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Objective. High levels of interleukin-6 (IL-6) have been found in the synovial fluid of patients with rheumatoid arthritis (RA). We undertook the present study to investigate the role of IL-6 in this disease. Methods. We examined the effects of IL-6, in comparison with IL-1, on the biosynthesis of extracellular matrix macromolecules and of matrix-degrading proteinases in rheumatoid synovial fibroblasts. Results. In rheumatoid synovial fibroblasts, IL-6 by itself enhanced the production of plasminogen activator, its inhibitor, and tissue inhibitor of metalloproteinases, whereas it did not modulate the biosynthesis of precursor of matrix metalloproteinase 1 (proMMP-1) (tissue collagenase), proMMP-3 (stromelysin), or connective tissue components. However, IL-1–induced production of proMMP-1 and proMMP-3 was preferentially augmented by IL-6. Conclusion. These results suggest that in RA, IL-6 may participate along with IL-1 in fine tuning of the catabolism of connective tissue components, by modulating the balance between connective tissue–degrading enzymes and their inhibitors.

A rare case is reported of malignant mesenchymoma of a retroperitoneal lesion involving the kidney and composed of leiomyosarcoma and liposarcoma. The tumor, measuring 13 x 12 x 11 cm and weighing 1910 g, arising from the soft tissue of the retroperitoneum or renal capsule, was completely removed surgically. Thirteen months after surgical operation, multiple metastasis was observed. Immunohistochemical and electron microscopical examination showed two different mesenchymal elements. Magnetic resonance imaging findings were compatible with the pathological findings. This case appears to be the second in the world literature.

Three cases of medulloblastoma characterized by epithelial differentiation are described in patients 6-months-, 1-month- and 8-years-old. Histologically, tumors from the two infant patients showed a perivascular arrangement without apparent radiated cytoplasmic processes from the vessels. Tumor cells displayed round and/or pleomorphic vesicular nuclei and a more abundant eosinophilic cytoplasm than that found in classic medulloblastoma. Neither Homer-Wright rosettes nor ependymal or ependymoblastic rosettes were noted in these tumors. The tumor in the 8-year-old patient exhibited a classic medulloblastoma component intermingled with abundant eosinophilic cytoplasm forming a tubular structure. Immunohistochemically, tumor cells in all cases were positive for cytokeratin, synaptophysin, and vimentin. In the third case involving the 8-year-old patient, epithelial tumor cells were positive for cytokeratin, whereas classic medulloblastoma components were negative for cytokeratin. Positive staining for melanoma-specific antigen was seen only in the third case, where strong reactivity of tumor cells formed a tubulus. However, the classic medulloblastoma component was negative for melanoma-specific antigen. Ultrastructurally, basal laminae were observed around tumor cells in the 6-month-old patient. These morphological and immunohistochemical features suggest that medulloblastoma with epithelial differentiation is a rare but distinct variant of medulloblastoma, and that some of these tumors should show differentiation in ocular pigment epithelium.

In response to vascular insults, inflammatory cytokines stimulate vascular smooth muscle cells (SMCs) to express an inducible isoform of nitric oxide synthase (iNOS). Asymmetric dimethylarginine (ADMA), an endogenous NO synthase inhibitor, is metabolized by dimethylarginine dimethylaminohydrolase (DDAH). To determine whether the ADMA-DDAH system regulates cytokine-induced NO production, cultured rat SMCs were exposed to interleukin-1β (IL-1β). IL-1β (1 to 100 U/mL) dose-dependently stimulated not only iNOS but also DDAH expression and enzyme activity, accompanied by an increase in NO metabolite and by a decrease in ADMA content in culture media. A DDAH inhibitor (4124W, 5 mmol/L) augmented ADMA production ( P P

To understand the role of the cell cycle regulatory protein in the control of smooth muscle cell (SMC) proliferation, we tested the overexpression of p21Waf1, a cyclin-dependent kinase inhibitor, in human normal (MS9) and immortalized SMCs (ISS10) transfected with ori-minus simian virus 40 DNA, using an adenovirus-mediated system. In MS9, overexpression of p21Waf1 resulted in the inhibition of cell cycle progression at the G1/S boundary without apoptosis. On the other hand, in ISS10, overexpression of p21Waf1 induced marked apoptosis. In these cells, immunohistochemistry revealed that overexpressed p21Waf1 was localized in the nucleus. No differential expression pattern of either p53 or SV40T was observed in p21Waf1- and control gene (beta-galactosidase)-infected cells. Old-passaged ISS10 cells eventually showed growth arrest and a senescent-like phenotype. Immunohistochemistry revealed that p21Waf1 was localized in the cytoplasm of the early-passaged cells, but was found in the nucleus of the old-passaged cells. Our data suggested that nuclear accumulation of p21Waf1 plays a role in the cell death of immortalized SMC, which carries dysfunction of the cell cycle regulatory proteins such as p53. This culture model may be useful for studying the process of SMC proliferation, cell death, senescence, and cell cycle regulation.

