[Solitary fibrous tumor of the visceral pleura being suspected of chest wall origin: report of a case].

We report a case of a solitary fibrous tumor (SFT) of the pleura which is suspected of chest wall tumor. A 52-year-old female was admitted to our hospital because of epigastralgia and body weight loss. Chest X-ray and computed tomography showed a circumscribed mass of 35 x 22 mm in diameter arising from the parietal pleura. Positron emission tomography showed uptake valve of 1.5. SFT of chest wall origin was suspected and performed video-assisted thoracic surgery. The pedunculated tumor attached to the visceral pleura. The tumor was diagnosed as a benign SFT in intraoperative diagnosis. Long term clinical follow-up is recommended for patients with SFT, because the tumor recurrence and malignant transformation may occur in tumors with benign histological features.

Beta-catenin mutations are frequent in hepatocellular carcinomas but absent in adenomas induced by diethylnitrosamine in B6C3F1 mice.

Activating mutations in the region of the beta-catenin gene corresponding to the NH2-terminal phosphorylation sites of glycogen synthetase kinase 3beta have been causally implicated in carcinogenesis. In this study, the beta-catenin exon 3 was examined in hepatic lesions induced by diethylnitrosamine in B6C3F1 mice. PCR and DNA sequencing detected seven beta-catenin mutations in 13 samples dissected from hepatocellular carcinoma tissues, but none in 14 hepatic adenomas. All of the mutations were found in codon 41 encoding a threonine residue, one of the possible glycogen synthetase kinase-3beta phosphorylation sites. Although beta-catenin protein was immunohistochemically stained mainly on the cell membrane in preneoplastic hepatocytic foci and most adenomas, as observed in normal hepatocytes, it was detected in the cytoplasm and nuclei in addition to the cell membrane, indicating stabilization of the protein in HCCs. This shift in staining was observed not only in tumors with mutations, but also in examples lacking exon 3 mutations. Our data demonstrate that beta-catenin alterations may be important for malignant progression during multistep hepatic carcinogenesis in mice.

A case of apocrine adenocarcinoma associated with hamartomatous apocrine gland hyperplasia of both axillae.

We report an apocrine adenocarcinoma of the left axilla associated with hamartomatous apocrine gland hyperplasia of both axillae. The patient, a 69-year-old man, presented with no symptoms or complaints other than an oval mass felt in the left axilla. The mass was resected and histopathological examination revealed a papillary apocrine adenocarcinoma located within hyperplastic apocrine glands. Because gallium scintigraphy performed after the operation still showed bilateral abnormal uptakes, skin and subcutaneous tissues of the bilateral axillary areas were resected. Histological examination demonstrated marked multilobular hyperplasia of the apocrine glands. These hyperplastic glands did not show distinct atypia, and there was no evidence of tumor remnants in the left axilla. The patient has shown no signs of local recurrence or metastasis at 20 months' follow-up. To our knowledge, this is the first case of malignant transformation of hamartomatous apocrine gland hyperplasia (apocrine gland organic hamartoma or apocrine nevus).

Genetic classification of combined hepatocellular-cholangiocarcinoma.

Combined hepatocellular-cholangiocarcinoma (combined HCC/ CC) is a rare form of liver neoplasms showing both hepatocellular (HCC) and bile duct differentiation (CC). In an attempt to clarify the clonality and genetic/phenotypic relationships in the evolution of these neoplasms, we microdissected multiple HCC and CC foci and studied allelic status of chromosome arms 1p, 1q, 3p, 4q, 5q, 6q, 8p, 9p, 10q, 11q, 13q, 16q, 17p, 17q, 18q, and 22q. Overall, the highest frequency of loss of heterozygosity (LOH) was seen on 4q and 17p, followed by 8p and 16q. Of the 11 cases studied, 3 cases did not show any of the identical allelic losses between HCC and CC foci, indicating the biclonal nature. The remaining 8 cases showed multiple allelic losses shared between both components, strongly suggestive of a single clonal derivation. Moreover, 4 of the 8 cases showed additional or divergent allelic losses at more than 1 chromosomal locus only in HCC and/or CC foci. Thus, this heterogeneity was shown to affect the phenotypic diversity of the tumor. Summarizing the genetic patterns, combined HCC/CC could be classified into the following 3 possibilities: (1) collision tumor in which 2 independent neoplastic clones develop at close proximity; (2) single clonal tumor with homogeneous genetic background in both components--histological diversity is thus a manifestation of divergent differentiation potential of a single clone; (3) single clonal process in which genetic heterogeneity in the process of clonal evolution within the tumor parallels histologic diversity; therefore, the tumor in this category is mainly composed of mosaics of closely related subclones.

