstructing three‐dimensional MSCT images of intrahepatic vessels, and so on. The aim of this study was to search for a new specimen preparing method, which could demonstrate intrahepatic vessels and the relationships between internal vessels and external structures of liver simultaneously. Rabbit livers were prepared with three different techniques in this study: (a) corrosion‐cast technique; (b) transparency technique; and (c) combination technique of cast and transparency. The results showed that the combination of casting and transparent liver specimens in group C could demonstrate both the intrahepatic vessels and the external structures clearly. The relationships between the internal vessels and external structures could also be observed clearly. In conclusion, the combination method for preparing casting and transparent liver specimen created a new approach for the research of intrahepatic vessels, especially for applied basic research of hepatic segments. Clin. Anat. 23:559–562, 2010. © 2010 Wiley‐Liss, Inc.     

原位及游离肝脏可视化数据集获取技术研究

目的建立可供计算机自动识别与三维重建的高精度人体原位肝脏和游离肝脏可视化数据集。方法采用管道灌注法对肝脏进行灌注填充，经数控机床逐层铣切，完成断面图像数码摄影。结果获取无缺损的连续薄层横、冠、矢状断面可视化肝脏图像数据集。结论经灌注后铣切获取的薄层肝脏断面图像能够更好地展示肝内管道系统的断面解剖学数据，有利于计算机准确而快捷地识别与完成肝内管道系统的三维重建。     

可视化肝脏肝内管道灌注及识别

目的：建立计算机可自动识别与三维重建的高精度人体肝内管道可视化数据集。方法：采用多管道搭配灌注法对肝内管道进行分色灌注填充,选择收缩率小、切割特性好、非水溶性的塑料填充剂并分别配以颜色值差异显著的油画颜料,对经过数控机床逐层铣切所获得的肝脏数据集进行计算机自动识别和三维重建。结果：用多管道分色灌注法得到的肝脏断层数据集,完成可视化肝脏的三维重建。结论：多管道分色灌注法能够较好地展示肝内管道系统的断面解剖学数据,有利于计算机准确而快捷地识别与完成肝内管道系统的三维重建,真实反映肝内结构的解剖学特点及管道间的空间毗邻关系。     

Embryology of Extra‐ and Intrahepatic Bile Ducts,the Ductal plate

In the human embryo, the first anlage of the bile ducts and the liver is the hepatic diverticulum or liver bud. For up to 8 weeks of gestation, the extrahepatic biliary tree develops through lengthening of the caudal part of the hepatic diverticulum. This structure is patent from the beginning and remains patent and in continuity with the developing liver at all stages. The hepatic duct (ductus hepaticus) develops from the cranial part (pars hepatica) of the hepatic diverticulum. The distal portions of the right and left hepatic ducts develop from the extrahepatic ducts and are clearly defined tubular structures by 12 weeks of gestation. The proximal portions of the main hilar ducts derive from the first intrahepatic ductal plates. The extrahepatic bile ducts and the developing intrahepatic biliary tree maintain luminal continuity from the very start of organogenesis throughout further development, contradicting a previous study in the mouse suggesting that the extrahepatic bile duct system develops independently from the intrahepatic biliary tree and that the systems are initially discontinuous but join up later. The normal development of intrahepatic bile ducts requires finely timed and precisely tuned epithelial–mesenchymal interactions, which proceed from the hilum of the liver toward its periphery along the branches of the developing portal vein. Lack of remodeling of the ductal plate results in the persistence of an excess of embryonic bile duct structures remaining in their primitive ductal plate configuration. This abnormality has been termed the ductal plate malformation. Anat Rec, 291:628–635, 2008. © 2008 Wiley‐Liss, Inc.     