Background —The Rho/Rho-associated kinase (Rho-kinase) system is implicated in various cellular functions, including migration, proliferation, and apoptosis. Because a possible role of the system is suggested in neointima formation after vascular injury, we sought to examine whether a new specific Rho-kinase inhibitor, Y27632, prevents neointima formation of the balloon-injured rat carotid artery, and if so, to investigate the effects of Y27632 on migration, proliferation, and apoptosis of smooth muscle cells (SMCs) in the injured artery. Methods and Results —Y27632 was administered intraperitoneally from 1 day before to 14 days after vascular injury. Treatment with Y27632 inhibited phenylephrine-induced Rho-kinase activation in the carotid artery on the basis of immunoblotting against the phosphorylated myosin-binding subunit of myosin phosphatase. Y27632 markedly prevented neointima formation at days 7 and 14. In controls, BrdU + proliferating and TUNEL + apoptotic SMCs were transiently and coincidentally increased in the neointima, with a peak at day 7. Y27632 significantly increased the neointimal TUNEL + SMCs at days 7 and 14, but not BrdU + SMCs. Y27642 significantly decreased the number of intimal SMCs at day 4, while not affecting the number of BrdU + or TUNEL + SMCs. Reendothelialization after balloon injury was not significantly affected by Y27632 at days 7 and 14. Conclusions —Y27632 inhibited neointima formation by enhancing SMC apoptosis and probably by suppressing early SMC migration. Therefore, a role of Rho-kinase is suggested in neointima formation after vascular injury.

A rare case of cardiac malignant fibrous histiocytoma (MFH) is reported. A 55-year-old woman complained of palpitation due to atrial fibrillation. Echocardiography, magnetic resonance imaging, and angiography demonstrated a tumor arising from the posterior wall of the left atrium. At surgery, the tumor was almost entirely resected and histologically defined as MFH. Neither chemotherapy nor irradiation was administered. Echocardiography revealed a local recurrence two months after the surgery and the patient died of advanced cachexy and heart failure 2 years later. The details of this case are presented with a review of the literature.

The expression of matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) by human vascular smooth muscle cells (SMC) was monitored as a function of the phenotypic modulation in vitro. Cell phenotype was manipulated by varying serum concentration and cell density. Synthetic phenotype was characterized by a minimum expression of the contractile proteins and a maximal proliferation rate. Contractile phenotype was quiescent and expressed a maximal level of contractile proteins. Synthetic cells expressed the highest levels of both MMP-1 and TIMP-1 and displayed maximal collagenolytic activity. No significant change was detected in MMP-2 expression or catalytic activity. Enzyme immunoassays revealed that MMP-1 expression fell by 77+/-2.4-95+/-0.5%, and that of TIMP-1 by 34+/-0.5-59+/-1.9%, as the cells acquired a contractile phenotype. The level of the MMP-1/TIMP-1 complex was similarly reduced by 78+/-2.9-85+/-1.6%. These data demonstrate that the expression of MMP-1 and TIMP-1 are coordinately regulated with SMC phenotype.

The angiomatous variant of the pleomorphic xantho-astrocytoma (PXA) is a morphological subtype of PXA that is characterized by the presence of large numbers of tiny blood vessels with variable wall thicknesses. A case of an angiomatous variant of PXA occurring in the temporal lobe of a 43-year-old man is presented. The patient had a 20-year history of seizures. Considering the clinical course and intra-operative electro-encephalography findings, it appears that this tumor was the epileptic focus in this patient and that it had been present for 20 years. Histopathologically, various degenerative changes, such as microcystic changes, calcification and hyaline thickening of vessel walls were seen in the tumor. In particular, obvious vascularity in the tumor and secondary vascular degenera-tion should be related to gradual growth and benign clinical course of an angiomatous variant of PXA.

Cyclin-dependent kinase inhibitor p21Waf-1 is recognized as a negative regulator of cell cycle progression, and it possibly mediates cell differentiation and apoptosis. To understand the role of p21Waf-1 in phenotypic modulation of vascular smooth muscle cells (SMC), we induced the overexpression of p21Waf-1 in cultured rat SMC. The recombinant adenovirus vector encoding p21Waf-1 (AdvCMVp21) was constructed by homologous recombination and the vector encoding beta-galactosidase (AdvCMVLacZ) was used as an experimental control. Administration of AdvCMVp21 suppressed serum-induced proliferation and cell cycle progression; however, the number of quiescent cells and the population of TUNEL-positive cells were not altered. Overexpression of p21Waf-1 did not affect the expression of contractile proteins and the availability of an endogenous growth factor signal p21Waf-1 may regulate cell cycle progression in SMC without affecting the apoptotic process and cell differentiation. Furthermore, the longitudinal diameter of AdvCMVp21 infected cells was increased compared with that of AdvCMVLacZ infected cells. Total protein content was also increased in AdvCMVp21 infected cells. Responses to the serum stimulation, proliferation and total protein synthesis may be independently regulated. Thus, the suppression of cell cycle progression by p21Waf-1 resulted in cellular hypertrophy of SMC.

