[A case of advanced gastric cancer responding to chemotherapy and radiotherapy].

A thirty three-year-old male complaining of vomiting was diagnosed as having type 3 advanced gastric cancer of upper stomach and multiple liver metastasis, and had undergone total gastrectomy. The conclusive stage was P2H2n4se stage IVb. Intraoperatively, ethanol injection was performed for the liver metastasis under ultrasonography, and CDDP 100 mg was injected into the intra-abdominal cavity. Postoperative adjuvant therapy was added using oral fluorouracil and OK-432. Then we utilized FP chemotherapy (consisting of 5-FU and cisplatin) and radiotherapy for the bone metastasis. The patient survived 4 years and 4 months with good quality of life.

Superficial spreading-type mucosa-associated lymphoid tissue (MALT) lymphoma in the stomach--a case report.

We presented a case of a 71-year-old woman who presented irregular mucosal folds with tiny depressions and multiple erosions with ulceration in the stomach, on endoscopy and roentgenography. Histological findings from biopsy specimens showed dense and mildly atypical lymphoid-like cells invading the submucosa. Immunohistochemical examinations revealed monoclonal reactivity of Heavy-chain. She was then diagnosed to have low grade malignant lymphoma in the stomach and underwent total gastrectomy. The resected cancer demonstrated the pathological features including small lymphocytic plasmacytoid cells and lymphoepithelial lesions characteristic of a mucosa-associated lymphoid tissue (MALT) lymphoma. The lymphoma cells expressed B cell markers with a phenotype of IgG and lambda type. It is difficult to make a firm diagnosis of a MALT lymphoma from only small biopsy specimens. Accordingly it is necessary to complete the observed clinical process, endoscopic and roentgenographic findings with immunohistochemical characteristics to determine diagnosis. The MALT lymphoma should be treated as a low-grade malignancy and diagnosis sufficiently early can lead to a favorable prognosis.

An alpha-fetoprotein-producing human gallbladder carcinoma cell line (KMG-C) shows more aggressive biologic behavior than an alpha-fetoprotein-non-producing cell line.

We have established a new human alphafetoprotein (AFP)-producing gallbladder carcinoma (GBC) cell line, termed KMG-C, from a 67-year-old Japanese male. KMG-C and its reconstituted tumors in nude mice showed the morphological features of an adenocarcinoma. Functionally, KMG-C secreted AFP, carcinoembryonic antigen, and carbohydrate antigen 19-9, as well as 7 serum proteins, including albumin and C-reactive protein. KMG-C showed more malignant biological behavior than an AFP-nonproducing GBC cell line, KMG-A, established originally from the tumor of the same patient; KMG-C had a shorter doubling time, higher tumorigenicity, and an aneuploid DNA index. Our results suggest that AFP-producing GBC cells may have more malignant biological characteristics than AFP-non-producing GBC cells in GBCs having both of these components.

[Complete response in a case of unresectable gastric cancer treated by combined chemoimmunotherapy of MMC, 5-FU and OK-432].

A 68-year-old female patient with unresectable advanced gastric cancer was treated by combined administration of MMC (10 mg/month), 5-FU (200 mg/day) and OK-432 (5KE/2 weeks). Two months after starting the treatment, there was a diminution in the serum CEA level. Six months later, the CEA had decreased to the normal level, primary and metastatic sites completely disappeared and no cancer cell was confirmed by endoscopic biopsy. The patient has survived for 33 months of a state of complete response.

[A case of advanced gastric cancer with liver and lung metastasis effectively treated by combined chemo-immunotherapy of MMC, 5'-DFUR, OK-432].

A sixty-eight-year-old male patient was diagnosed as having inoperable advanced gastric cancer with liver and lung metastasis. The patient was treated by combined chemo-immunotherapy of MMC 10 mg/M, 5'-DFUR 800 mg/day and OK-432 5 KE/2 W. Six months after commencing chemotherapy, CT-scan and upper GI series revealed that metasized liver tumors and stomach lesion were remarkably decreased in size and no cancer cell was confirmed by endoscopic biopsy. Further, the metastatic lung tumor has disappeared on chest X-ray. The patient had been well without any evidence of tumor re-progression for over one year, but from July the liver tumor began to metastasize again and the patient eventually died of liver metastasis on Jan. 1, 1993.