Papillary adenoma of the distal common bile duct.

A 73-year-old man with a papillary adenoma located in the distal common bile duct is reported. He underwent pylorus-preserving pancreatoduodenectomy. The lesion in the common bile duct featured papillary proliferation of the epithelium and fibrous elements with diffuse infiltration by inflammatory cells. Positive staining for MIB-1 (Ki-67) and p53 was identified in the nuclei of the proliferative epithelium. These findings suggested the malignant potential of this lesion. Further progress in imaging diagnostic techniques should increase the frequency with which such lesions are discovered. Even now, if mural irregularities and defects are found in the extrahepatic biliary system, especially the distal common bile duct, the possibility of such borderline biliary adenoma should be taken into consideration when making a diagnosis.

Successful treatment of a patient with nephrotic syndrome associated with chronic lymphocytic leukemia.

We report the case of a 62-year-old man with nephrotic syndrome associated with stage B chronic lymphocytic leukemia (CLL). Kappa Bence Jones proteinuria and the glomerular deposition of kappa-light chain were observed. Although treatment with cyclophosphamide and prednisolone tended to reduce the level of proteinuria, the administration of angiotensin-converting enzyme inhibitor, enalapril, resulted in complete remission of nephrotic syndrome.

Protein-loss into retroperitoneal lymphangioma: demonstration by lymphoscintigraphy and blood-pool scintigraphy with Tc-99m-human serum albumin.

A rare, benign congenital lymphangioma has been reported to occur frequently in the neck and axilla, but rarely in the retroperitoneal space. We report a case of a retroperitoneal lymphangioma associated with hypoproteinemia caused by protein-loss into the tumor. In this case, lymphoscintigraphy with subcutaneously injected Tc-99m-human serum albumin (HSA) disclosed the communication between the tumor and the lymphatic system, and sequential abdominal scintigraphy with intravenously injected Tc-99m-HSA revealed the protein loss into the tumor. Abdominal scintigraphy with Tc-99m-HSA injected intravenously or subcutaneously is occasionally useful for determining the etiology of hypoproteinemia.

[Apolipoprotein E immunoreactivities in normal human ovary and ovarian neoplasms].

An immunohistochemical study on localization of apoE in normal human ovary and various ovarian neoplasms was performed using polyclonal anti-human apoE antibody. In the normal tissues, apoE immunoreactivity was found in folliculus (theca interna and stratum granulosum), and most intensely in a corpus luteum. The stroma cells were not stained. Among various ovarian tumours, positive staining was found in granulosa cell tumour, teratoma and dysgerminoma. Close relationship between apoE and steroid synthesis was suggested by the distribution of apoE in the studied tissues. The present results show that an immunohistochemical study for apoE may be helpful for the diagnosis of steroid producing tumours.

[Apolipoprotein E immunoreactivity in normal and abnormal adrenal tissues].

Immunohistochemical study on localization of apo E in normal and abnormal adrenal tissues was performed using polyclonal anti-human apo E antibodies. In normal adrenal glands, the cortex showed intense staining, in particular, in the zona fasciculata and the zona reticularis, while the medulla showed weak staining. In a case of adrenal hyperplasia, apo E immunoreactivity was equivalent to that in the normal cortex. In adenomas (primary aldosteronism, n = 7; Cushing's syndrome, n = 2; others, n = 2), apo E was stained normally or to some extent weakly in comparison with the normal cortex. Adrenocortical carcinoma showed only faint staining. The results indicate that apo E may relate to the functional activity of adrenal cortex, and suggest that immunohistochemical study for apo E may help to distinguish the malignancy from benign adrenal neoplasms.

[Multiple myeloma with a solitary osteosclerotic legion on seventh cervical vertebra].

The osseous manifestation of multiple myeloma is well known as the osteolytic or osteoporotic feature. On the other hand, there are rare cases of general osteosclerotic manifestation as myeloma variants. We report a case of multiple myeloma with solitary osteosclerotic legion in the cervical vertebra. A 60-year-old-man was admitted with paralysis of both arms. The cervical roentogenogram showed the osteosclerotic change of the seventh cervical vertebra. The pathological study of surgical bone biopsy from the vertebra revealed osseous and severe fibrotic change and accumulation of plasma cells in the residual bone marrow. Clusters of plasma cells were also observed in the bone marrow of ileac bone. In addition, IgG-lambda type M protein was seen in the serum. Therefore we diagnosed this case as the osteosclerotic multiple myeloma. We then analyzed the cytokines known to influence bone formation, and found that the bone marrow serum TGF-beta and PDGF levels were increased compared with normal control. These results may suggest that the preferential increase of osteosclerotic cytokines caused the osteosclerotic changes of bone marrow.