肝内胆管结石的外科治疗

肝内胆管结石是我国常见病,但其病变复杂,临床治疗具有一定难度。目前肝内胆管结石的治疗原则为“解除梗阻、清除病灶、取净结石、通畅引流”,治疗肝内胆管结石最有效的方法是外科手术。现代医学的发展使肝内胆管结石的外科治疗手段多样化,微创外科治疗理念深入人心。本文主要就近年肝内胆管结石的外科治疗研究进行综述。     

Convoluted bile ducts in the liver of the larval lamprey, Petromyzon marinus L

Summary The three-dimensional structure of the bile ducts and their relationship to the blood vessels were studied in the larval lamprey by scanning electron microscopy of the intact tissue and of biliary and vascular casts. The intrahepatic gall bladder is situated in the cephalic portion of the liver and a cystic duct is connected to a straight intrahepatic common bile duct, which extends to the extrahepatic bile duct at the caudal end of the liver. Several smaller intrahepatic common bile ducts are connected directly to the intrahepatic common bile duct, are convoluted or serpiginous and are surrounded intimately by sinusoids. This arrangement enables the bile ducts to have increased surface area exposed to blood vessels. The functional significance of this arrangement is discussed with respect to the modification of bile through the transport of solutes and the similarity of this bilio-vascular relationship to the peribiliary vascular plexus of the mammalian liver.     

Scanning electron microscopy of mouse intrahepatic structures.

Livers of normal mice were prepared for scanning electron microscopic (SEM) study by fracturing or slicing lobes fixed in situ by perfusion with paraformaldehyde. Fracturing fixed liver exposes surfaces of hepatocytes and sinusoidal endothelial cells, whereas slicing the tissue reveals the internal structures of portal tracts. Earlier studies have delineated the major surface characteristics of hepatocytes and sinusoidal endothelial cells of rats. Surfaces of hepatocytes in the mouse differ from those in the rat by having larger and more numerous peg and hole complexes on the flat intercellular surface and less dense populations of perisinusoidal microvilli. Sinusoidal endothelial cells in the mouse have fewer large fenestrations than do similar cells in the rat; clusters of small fenestrations appear similarly distributed in both species. The surfaces of capsular mesothelial cells, Kupffer cells, bile duct epithelial cells, and endothelial cells of major vessels are similar in rat and mouse.The methods described for preparing liver for SEM examination are simple, rapid, and reproducible. The SEM is a useful tool with which to study intrahepatic surface structures, and its use may allow further correlations to be made between hepatic structure and function in both health and disease.     

Segmentation and reconstruction of hepatic veins and intrahepatic portal vein based on the coronal sectional anatomic dataset

Three-dimensional (3D) reconstruction of intrahepatic vessels is very useful in visualizing the complex anatomy of hepatic veins and intrahepatic portal vein. It also provides a 3D anatomic basis for diagnostic imaging and surgical operation on the liver. In the present study, we built a 3D digitized model of hepatic veins and intrahepatic portal vein based on the coronal sectional anatomic dataset of the liver. The dataset was obtained using the digital freezing milling technique. The pre-reconstructed structures were identified and extracted, and then were segmented by the method of manual intervention. The digitized model of hepatic veins and intrahepatic portal vein was established using 3D medical visualization software. This model facilitated a continuous and dynamic displaying of the hepatic veins and intrahepatic portal vein at different orientations, which demonstrated the complicated relationship of adjacent hepatic veins and intrahepatic portal vein realistically in the 3D space. This study indicated that high-quality 2D images, precise data segmentation, and suitable 3D reconstruction methods ensured the reality and accuracy of the digital visualized model of hepatic veins and intrahepatic portal vein.     

Intrahepatic biliary cysts in deer

The gross and histological appearances of intrahepatic cysts in red and roe deer are described. It is suggested that the structures arise as anomalous developments of the biliary tree. Although not of clinical significance, these cysts may pose a problem at meat inspection.     

Abnormalities of intrahepatic bile ducts in extrahepatic biliary atresia

The infantile cholangiopathies are a group of conditions associated with neonatal jaundice, which include extrahepatic biliary atresia, paucity of intra-hepatic bile ducts and disorders associated with persistence of fetal biliary structures, the so-called ductal plate malformations. Although previously regarded as distinct entities, it has recently been suggested that they may represent parts of a disease spectrum in which the principal process is one of bile duct destruction, the morphological manifestations in individual cases being influenced by the stage of intra-uterine development at which such injury occurs and by the site within the biliary system at which there is maximum damage. To further examine this concept, we have studied liver biopsy specimens from 37 neonates with extrahepatic biliary atresia, with particular reference to abnormalities of the intrahepatic bile ducts. Paucity of intrahepatic ducts, defined as a bile duct: portal tract ratio of less than 0.9, was identified in six cases (16.2%). In eight cases (21.6%) we found concentric tubular ductal structures similar to those observed in ductal plate malformations. In one case, both abnormalities could be demonstrated. Our findings support the concept that there is overlap between the various types of infantile cholangiopathy.     