We report a case of dedifferentiated leiomyosarcoma. The patient was a 56 year-old man. The tumor was local recurrence in the right greater pectoral muscle after radiotherapy and chemotherapy. Tumor showed typical moderately leiomyosarcoma with dedifferentiated component. This component was revealed as pleomorphic malignant fibrous histiocytoma as a result of immunohistochemistry. These results indicated dedifferentiated components derive from loose muscle features due to dedifferentiation. We diagnosed this case as dedifferentiated leiomyosarcoma.

We describe a rare case of pleomorphic type of malignant fibrous histiocytoma (MFH) in the buttock that presented a systemic involvement. The case was of a 58-year-old woman presenting hepatic dysfunction and inflammatory reactions including fever, positive C-reactive protein (CRP), an elevated erythrocyte sedimentation rate, and high levels of platelets and ferritin. The fever of 3 months duration subsided on the first postoperative day. The MFH resection also brought rapid normalization in CRP, platelets, and leukocytes. The local and systemic productions of cytokines induced by this tumor were evaluated. In vivo and in vitro production of interleukin (IL)-6, IL-1beta, and tumor necrosis factor alpha by tumor cells were measured using enzyme-linked immunosorbent assay. Blood samples taken preoperatively, tumor tissues, and the primary culture medium showed extraordinarily high IL-6 levels. The plasma IL-6 level was normalized postoperatively. Immunohistochemistry showed the positivity of tumor cells for IL-6. The IL-6 produced by the tumor was concluded to have been responsible for the systemic illness.

口腔粘膜の扁平上皮癌, および上皮の増殖性病変の細胞診において, 歯間ブラシによる細胞採取法が有用であるか, 綿棒採取法との比較検討を行った.対象は扁平上皮癌 (高分化型65例, 中等度型7例, 低分化型1例), 良性病変および自然治癒47例で行った.良悪性問の細胞数は歯間ブラシによる採取では綿棒の約2.5倍の細胞数の増加がみられ, 細胞集塊数では約4.7倍の集塊数がみられた.腫瘍径による細胞集塊出現数ではT2 (2.1～4cm) はT1 (2cm以下) の約1.2倍の細胞の増加がみられ腫瘍径が大きくなるほど細胞数が多くなった.肉眼的腫瘍発育形態による細胞数については潰瘍型, 硬結型, 乳頭型, 白斑型, 肉芽型, いずれも2～3倍の細胞数の増加がみられ, 特に外向性発育を示す白斑型, 肉芽型において顕著であった.歯間ブラシによる採取法の利点としては第一に細胞数の増加, 第二に細胞集塊として採取されるため, 腫瘍細胞間の極性の乱れ, 核間距離の不整, 細胞密度が観察可能な点があげられる.さらに細胞分布は綿棒採取法に比べ均一に塗抹される場合が多く, 壊死物質や炎症細胞に重なった細胞の見落としを防ぐことができた.以上の結果より, 口腔病変, 特に上皮の増殖性病変における細胞診断に歯間ブラシは有用と考えられた.

The purpose of this study was to investigate the mechanism of cell death in chondrocytes of the growth plate. In the degenerative chondrocyte zone of the growth plate, apoptotic chondrocytes were defeated by the in situ nick end labelling method, by DNA analysis in agarose gel, and by electron microscopy. The results of the in situ nick end labelling method and the occurrence of a ladder pattern of DNA in agarose gel analysis indicated the activation of endogenous endonucleases, resulting in DNA fragmentation. Electron micrographs showed the early morphological changes associated with apoptosis. This report presents both morphological and biochemical evidence for apoptosis in the terminal hypertrophic chondrocytes of the growth plate. These data suggest that apoptosis of degenerative chondrocytes may play an important role in the control of normal and pathological endochondral ossification.

Nitric oxide (NO) inhibits platelet adhesion and aggregation in vitro. In vivo, chronic inhibition of NO synthesis induces nephrosclerosis and hypertension. Although the pathophysiological mechanism of this glomerular injury has not been clarified, sympathetic nerve activation, a potent procoagulant stimulus elicited by NO inhibition, may play a role. To investigate the role of renal sympathetic nerves in the development of renal injury induced by NG-nitro-L-arginine methyl ester (L-NAME), a specific NO synthesis inhibitor, we examined renal histological changes in four groups of Sprague-Dawley rats: (1) sham operated, vehicle treated; (2) sham operated, L-NAME treated; (3) denervated, vehicle treated, and (4) denervated, L-NAME treated. Following renal denervation or sham operation, L-NAME was administered orally for 4 weeks. Chronic NO inhibition induced platelet aggregation and erythrocyte stasis in the glomerular capillary lumen accompanied by electron-microscopic glomerular injury. Renal denervation abrogated platelet aggregation and glomerular injury in L-NAME-treated animals. Thus, chronic NO synthesis inhibition induced intraglomerular platelet aggregation and glomerular injury, which was attenuated by renal nerve denervation. These results suggest that intrinsic NO may have an antithrombotic effect in the glomeruli and may play a protective role in the progression of glomerular injury possibly mediated by renal sympathetic nerves.