[Basaloid-squamous carcinoma of the esophagus: report of a case].

We report herein the case of a 66-year-old man with basaloid squamous cell carcinoma (BSCC) of the esophagus. There are only 60 cases of BSCC of the esophagus previously reported in Japan. In our patient, endoscopic findings revealed a type 2 in the lower intrathoracic esophagus (Lt), and the tumor was intact with the aorta (T2). A biopsy suggested that it was adenosquamous cell carcinoma. An operation was done on June 11, 1996. With open thoracotomy, esophagectomy was performed with mediastinal lymphnode dissection by posterior mediastinal esophagogastrostomy. Histologically, the lesion of the tumor with ulceration was composed of BSCC, and other lesion was composed of typical squamous cell carcinoma (SCC). The immunohistochemical findings of the respected specimens led us to suspect that the basal-layer-type SCC had transformed into BSCC by undergoing differentiation and expansive proliferation. According to the prognosis, eventually the patient died of pneumonia due to methicillin resistant Staphylococcus aureus (MRSA) and candida 4 years and 9 months after the surgery.

Primary small cell carcinoma of lacrimal sac: case report and literature review.

OBJECTIVE: We present the first reported case of primary small cell carcinoma of the lacrimal sac. CASE REPORT: A 67-year-old Japanese woman was referred to our department with a two-month history of left medial canthal swelling, epiphora and occasional nasal bleeding. Nasal endoscopy revealed a readily bleeding tumour in the left inferior meatus. Computed tomography and magnetic resonance imaging scans demonstrated that the tumour was mainly located in the left lacrimal sac. Histopathological studies of a biopsy specimen revealed small cell carcinoma. The patient was treated with four cycles of chemotherapy consisting of cisplatin and etoposide, in combination with radiotherapy. There was no evidence of recurrence or metastasis for five years. CONCLUSION: Small cell carcinoma originating in the head and neck region has been reported to be highly aggressive and to have a poor prognosis. We report a case of primary small cell carcinoma of the lacrimal sac successfully treated with chemo-radiotherapy.

Absence of p53 mutations in methylnitrosourea-induced mammary tumors in rats.

Administration of methylnitrosourea (MNU) to female rats during or before puberty induces a high incidence of mammary tumors, most of which contain a G to A transition at the second base of H-ras codon 12. However, this mutation alone is presumably not sufficient for normal mammary epithelial cells to develop into neoplastic cells, because it can be detected in mammary tissues prior to the appearance of tumors. To clarify whether p53 genetic changes may complement the H-ras mutation in mammary carcinogenesis, we investigated nine MNU-induced mammary tumors of F344 rats by polymerase chain reaction-mediated denaturing gradient gel electrophoresis (DGGE). However, no mutations were identified in p53 exons 5 through 8, which include the known mutational hot spots. Our data thus indicate that p53 mutations are not involved in MNU-induced rat mammary carcinogenesis.

Absence of p53 mutations and various frequencies of Ki-ras exon 1 mutations in rat hepatic tumors induced by different carcinogens.

Mutations of p53 and Ki-ras exon 1 were investigated in rat hepatic lesions induced by four kinds of hepatocarcinogenic protocols: continuous feeding of 3'-methyl-4-dimethylaminoazobenzene (3'-Me-DAB), daily intraperitoneal injection of aflatoxin B1 (AFB1), and the Solt and Farber regimen (Nature 236:701-703, 1976), in which diethylnitrosamine (DEN) or nitrosomethylurea (NMU) was used as initiating agents. DNA from microdissected tissue sections was amplified by the polymerase chain reaction (PCR) directly using primers for p53 exons 5-8 and Ki-ras exon 1 and analyzed for mutations by denaturing gradient gel electrophoresis (DGGE) or constant denaturant gel electrophoresis (CDGE). One or both of the p53 PCR primers were located within introns to prevent amplifying the p53 processed pseudogenes. In a total of 59 hepatocellular carcinomas (HCCs), no p53 aberrations were detected, indicating that p53 mutations are not very important in rat hepatic carcinogenesis. On the other hand, Ki-ras codon 12 mutations were found at low frequency in HCCs, hyperplastic foci, and cholangiofibroses induced by 3'-Me-DAB and by AFB1 but not in the lesions induced by the Solt and Farber regimen. Although Ki-ras codon 12 mutations are generally infrequent in rat hepatic tumors, their incidence thus appears to vary depending on the carcinogen used for their induction.