Anti-KCA-3, a monoclonal antibody reactive with a rat complement C3 receptor, distinguishes Kupffer cells from other macrophages.

A new mAb, designated anti-KCA-3, was developed against rat Kupffer cells. The reactivity of anti-KCA-3 was restricted to macrophages with preferential binding to Kupffer cells; only a few macrophages in the spleen, lymph nodes, lungs, and intestine stained with the antibody. A very small number of peritoneal resident and exudate macrophages reacted with the antibody and no reactivity was seen within the thymus, skin, heart, kidneys, brain, peripheral blood, and bone marrow. KCA-3 was expressed predominantly by the Kupffer cells in the periportal region rather than in the centrilobular region of the hepatic lobules. The cells in the portal tract did not stain with the antibody. The staining of the cytosmears and FACS analysis of the Kupffer cell fraction isolated from hepatic sinusoidal cells by centrifugal elutriation revealed that as many as 62% and 49% of the cells were stained with anti-KCA-3, respectively. Immunoelectron microscopic study of the liver indicated that expression of KCA-3 on Kupffer cells was limited to the plasma membrane facing the sinusoid rather than the space of Disse. Immunoprecipitation and SDS-PAGE analysis demonstrated KCA-3 to have a m.w. of approximately 50 kDa under both reducing and nonreducing conditions. After treatment of KCA-3 with N-glycanase, there was no significant change in the m.w., indicating KCA-3 was not highly glycosylated. C3b- and iC3b-mediated rosette formation between Kupffer cells and sensitized SRBC was inhibited by the antibody, implying that KCA-3 functioned as a complement C3 receptor or complement receptor-associated molecule. Furthermore, KCA-3 was eluted from C3b-Sepharose but not HSA-Sepharose after incubation with Kupffer cell lysate, indicating that KCA-3 directly binds C3b. The cell distribution, ligand-binding specificity, and biochemical properties of the protein were found to be different from the complement C3 receptors previously described. Because OX42 (antibody reactive with the rat CR3 receptor) inhibited complement C3-mediated rosette formation with peritoneal resident macrophages but not with Kupffer cells, the findings suggest that C3-mediated binding to Kupffer cells and to peritoneal macrophages is mediated by two different receptors. We conclude that anti-KCA-3 recognizes a novel type of complement C3 receptor preferentially expressed on Kupffer cells.

Early gastric cancer with juxta lymph node (n3) metastasis.

From 1976 to 1992, a total of 714 cases of early gastric cancer (EGC) were treated by resection in the First Department of Surgery, Kurume University Hospital. For EGC, the overall lymph node metastasis rate is generally 10%, with the lymph node metastasis observed mainly in the Group 1 perigastric nodes in both mucosal cancer and submucosal cancer. Three rare cases of an early gastric cancer with Group 3 juxta regional lymph node metastasis are described because Group 3 lymph nodes are rarely discovered. To our knowledge, only 14 other cases of EGC with juxta regional lymph node metastasis have been reported in the Japanese literature.

Early gastric cancer associated with synchronous multiple liver metastasis--two rare cases.

From 1976 to 1991, 1,640 cases of gastric cancer were resected in our department. Of these, 659 (40.2%) cases were early gastric cancer (EGC). The prognosis after resection of an EGC is generally good, with a 5-year survival rate of 95%. The incidence of an EGC with synchronous liver metastasis is very rare. To our knowledge, only 15 cases of an EGC with synchronous liver metastasis have been reported in the Japanese literature. From 10 cases adequately described the EGC with synchronous liver metastasis derived only from sm-cancers (cancers confined to the submucosa) of medullary type, histologically, more commonly occurring in males than EGC generally, (of elevated type) and with positive lymph node metastasis. Here we report two additional rare cases of EGC with liver metastasis.

Retrospective studies of synchronous double early gastric cancer.