乳腺の細胞診延べ2,741例を病理組織診と対比し, 細胞診にてClass III以上と診断し乳癌が疑われた症例で, 画像診断上悪性所見を示さず細胞診のみ乳癌の術前診断を行い得た症例の病理形態学的特徴について検討をした。細胞診のクラス分類の内訳としてClass Iは1,470例 (53.6%), Class IIは131例 (4.8%), Class IIIは59例 (2.2%), Class IVは41例 (1.5%), Class Vは224例 (8.2%), さらに検体不良例816例 (29.8%) であった。細胞診にてClass III以上と診断した324症例のうち病理組織診断が施行され, 術後の再発例を除き悪性と診断されたのは234症例であった。234症例の乳癌のうち画像診断上悪性所見の認められなかった症例は17症例 (7.3%) であった。画像診断上悪性所見の認められなかった症例の組織型は, 乳頭腺管癌8例, 粘液癌4例, 硬癌2例, 充実腺管癌1例, 嚢胞内乳頭癌1例, 非浸潤性乳管癌1例であり, 腫瘤の大きさとしては1.0cm以下が9例, 1.1～1.5cmが8例と1例は乳頭分泌物で腫瘤は認められず, 全症例とも1.5cm以下の腫瘤であった。さらに粘液癌では1.5cm以下の大きさの腫瘤ではすべて画像診断上は悪性所見に乏しく, 細胞診のみが乳癌の術前診断に有用であった。

The gene expression of two type IV collagenases (matrix metalloproteinase [MMP]-2, a 72 kd type IV collagenase, and MMP-9, a 92 kd type IV collagenase) was investigated in carcinomas of the hypopharynx. We examined 27 cases operated on in our hospital by an in situ hybridization technique to detect their messenger RNA signals in cancer cells and surrounding stroma. Both signals were detected in all cancer nests and in stromal cells in the same specimens. Clinicopathologic studies showed a significant relationship between MMP-2 expression in the primary cancer and the outcome of treatment. Our present study suggests that hypopharyngeal squamous cell carcinoma producing MMP-2 has a high potential for invasion and metastasis and a poor outcome. The analysis of MMPs will be useful for treatment planning in hypopharyngeal carcinoma and for prognosis.

Granular cell tumors (GCTs) are rare, soft tissue tumors, and their histogenesis is still controversial, although the neurogenic theory, especially Schwann cell derivation, is considered the most plausible at present.We studied eight cases of GCT clinicopathologically and immunohistochemically. The results were as follows:1. Granular cells of GCT had a variety of sizes and shapes, were filled with fine eosinophilic granules, and showed lysosome-like characteristics, also with a variety of sizes and shapes, on electron microscopic observation.2. Immunohistochemically, the intracytoplasmic granules showed positive reactions for S-100 protein, NSE, Kp-1 and diastase-resistant PAS staining but a negative reaction for a 1-antichymotrypsin. Additional lipid staining, such as oil red O, Sudan black B, and Nile blue, was performed in two cases, but was all negative.3. In three cases, an appearance of transition from striated muscle fibers to granular cells, and the accumulation of intracytoplasmic granules with the shape of striated muscle fibers remaining were seen at the margin of GCT masses. In addition, four cases out of eight showed weakly positive reactions for myoglobin.Thus, several cases were suggested to have a myogenic origin, and neither 5-100 protein nor NSE provided conclusive evidence of neurogenic origin due to their poor specificity. It was, therefore difficult to limit their derivation to neurogenic origin, and further examinations are required.The accumulation of intracytoplasmic granules in other tumors, such as neurofibroma, which was referred to as granular cell neurofibroma by Enzinger and Weiss (1983), and leiomyoma, were previously reported. The neoplastic proliferation of granular cells seems unlikely in view of their few cell organelles on electron microscopic observation.In order to explain our results and those of previous studies, it is most reasonable to consider that GCT is not a neoplastic lesion, that the accumulation of intracytoplasmic granules in GCT is the result of degenerative or reprocessing changes (Whitten, 1968), and that such “granular cell changes” occur in various tissues (Shear, 1960 and Millar, 1977).Furthermore, previously reported “malignant” GCTs, which showed histological malignancy and remote metastasis and were often diagnosed with much difficulty, may include pre-existing malignant tumors with “granular cell changes”.