Preferential differentiation of the bile ducts along the portal vein in the development of mouse liver

Summary The development of bile ducts in the mouse liver was studied histochemically, with special reference to their preferential differentiation around the portal vein. Both portal vein and hepatic vein shared a common origin, the omphalomesenteric vein. In the early development of the liver, haematopoietic cells were predominant around both veins. With the progressive development of intrahepatic bile ducts, the following three steps were observed: cluster formation of type I hepatocytes around the portal vein, formation of primitive bile duct structures and basal lamina, then formation of ducts surrounded by connective tissue structures composed of type I and type III collagens and lectin-binding sites, which were predominant around the portal vein compared to the hepatic vein. These results suggest that the deposition of abundant connective tissue structures around the portal vein is a prerequisite for the cell differentiation and basal lamina formation in the bile duct precursors. A possible mechanism of the aggregation of type I hepatocytes around the portein vein is also discussed.     

Phylogenetic analyses of the hepatic architecture in vertebrates

The mammalian liver has a structural and functional unit called the liver lobule, in the periphery of which the portal triad consisting of the portal vein, bile duct and hepatic artery is developed. This type of hepatic architecture is detectable in many other vertebrates, including amphibians and birds, whereas intrahepatic bile ducts run independently of portal vein distribution in actinopterygians such as the salmon and tilapia. It remains to be clarified how the hepatic architectures are phylogenetically developed among vertebrates. The present study morphologically and immunohistochemically analyzed the hepatic structures of various vertebrates, including as many classes and subclasses as possible, with reference to intrahepatic bile duct distribution. The livers of vertebrates belonging to the Agnatha, Chondrichthyes, Amphibia, Aves, Mammalia, and Actinopterygii before Elopomorpha, had the portal triad‐type architecture. The Anguilliformes livers developed both periportal bile ducts and non‐periportal bile ducts. The Otocephala and Euteleostei livers had independent configuration of bile ducts and portal veins. Pancreatic tissues penetrated the liver parenchyma along portal veins in the Euteleostei. The liver of the lungfish, which shares the same origin with amphibians, did not have the portal triad‐type architecture. Teleostei and lungfish livers had ductular development in the liver parenchyma similar to oval cell proliferation in injured mammalian livers. Euteleostei livers had penetration of significant numbers of independent portal veins from their intestines, suggesting that each liver lobe might receive a different blood supply. The hepatic architectures of the portal triad‐type changed to non‐portal triad‐type architecture along the evolution of the Actinopterygii. The hepatic architecture of the lungfish resembles that of the Actinopterygii after Elopomorpha in intrahepatic biliary configuration, which may be an example of convergent evolution.     

彩超对肝内三种常见强回声病灶的鉴别与诊断研究

目的 观察彩色多普勒超声检查对三种不同肝部强回声病灶（胆管结石、肝内钙化灶、肝内胆管积气）的鉴别诊断。方法 对我院2017年9月~2018年3月收治的肝内胆管结石20例和肝内钙化灶40例、肝内胆管积气20例患者的相关资料和超声资料进行分析,比较三种不同肝部强回声病灶的彩超结果。 结果 20例肝内胆管结石患者中,肝左右叶同时发病的有6 例,左叶占30.00%,右叶占40.00%,彩超声像图显示为结石沿着肝内胆管分布,呈簇状,结石通常为多发,表现为斑点状或条索状、团状以及不规则形,超声检查会出现高强回声,后部通常伴有明显的回声影,肝内胆管存在不同程度扩张,和门静脉分支呈现“平行管”特征。40例肝内钙化灶患者中,肝左叶有患者20例,位于肝右叶患者17 例,肝左右叶同时存在患者有3例,彩超检查可以表现为强回声,可见大小不一,形态为团状或斑块状的影像,通常为单发,可分布于肝内的多处,肝内胆管不存在无明显扩张。在20例肝内胆管积气患者中,有6例患者伴有胆道手术病史,彩超声像图可体现出肝内胆管内有强回声出现,体位发生改变时,其形态和位置均会有变化,如发生的强回声位于肝内胆管的前壁处,后方会伴有“彗尾征”图像。结论 彩超可以实现多角度和多切面的扫描检查,结合不同的声像图信息,可以诊断肝内胆管结石、肝内钙化灶、肝内胆管积气,在临床诊断具有非常重要的作用。     