From 1984 to 1990, 922 cases of gastric cancer underwent resection in the First Department of Surgery, at Kurume University Hospital. Of these, 381 (41.3%) cases were early gastric cancer (EGC). Among these EGC, synchronous double early gastric cancer (double EGC), according to the diagnostic criteria of Moertel, was found in 49 (12.9%) cases involving 98 foci. In cases of a double EGC, the primary and secondary foci can be distinguished. Double EGC occurred more frequently in males than females (7.1:1). In the elderly, the incidence of a double EGC was higher than the incidence of a single EGC. Double EGC occurred more frequently in the lower third of the stomach than a single EGC. And a macroscopically elevated and histologically differentiated type primary focus was more frequently found in double EGC than in single EGC. Of the 49 secondary foci in the double EGC, 12 (24.5%) foci were a microcarcinoma of less than 5 mm in greatest diameter, and 48 (98%) were limited to the mucosa (m-cancer). Therefore, a preoperative detailed examination of the whole stomach by endoscopy and an intraoperative careful search under gastrotomy must be made in the remnant of the stomach in cases of distal or proximal partial gastrectomy to ensure adequate excision.

Para-aortic lymph node dissection for the treatment of advanced gastric cancer.

Para-aortic lymph node (No. 16) metastasis has been accepted as a factor responsible for non-curability in gastric cancer surgery. Recently in Japan, after detailed experimental and clinical studies of the lymph flows from the stomach and/or perigastric nodes to these No. 16 nodes, it is now considered to be curable. From 5 autopsy cases, the mean number of No. 16 nodes has been determined to be 43. The frequency of positive No. 16 metastasis (No. 16(+)) was found to be 24%, occurring in 35 of 144 cases with advanced gastric cancer following R3 gastrectomy plus No. 16 dissection. The 5 year-survival rate of R3+ No. 16 dissection was 70.1% in n1(+), 49.8% in n2(+) and 24.3% in n3(+) and had better prognoses (by Kaplan-Meier statistics) than those without dissection of the No. 16 nodes. Only 37 patients in the Japanese literature have had No. 16 dissections, and were confirmed histologically to have had No. 16(+) and subsequently survived more than 5 years. The number of those surviving No. 16(+) can be expected to increase in the near future with the adoption of R3 gastrectomy plus No. 16 dissection.

A new human cholangiocellular carcinoma cell line (KMC-1).

We have recently established a cholangiocellular carcinoma (CCC) cell line, designated KMC-1, from a nude mouse subcutaneous tumor which developed after inoculation of a surgically resected peripheral type CCC from a 62-year-old Japanese male patient. KMC-1 cells grew over a 26-month period and passaged 57 times. These cells retained the morphologic characteristics of both the original tumor and the subcutaneous tumor in the nude mouse, which mainly consisted of irregular tubules and invaded surrounding interstitial tissue in part with an indurate pattern. KMC-1 cells grew in a monolayer pavement-like cell arrangement with tubular formation in part. Some cells and/or glands had a mucin-like substance inside. The doubling time of KMC-1 cells growing in serum-containing medium was 54 h at passage 31. Cell growth in serum-free medium was slow but steady. The number of chromosomes was distributed in range from 73 to 83 with modes of 76 and 78. KMC-1 cells secreted some tumor markers such as DUPAN-2, CA125, TPA, hCG, CA19-9 and ferritin, however, the secretion of DUPAN-2, and CA19-9 and ferritin were only detectable in serum-containing and serum-free medium, respectively. These findings suggest that KMC-1 cells will provide a variety of experimental models for research on CCC and the mechanisms of tumor marker secretion.

Establishment and characterization of a new human extrahepatic bile duct carcinoma cell line (KMBC).

A new human extrahepatic bile duct carcinoma cell line (KMBC) was established from a serially transplanted tumor in nude mice that originated from a surgically resected tumor from a 73-year-old Japanese man; the cell line has been maintained for 5 five years. KMBC cells proliferate in a monolayered sheet with a population doubling time of 30 hours. Chromosome number was distributed in a range from 37 to 44, with modal numbers of 40 and 41. KMBC cells and the reconstituted tumor in a nude mouse showed moderately to poorly differentiated adenocarcinoma and possessed various functional characteristics of extrahepatic bile duct carcinoma. KMBC cells secreted carbohydrate antigen 19-9, tissue polypeptide antigen, carcinoembryonic antigen, ferritin, beta 2-microglobulin, fibronectin, and alpha 2-macroglobulin and produced glutamic oxaloacetic transaminase and alkaline phosphatase. KMBC is the second established cell line that originated from a human extrahepatic bile duct carcinoma in the world literature, and it will be applicable to various experiments.