Five patients with malignant bone tumors, [four osteosarcomas and one malignant fibrous histiocytoma (MFH)] were treated by adjuvant chemotherapy combined with caffeine. In two patients who were treated pro-operatively, the tumors showed histologically 100% or 80% necrosis. In one of two patients with lung metastasis, the chemotherapy resistant tumor did show a reduction in size on computed tomography (CT). All patients reported side effects, especially hyperventilation syndrome in two patients, which may have been a direct side effect from the caffeine. The results suggested that chemotherapy combined with caffeine was more effective for treating malignant bone tumors, although more care is needed to control some side effects associated with this regimen.

Although the Angio-Seal arterial closure device is widely used for preventing bleeding and facilitating early ambulation after arterial puncture, it is also associated with unique complications, such as stenosis, occlusion, or peripheral embolism. We report the first case of a foot ulcer that developed 70 days after an Angio-Seal application. The collagen sponge component accidently positioned itself in the arterial lumen and was not absorbed. A foreign body reaction was observed microscopically. In patients with arteriosclerosis, the Angio-Seal device should be used carefully; post procedural monitoring is necessary after implantation.

We investigated the relationship between pathological findings and MR images of 5 resected soft tissue tumors. These included desmoid, hemangioma, liposarcoma, schwannoma and malignant schwannoma. The increase of collagen was an important histological feature of tumors, and this was seen on MRI as a decrease in signal intensity, especially, on T2 weighted images. Minute amounts of fat tissue appeared as high intensity on T1 weighted images on MRI. This study suggested that MRI may reflect minute histological changes in the matrix of soft tissue tumors.

Two patients with neuroblastoma of the central nervous system are described. The first, a 10-month-old male infant with symptoms of vomiting and weakness of the right upper extremity, was found on magnetic resonance imaging (MRI) to have enhanced mass lesion in the left hemisphere, vermis of the cerebellum and part of brain stem. The second, a 3-year-old boy who had swelling of the right side of the head, was found on computed tomography (CT) to have enhanced mass lesion of the frontal hemisphere and frontal bone. Histological examination of the surgical specimen of both cases demonstrated neuronal differentiation. The clinicopathological features of cerebellar neuroblastoma in comparison with metastatic neuroblastoma of the frontal lobe were discussed.

We present our data on the cytotoxicity of microtubule inhibitors (paclitaxel and taxotere) and their mode of action against cisplatin-sensitive or -resistant ovarian cancer cell lines in vitro. To try to establish an optimal administration schedule in clinical use, the cytotoxicity of microtubule inhibitors alone and in sequential combination with cisplatin was investigated. With microtubules alone, a marked dose- and schedule-dependent growth inhibition was demonstrated in lower concentrations and only schedule-dependency in higher concentrations. The cytotoxicity of taxotere was approximately 0.8- to 132-fold that of paclitaxel. No cross resistance to cisplatin was observed. In the relation between assay AUC (area under the concentration curve) and growth inhibition, increasing AUC by dose escalation seemed not to be advantageous. In a combination with cisplatin, the treatment of microtubules over 48 h followed by cisplatin administration demonstrated the most effective cytotoxicity in cisplatin-resistant cell line.

We conducted a 20-year retrospective survey of 6.457 autopsied cases to select those women who died of primary ovarian carcinoma. Our objective was to determine the incidence and the sites of the metastases in those treated with or without combination chemotherapy based on cis-diamine dichloride platinum (CDDP). In the 70 cases so identified, epithelial ovarian tumors were the most common, found in 52 of the 70 cases. There were 14 cases of germ cell tumors and 2 cases each of sarcoma and stromal cell tumors. Metastases involved the peritoneum in the majority of cases (87%). The liver was the next most frequently affected (73%), followed by the spleen (47%) and the lung (44%). Of the 70 cases, 33 (47%) had received CDDP, while a regimen of 5-fluorouracil, doxorubicin, mitomycin C and toyomycin (FAMT) was administered to 11 cases (15.7%) and radiotherapy to 10 cases (14.3%). Patients who received a CDDP regimen survived longer (23.4 months) than those treated with FAMT (19.2 months) or with other regimens (13.6 months). The most common cause of death was carcinomatosis (48/70 cases). The high incidence of pulmonary, hepatic and splenic metastases in women with primary ovarian cancer indicates the need to direct anticancer treatment toward those sites to thus improve survival.

The secretion of matrix metalloproteinases (MMPs), which enzymes have been suggested as degradative agents for components of the extracellular matrix, was studied in eight glioma cell lines. We investigated the relationship between the secretion of MMPs and the tumorigenicity of the glioma cells by immunoblot analysis using antisera against human MMPs. Four types of matrix metalloproteinase including MMP-1, MMP-2, MMP-3 and MMP-9 were detected in the glioma cells. MMP-2 was detected in all high-grade gliomas, while an additional MMP or two were detected in only two cell lines, U87MG and KINGS-1. Furthermore, the tumorigenetic potential was shown to correlate with the secretion of MMPs in vitro. These enzymes are capable of degrading several extracellular matrix components of the central nervous system, and thus must play an important role in the mechanism of occurence and/or growth of glioma.