Secondary joining of the bile ducts during the hepatogenesis of the mouse embryo

Summary Development of the bile duct system of the mouse embryo was studied histologically and by an immunofluorescent technique. The hepatic primordium consisted of cranial and caudal portions. In the liver of young embryos, the hepatic cords were present in the presumptive cysticduct epithelium, and the histology of the presumptive cystic duct epithelium near the hilus was similar to that of the hilus epithelium. The results suggest that at least a part of the cystic duct epithelium develops from the cranial diverticulum of the hepatic primordium. Lumen structures were precursors of intrahepatic bile ducts and originated from type I (immature) hepatocytes. The lumina of the lumen structures appeared near the hilus area first, but most were discontinuous with those of the hepatic ducts. With the progress of development, the discontinuous lumen structures became distributed around the portal vein branches in the central part of the liver parenchyma, and gradually connected with each other and also with hepatic ducts. the discontinuous laminin immunofluorescence also appeared in the endodermal cells around the portal vein branches at the younger stages. Therefore, it is conceivable that the intrahepatic bile ducts originate from discrete cell populations of type I hepatocytes around the portal vein branches and subsequently become confluent, but not from the cells of hepatic ducts.     

Pathology of cholangiocarcinoma

Cholangiocarcinomas (CC) are tumors that arise from the epithelial cell of the biliary tract. They represent the second most frequent primitive liver malignancy after hepatocellular carcinoma. Recent epidemiological data show an increase incidence of CC independently of the increased incidence of cirrhosis. According to their location in the biliary tract, we distinguish intrahepatic, hilar (Klastkin tumors) and extrahepatic CC. In literature, confusion exists around hilar CC that are included, according series, to intrahepatic or extrahepatic CC. However, hilar CC share common clinical, morphological and therapeutic features with extrahepatic CC. So, OMS classification of digestive tumors defined two groups of CC: intrahepatic or peripheral CC which develop from small intrahepatic biliary duct beyond the second segmentation, and extrahepatic CC comprising hilar CC and tumors from common hepatic bile duct. In this chapter, we will describe the different gross features and histological characteristic of CC and will detail the major histopronostic criteria of these tumors.     

New monoclonal antibodies reacting with bile ducts: further insights into the pathogenesis of bile ductular proliferation in biliary diseases.

We have produced a range of monoclonal antibodies which stain human intrahepatic bile ducts of different sizes. Amongst 26 monoclonal antibodies produced, five clones reacted specifically with bile ducts of different sizes, of which three have been maintained in culture and their viability following freezing and thawing confirmed. Staining patterns varied between normal adult liver tissue, normal fetal liver tissue and a variety of hepatobiliary diseases. The antibodies provide further evidence of the immunological heterogeneity of the human intrahepatic biliary tree and support the hypothesis that proliferating bile ductules are derived from periseptal hepatocytes. The preparation of the antibodies, their staining reactions in normal adult, normal fetal and a variety of liver diseases are described.     