[Coexistence of multiple metastatic brain tumors from thyroid carcinoma and cerebral aneurysms].

A 73-year-old woman was admitted with coexistence of multiple metastatic brain tumor and cerebral aneurysm. She has been operated on for a tumor of the thyroid gland 3 years before. Computed tomography (CT) scan revealed well-defined, homogeneously enhanced masses in the left frontal and right posterior temporal region. Right carotid angiography showed a tumor stain supplied by the branch of the fronto-polar artery. Left carotid angiography showed an aneurysm at the anterior communicating artery and a tumor stain supplied by the posterior temporal artery. Total removal of the bilateral tumor and aneurysmal neck clipping were performed simultaneously with bi-frontal craniotomy and right temporal craniotomy in April of 1988. Histology of tumor was papillary carcinoma. The surgical indication and treatment for patients with coexistence of multiple brain tumor and cerebral aneurysm were discussed.

Monoclonal antibodies to rat Kupffer cells. Anti-KCA-1 distinguishes Kupffer cells from other macrophages.

Two monoclonal antibodies, anti-KCA-1 and anti-KCA-2, directed against rat Kupffer cells (hepatic sinusoidal macrophages) were developed. Immunohistologic studies of the liver and analysis of isolated hepatic cells by immunofluorescence and flow cytometry showed that the reactivity of these antibodies was restricted to macrophages. Both KCA-1+ and KCA-2+ cells were located predominantly in the periportal region; in contrast, Ia+ sinusoidal cells were located primarily in the centrilobular region. Macrophagelike cells within the portal tracts expressed KCA-2 but not KCA-1. These findings indicate the presence of heterogeneity within the macrophage population of the liver. Anti-KCA-1 reactivity appeared to be almost entirely restricted to Kupffer cells; only a few macrophages in the thymus and a small number of cells in the bone marrow expressed KCA-1. In contrast, KCA-2 was more widely distributed; splenic, lymph node, and intestinal macrophages were intensely stained with anti-KCA-2. These studies indicate that KCA-1 is a marker of Kupffer cells.

A new human pleomorphic hepatocellular carcinoma cell line, KYN-2.

A new human hepatocellular carcinoma (HCC) cell line, KYN-2, has been established from a surgical specimen obtained from a 52-year-old Japanese male HCC patient. The originally resected HCC was classified as pleomorphic HCC corresponding to Edmondson-Steiner's grade III with a thick trabecular to solid arrangement. The cell line has been maintained for 17 months through 35 passages. Morphologically, the KYN-2 cells have retained the characteristics of the original HCC, being pleomorphic and composed of various types such as cells with relatively small, polygonal, eosinophilic cytoplasm and oval-shaped nuclei with a marked tendency to pile up, flat cells with abundant clear cytoplasm and oval-shaped nuclei, and many multinucleated giant cells, proliferating in a pavement-like cell arrangement. Some junctional complexes and a number of microvilli are evident between the cells by electron microscopy. Functionally, these cells were found to secrete albumin, alpha 1-acid glycoprotein, alpha 1-antitrypsin, ceruloplasmin, transferrin, complement C, fibrinogen, fibronectin, prothrombin, retinol-binding protein (serum type), alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), ferritin and beta 2-microglobulin in chemically defined medium (CDM). The secretion of AFP and CEA is apparently dependent upon culture medium and passage. The doubling time of cells growing in serum-containing medium at the 14th passage was 84 h, and those of cells in serum-containing medium, HB101 (serum-free medium) and CDM at late passage were 28, 68, and 42 h, respectively. Chromosome analysis revealed that the chromosome number ranged from 56 to 69 without a mode, and the presence of marker chromosomes. HB virus DNA sequence was not detected by hybridization analysis. The tumorigenicity of KYN-2 cells was identified by development of tumors in nude mice after subcutaneous injection of the cells; the tumors showed an appearance basically similar to that of the original HCC. Thus, these findings suggest that the KYN-2 cell line is available as a new human HCC cell line and should be useful for various studies on HCC.