According to the depth of invasion, sixty-one superficial esophageal cancers (SECs) were subclassified as follows: intraepithelial cancer (ep), mucosal cancer invading to the lamina propria mucosa (lpm), or to the muscularis mucosa (mm), and submucosal cancer limited to the inner one-third of the submucosal layer (sm1), to the middle one-third (sm2), or invading into the outer one-third (sm3) to investigate the relationship between the depth of invasion and gross type classified according to the Guidelines of the Japanese Research Society for Esophageal Disease. Our results show that the SECs comprised of 9 ep cancers, 8 lpm, 6 mm, 6 sm1, 19 sm2 and 13 sm3, none of the ep or lpm cancers had either lymph node metastasis or recurrence and one-third of the mm and sml cancers had metastasis only in the mediastinal nodes. One-third of the sm2 and sm3 cancers, otherwise, revealed nodal involvement at surgery, and one-fourth of those recurred postoperatively. Recurrence in the cervical nodes was found in 9% of these cancers. All the SECs of the gross types including protruding type, slightly elevated type more than 2 cm in diameter and distinctly depressed type were submucosal cancers, whereas all the SECs showing the gross types including flat type and slightly elevated type less than 2 cm in diameter were ep or lpm cancers. Therefore, we concluded that for SEC evaluated as flat or slightly elevated type less than 2 cm in diameter by the endoscopic examination, less radical treatment such as endoscopic mucosal resection (EMR) should be performed at first.

The expression of matrix metalloproteinase-2 (MMP-2, type IV collagenase, 72 kDa) in 47 cervical precancerous and cancerous lesions were localised by in situ hybridization and immunohistochemical studies. MMP-2 mRNA was present in the stromal cells surrounding invasive cells in the majority of squamous cell carcinomas and adenocarcinomas. In contrast, MMP-2 immunoreactivity was found in the epithelial cells. Our results indicate that there may be some interaction between carcinoma cells and stromal cells to produce MMP-2 which may contribute to the destruction of the extracellular matrix in invasive cervical carcinomas.

We investigated the effects of epidermal growth factor (EGF) and transforming growth factor-beta (TGF-beta) on cell growth and on regulation of matrix metalloproteinases (MMPs) in four cell lines of human esophageal squamous cell carcinomas (TE8, TE9, TE10, and TE11). EGF stimulated the production of proforms of gelatinase B (MMP-9) by three cell lines that could synthesize EGF by themselves, with TE9 being the exception. Particularly, both the production of MMP-9 and DNA synthesis in TE10 were stimulated significantly by EGF. TGF-beta slightly stimulated DNA synthesis in two cell lines, TE9 and TE11, and TGF-beta secretion by TE9 was detected. The production of proforms of gelatinases A (MMP-2) and MMP-9 was gradually induced by TGF-beta in a concentration-dependent manner in all the cell lines except for TE9. Flow cytometric analysis revealed that all the lines expressed both EGF- and TGF-beta-receptors. In conclusion, our present results indicate that at least there are possibly two distinct phenotypes in human esophageal squamous cell carcinoma: one (TE10) depends on autocrine EGF production that enhances DNA synthesis and MMP-9 production; and the other (TE9) on autocrine TGF-beta that stimulates DNA synthesis but not in relation to gelatinase production.

A cell line derived from medial smooth muscle cells (SMC) was established from the porcine coronary artery by transfection with ori-defective simian virus 40 plasmid DNA (SV40 DNA). The characteristics of transfected cells (SV40-SMC) such as cell growth, collagen and non-collagen syntheses were investigated. SV40-SMC expressed SV40 large T antigen, c-myc and c-myb encoded proteins in the nuclei. SV40-SMC demonstrated a 'hills and valleys'-like arrangement in overconfluence and actin filaments upon immunofluorescent staining. Under electron microscopic observation, SV40-SMC had larger amounts of synthetic organelles and smaller amounts of filament bundles than those of SMC. SV40-SMC demonstrated three times higher growth activity and 4.4 times greater cellular density than SMC. Smooth muscle cells did not grow in media containing 5% plasma derived serum (PDS) instead of normal serum, whereas SV40-SMC proliferated in this medium. SV40-SMC did not grow in soft agar gel, while HeLa S3 cells, a cell line of human cervical carcinoma, formed colonies in this gel. By immunofluorescent (IF) staining, collagen phenotypes I, III, IV and V were detected in both SV40-SMC and SMC. However, protein synthesis including collagen and non-collagen was higher in SV40-SMC than in the control sample. It was concluded that SV40-SMC were a continuous cell line for vascular SMC regarding morphological characteristics, and demonstrated a higher growth activity, with increased collagen and non-collagen syntheses. This cell line is useful for the investigation of atherogenesis in relation to a proliferation of SMC and an accumulation of extracellular matrices in vascular intima.