Intrahepatic transplantation of normal hepatocytes prevents Wilson's disease in Long-Evans cinnamon rats.

BACKGROUND & AIMS: Long-Evans cinnamon (LEC) rats are an animal model of Wilson's disease. This study investigated whether intrahepatic transplantation of normal hepatocytes can prevent Wilson's disease in LEC rats. METHODS: Hepatocytes of newborn Long-Evans agouti (LEA) rats were transplanted into the livers of LEC rats before onset of hepatic disease. Expression of normal transcripts from the gene linked to Wilson's disease was investigated at 30 weeks of age by reverse-transcription polymerase chain reaction and in situ hybridization. RESULTS: Eight of 16 (50.0%) untreated LEC rats died of hepatic failure during 20-30 weeks of age. Of the 27 LEC rats with transplanted cells, 2 (7.4%) died of hepatic failure and 4 died of ileus complicated by the surgical treatment. In the recipient livers, the transplanted cells comprised 4%-20% of the hepatocyte populations, expressing normal messenger RNA transcribed from the Wilson's disease gene, and hepatic copper deposition was reduced to approximately 60% of that in untreated LEC rats. CONCLUSIONS: Transplantation of normal hepatocytes prevents Wilson's disease in LEC rats.

Morphology, proliferation and apoptosis of mouse liver epithelial cells cultured as spheroids.

The MLEC10 is an epithelial cell line derived from an untreated, normal C3H/HeN mouse liver. We previously demonstrated that tumorigenic variants from this cell line produced moderately differentiated hepatocellular carcinomas in nude mice. However, it has remained unclear whether the parental MLEC10 cells represent immortalized hepatocytes or so-called oval cells, both of which may serve as precursors for hepatocellular neoplasms. In this study, we performed 3-dimensional, spheroid culture of the MLEC100 cells in order to facilitate histological assessment of their lineage. Spheroidal aggregates were formalin-fixed and embedded in paraffin for routine light-microscopic observation of hematoxylin and eosin-stained sections. Histopathologically, the MLEC10 cells were indistinguishable from immature hepatocytes and distinct from oval cells. At the electron-microscopic level, their hepatocytic nature was evidenced by bile canaliculus structures and glycogen storage. Intriguingly, the spheroids contained fragmentary material reminiscent of Councilman bodies, implying apoptosis of the hepatocytes. Although the cells significantly proliferated during the first three days of culture, apoptotic death then resulted in a 75 % decrease in viable cell number. Thereafter, both apoptosis and cell division appeared silent, the numbers being unchanged. Expression of the p53 tumor suppressor gene became gradually elevated, correlating positively with growth arrest, but negatively with apoptosis, suggesting that the cell death occurred independently of p53. Our results indicate that at least some liver epithelial cell lines derived from untreated murine livers exhibit a hepatocytic morphology in spheroid culture. Also, the present culture system provides a useful tool for investigating biological phenomena, e.g. apoptosis, specifically involving liver cells, under 3-dimensional conditions.

Hepatocytic cells form bile duct-like structures within a three-dimensional collagen gel matrix.

It has been suggested that hepatocytes have the ability to form bile ductal structures during normal development and in various pathological conditions of the liver. In the present study, we attempted to establish an in vitro model of ductal morphogenesis of hepatocytic cells by combining an aggregate culture and a type I collagen gel culture. When spheroidal aggregates of rat or mouse primary hepatocytes were embedded within the collagen gel matrix and then cultured with a medium containing a fibroblast-conditioned medium, the aggregates extended many dendritic processes composed of a trabecular arrangement of cells. Dendritic morphogenesis was also seen in embedded aggregates of immortal liver epithelia] cell lines, which spontaneously emerged during long-term cultures of mouse primary hepatocytes. A similar morphogenesis was induced by the presence of insulin in the medium. Although epidermal growth factor (EGF) and hepatocyte growth factor (HGF) showed only a small effect on the morphogenesis of most of the hepatocytic cells when used alone, these factors, especially EGF, enhanced the morphogenetic effect of insulin. Electron microscopical observations revealed luminal structures lined by microvilli within these dendritic processes, indicating ductal differentiation. Immunocytochemically, the dendritic processes were positive for cytokeratin 19, a marker for bile duct cells. On the other hand, an H-ras-transformed mouse liver epithelial cell line and rat hepatocellular carcinoma cell lines did not demonstrate the organized morphogenesis. Our results indicate that hepatocytic cells can produce bile duct-like structures in the presence of the type I collagenous matrix and soluble morphogenetic factors.