Heterotransplantation of an alpha-fetoprotein-producing human gallbladder carcinoma into nude mice.

An alpha-fetoprotein (AFP)-producing human gallbladder carcinoma showing direct invasion into the liver was transplanted into BALB/c-nu/nu nude mice. Although patient serum levels of AFP and carcinoembryonic antigen (CEA) were within normal limits, they were elevated to 1,040 ng/ml and 22.1 ng/ml, respectively, after cholecystectomy. Prominent liver metastasis was demonstrated by diagnostic imaging techniques shortly after the operation. Pathologically, the resected tumor consisted of papillotubular adenocarcinoma and the part which had invaded the liver showed a solid growth pattern with no papillo-tubular structure. The transplanted tumor showed both papillo-tubular and solid growth patterns, in which positive reactions for AFP, CEA, ferritin (FER), carbohydrate antigen 19-9 (CA 19-9), albumin (ALB) and fibrinogen (FIB) were confirmed by the avidin-biotin-peroxidase complex method. Serum levels of AFP, CEA, CA 19-9, beta 2-microglobulin (BMG) and FER were elevated in the nude mice bearing tumor transplants. Twenty-five percent of the serum AFP from nude mice with tumor transplants bound with concanavalin A (Con A), suggesting that the tumor was of gastrointestinal rather than hepatic origin.

Cornelia de Lange syndrome associated with Wilms' tumour and infantile haemangioendothelioma of the liver: report of two autopsy cases.

Two cases of Cornelia de Lange syndrome associated with infantile haemangioendothelioma of the liver and Wilms' tumour are reported. The patients showed the characteristic facies of the Cornelia de Lange syndrome, with synophrys, long curly eyelashes and small upturned nose, and physical features, including generalized hirsutism, monodactyly, syndactyly and clinodactyly. Post-mortem examination revealed annular pancreas, patency of the foramen ovale, duodenal atresia and evidence of cytomegalic infection. The cases are reported to document a possible association between malformations and neoplasms in this syndrome.

Establishment and characterization of a new human renal cell carcinoma cell line (KRC/Y).

A new renal cell carcinoma (RCC) cell line (KRC/Y) has been established from a surgical specimen of a 41-yr-old Japanese female patient with RCC composed of both clear cells and granular cells. This cell line has been maintained for more than 15 mo. through 45 passages with a stable growth. KRC/Y cells have clear or eosinophilic polygonal cytoplasm and round to oval nuclei with one or two nucleoli, and proliferate in a pavementlike cell arrangement with a lack of contact inhibition. By electron microscopy, these cells contain abundant fat droplets and glycogen granules or well-developed organelles or both, which were also observed in the original tumor. The doubling time of these cells at the 15th passage was 73 h. The chromosome number was from 37 to 45 with a hypodiploid modal number of 42. Tumorigenicity was identified by tumor formation after subcutaneous injections of KRC/Y cells in nude mice, which showed close resemblance to the original tumor by light and electron microscope observations.

Establishment and characterization of a human combined hepatocholangiocarcinoma cell line and its heterologous transplantation in nude mice.

A human cell line (KMCH-1) derived from a surgical specimen of combined hepatocellular and cholangiocarcinoma has been established. The original tumor consisted of both hepatocellular carcinoma of the trabecular type and cholangiocellular carcinoma. This cell line has been maintained for 26 months through 75 passages. KMCH-1 cells show characteristics of adenocarcinoma on light and electron microscopy. They proliferate in culture in a pave stone arrangement. The doubling time of these cells at the 24th passage was 39 hr. Chromosome analysis revealed a chromosome number ranging from 60 to 98, with a modal number of 74. KMCH-1 cells produced tumors several months after subcutaneous and intraperitoneal transplantations into athymic nude mice. Histologically, the subcutaneous tumors were poorly differentiated adrenocarcinoma, while intraperitoneal tumors were moderately to well-differentiated adenocarcinoma. Hepatocellular carcinoma components were not observed. Thus, KMCH-1 cells demonstrate the feature of cholangiocellular carcinoma in vitro and in vivo. This KMCH-1 cell line is the first established combined hepatocellular and cholangiocarcinoma cell line and may contribute to further investigation of combined hepatocellular and cholangiocarcinoma.