To investigate whether lipid peroxides in tissue and/or blood play an important role in the injury to mesangial cells and glomerular structural proteins, we studied the effect of linoleic acid hydroperoxide (LHO) on mesangial cells cultured from rat kidneys. When 1.0-3.0 nmol/ml of LHO were added to the culture, the production of 70-kDa gelatinolytic neutral proteinase by mesangial cells was enhanced without morphological damage. The production of the enzyme, however, was inhibited by an addition of 4.0 or 5.0 nmol/ml of LHO and a microscopic examination also demonstrated marked injury to mesangial cells in the presence of these concentrations of LHO. These data suggest that there is a possibility that increased lipid peroxides in tissue and/or blood play a role in the injury to mesangial cells and glomerular structural proteins leading to mesangiopathy.

To investigate the effect of matrix-degrading enzymes on the malignant potential of oral squamous cell carcinoma, the expression of matrix metalloproteinase-2 (MMP-2/72-kD gelatinase/type IV collagenase) in 46 patients who had neck surgery for oral cancer was studied immunohistochemically. In 20 of 26 patients (76.9%) with lymph node metastases, proMMP-2 was strongly expressed, whereas the production of proMMP-2 in tissue was detected only in 5 of 20 patients (25%) who had no lymph node metastases. In tissue specimens, proMMP-2 was expressed in a diffuse invasive mode and in the advancing front of cancer. Because MMP-2 can degrade type IV collagen composed of basement membrane, these results suggest that the in vivo production of the enzyme by cancer is an indicator of the degree of malignancy, and that the analysis of proMMP-2 expression is useful to evaluate the malignant potential in individual oral squamous cell carcinoma.

乳腺の穿刺吸引細胞診にて乳管癌が疑われた乳房部原発基底細胞上皮腫の1例の細胞像について報告した.症例は65歳, 女性, 右乳房C領域に1.5×1.5cmの腫瘤が認められ10年来大きさは不変であった.Mammography, 超音波などの画像診断にて乳管癌が疑われ外来受診時に穿刺吸引細胞診が施行された.細胞診所見は出血像を背景に腫瘍細胞は大小の結合性の強い細胞集塊で出現し, 細胞の配列は不均等で細胞の重積性が認められた.核は類円形小型で核クロマチンの増量がみられ, 乳管癌も否定できず術中迅速病理組織診断が行われた.病理組織にて基底細胞上皮腫と診断された.基底細胞上皮腫の一般的な細胞学的所見としては大小の細胞集塊での出現, 細胞は小型類円形で大小不同性に乏しく, 細胞集塊の辺縁に柵状配列が認められることであると思われるが, 乳房部に発生する基底細胞上皮腫は非常にまれであり, 本症例では術前の穿刺吸引細胞診での確定診断は困難であった.

Background. Matrix metalloproteinases (MMP) are a gene family of zinc enzymes capable of degrading almost all of the extracellular matrix macromolecules in vivo. Their enzymic activities are believed to be responsible for tumor invasion and metastasis. Methods. In this study, using peroxidase-antiperoxidase method, monospecific antisera against MMP-1 (tissue collagenase), MMP-2 (type IV collagenase/72-kilo-dalton [KD] gelatinase), and MMP-3 (stromelysin) were applied to 29 squamous cell carcinomas and normal epithelium of the esophagus to identify cells synthesizing and secreting these enzymes. Results. Immunoreactivity of MMP-1, −2, and −3 was observed in small cancer nests of the deeply invasive or marginal portion of the tumor. Among the 29 patients studied, the presence of at least one MMP was observed in 17 (58.6%). All three enzymes were observed in six (20.6%) patients, MMP-2 and −3 in five (17.2%) patients, only MMP-2 in three (10.3%) patients, and MMP-3 alone in three (10.3%) patients. There was a good correlation among histologic stage and tumor invasion, lymph node metastasis, and MMP expression. In particular, expression of MMP-2 and −3 was closely related to lymph node metastasis and vascular invasion. Conclusions. These results suggest that MMP, especially MMP-2 and −3, play an important role in tumor invasion and metastasis and that analysis of MMP-2 and −3 production is useful for evaluation of malignant potential in esophageal carcinoma. Cancer 1992; 702747–53.

A case of a dentigerous cyst associated with a deciduous tooth in a 2-year-old boy is presented. Radiologic examination revealed a well-defined radiolucency in the right maxilla associated with the crown of a maxillary second deciduous molar. The cyst cavity was lined with nonkeratinized squamous epithelium, and odontogenic epithelial islands were seen in subepithelial connective tissue.