INTRANUCLEAR INCLUSION BODIES IN LIVER OF CHRONIC HEPATITIS B WITH DELTA INFECTION

Seventy-two patients with HBsAg-positive chronic hepatitis were studied for serum antibody to delta antigen (antl-HD) by radioimmunoassay. Delta antigen (HDAg) in liver of the 72 patients was also studied immunohisto-chemically using peroxidase-labeled anti-HD. In one of the 72 patients, high titers of anti-HD and intrahepatic HDAg were detected. In this patient, HDAg was localized mainly in nuclei and occasionally in the cytoplasm of hepatocytes. By immunoelectron microscopy, HDAg was observed diffusely In some he-patocytic nuclei, but their nucleoli, perichromatln and heterochromatin were devoid of HDAg. Occasionally, inclusion body-like ring-shaped or irregularly shaped aggregates of HDAg were observed in nuclei. At higher magnification, these aggregates consisted of an HDAg-positive amorphous substance and microtubular structures. In their vicinity, scattered HDAg-positive granules were also present. By conventional electron microscopy, similar inclusion bodies composed of a clustered amorphous substance and microtubular structures 10-20 nm in diameter, together with 20-30 nm irregular granules were observed in hepatocyte nuclei, but no core of Dane particles were found. These novel intranuclear structures were not observed in other patients with chronic hepatitis B. It is suggested that they may represent the characteristic ultra-structure associated with delta infection.     

Histologic evaluation of intrahepatic micrometastases in patients treated with or without neoadjuvant chemotherapy for colorectal carcinoma liver metastasis

In the present retrospective study, we tested the hypothesis that neoadjuvant chemotherapy (NAC) as a treatment for patients with colorectal carcinoma liver metastases (CRLM) may reduce intrahepatic micrometastases. The incidence and distribution of intrahepatic micrometastases were determined in specimens resected from 63 patients who underwent hepatectomy for CRLM (21 treated with NAC and 42 without). In addition, the therapeutic efficacy of NAC was evaluated histologically. Intrahepatic micrometastases were defined as microscopic lesions spatially separated from the gross tumor. The distance from these lesions to the border of the hepatic tumor was measured on histological specimens and the density of intrahepatic micrometastases (number of lesions/mm(2)) was determined in regions close to (<1 cm) the gross hepatic tumor. Of the 21 patients treated with NAC, 13 were identified as having a partial response according to the Response Evaluation Criteria in Solid Tumors (RECIST) guidelines; thus, the overall response rate was 62%. Histologic evaluation of the therapeutic efficacy of NAC was significantly associated with tumor response to NAC according to the RECIST guidelines (p=0.048). In all, 260 intrahepatic micrometastases were detected in 39 patients (62%). Intrahepatic micrometastases were less frequently detected in NAC-treated patients than in untreated patients (5/21 [24%] vs. 34/42 [81%], respectively; p<0.001). There were no significant differences in the distance and density of intrahepatic micrometastases between the two groups (p=0.313 and p=0.526, respectively). In conclusion, NAC reduces the incidence of intrahepatic micrometastases in patients with CRLM, but NAC has no significant effect on their distribution when intrahepatic micrometastases are present.     

Intranuclear inclusion bodies in liver of chronic hepatitis B with delta infection

Seventy-two patients with HBsAg-positive chronic hepatitis were studied for serum antibody to delta antigen (anti-HD) by radioimmunoassay. Delta antigen (HDAg) in liver of the 72 patients was also studied immunohistochemically using peroxidase-labeled anti-HD. In one of the 72 patients, high titers of anti-HD and intrahepatic HDAg were detected. In this patient, HDAg was localized mainly in nuclei and occasionally in the cytoplasm of hepatocytes. By immunoelectron microscopy, HDAg was observed diffusely in some hepatocytic nuclei, but their nucleoli, perichromatin and heterochromatin were devoid of HDAg. Occasionally, inclusion body-like ring-shaped or irregularly shaped aggregates of HDAg were observed in nuclei. At higher magnification, these aggregates consisted of an HDAg-positive amorphous substance and microtubular structures. In their vicinity, scattered HDAg-positive granules were also present. By conventional electron microscopy, similar inclusion bodies composed of a clustered amorphous substance and microtubular structures 10-20 nm in diameter, together with 20-30 nm irregular granules were observed in hepatocyte nuclei, but no core of Dane particles were found. These novel intranuclear structures were not observed in other patients with chronic hepatitis B. It is suggested that they may represent the characteristic ultrastructure associated with delta infection.