To investigate whether matrix metalloproteinases (MMPs) play an important role on the malignant potential of oral carcinoma, the expression of proMMP-2 and 3 in 47 patients with oral squamous cell carcinoma was analized by immunohistochemical staining of tissue specimens using monospecific antisera against human proMMP-2 and MMP-3. In 21 (77.8%) of 27 patients with lymph node involvement, the increased-production of pro-MMP-2 and 3 was observed, and proMMP-2 or 3 alone was observed in 4 patients (14.8%). On the other hand, the production of proMMP-2 and 3 in tissue was detected only in 4 (20%), and proMMP-2 or 3 alone in 7 (35%) of 20 patients, who had no lymph node metastasis. In the tissue specimens, proMMP-2 and 3 was expressed particularly in the diffuse invasive mode and in the part of advancing front of cancer. Since MMP-2 and 3 can degrade the extracellular matrix macromolecules including basement membrane components, these results indicate that analysis of proMMP-2 and 3 expression in tissue is useful for evaluation of the metastatic potential in oral squamous cell carcinoma.

食道粘膜下層癌 (以下sm癌) の悪性度を, sm層への浸潤度, 発育浸潤形式, matrix metalloproteinase-2 (MMP-2) の発現より検討した.対象症例は31例で, 1) 浸潤度 (sm1: 6例, sm2: 15例, sm3: 10例) では, sm3の80%が0に1型, sm1では癌肉腫を除き全例0-IIa型であった.また, sm3にn (+), ly・v, 再発が多かった.2) 発育形式 (下方発育型: 13例, 表層発育型: 8例, 混合型: 10例) では, 下方発育型に0-I型が, 表層発育型に0-IIa型が多かった.n (+) は下方発育型と混合型に, v (+) と再発は表層発育型に多かった.浸潤形式 (G1: 4例, G2: 12例, G3: 10例, G4C: 4例, G4D: 1例) では, G3以上に0-I型が多く, n (+) 頻度も高かった.3) MMP-2発現を9例 (29%) に認め, ly (+) と再発が多かった.以上より, 1) 0-I型は高度sm浸潤, 下方発育型, G3以上が多く, n (+), ly・vも高頻度で, 進行癌に準じた郭清が必要である.2) sm2以下でも表層発育型やMMP-2発現例は再発が多く, 強力な合併療法と厳重な経過観察が必要である.

Mortality of cancer is the result of regional and/or systemic metastasis. Matrix metalloproteinases (MMPs) were believed to play an important role in its malignant behavior. MMPs have been identified as matrix-degrading enzymes, such as procollagenase (proMMP-1) of Mr=53, 000, progelatinase (proMMP-2) of Mr=72, 000 and proMMP-9 of Mr=92, 000, and prostromelysin (proMMP-3) of Mr=59, 000. The regulatory mechanism of these enzymes is still unclear; however, it has been indicated that many growth factors such as EGF, TGF-α, TGF-β, PDGF, and etc. will play an important role in MMPs synthesis of cancer cells.In this paper, we report that two oral cell lines of squamous cell carcinoma (SCC9 and SCC25) produce at least two types of MMPs, which have been identified as proMMP-2 and 3 by the indirect immunofluorescence technique, immunoblot analysis, and gelatin-substrate gel zymography. Additionally, a 92-kDa gelatinolytic metalloproteinase was detected in SCC25 by gelatin-substrate gel zymography. Furthermore, after the treatment with EGF, the production of proMMP-1 and 3 by SCC25 was increased in a dose-dependent manner, but not by SCC9. However, unlike EGF, TGF-β stimulated both cell lines to produce proMMP-2. These results suggest that oral carcinoma cells have a potential to degrade extracellular matrix macromolecules including basement membrane components, and EGF and TGF-β are important factors for influencing the metastatic potential of oral squamous cell carcinomas.

Human aortic endothelial cells, isolated at autopsy from a 52-year-old male dying from lung cancer, were treated with simian virus 40 (SV40). One colony was isolated from the infected endothelial cell culture 4 weeks after infection. The cells expressed SV40 large T antigen and p53 protein (p53) in their nuclei but lacted the characteristics of a transformed phenotype. The cells grew well in a monolayer over the 97th passage and exhibited Factor VIII-related antigen, Ulex europaeus 1 agglutinin (UEA-1) as endothelial cell markers, and a well-developed fibronectin network. The amount of prostacyclin synthesized by the cells was less than the amount synthesized by normal aortic or umbilical cord vein endothelial cells. The cells produced relatively large amounts of procollagenase, and 12-o-tetradecanoyl-phorbol-13-acetate (TPA) augmented the ability of the cells to produce this enzyme. These immortalized human aortic endothelial cells, which have some characteristics of normal endothelial cells and, like capillary endothelial cells, have the ability to produce collagenase, will probably prove useful for studies of atherosclerosis and angiogenesis.

We performed a long-term prospective study on venous invasion of colorectal cancer. The degree of venous invasion was divided into four stages (V0 through V3). Venous invasion was classified into three types by location (Vx, Vy, and Vz). Hepatic metastasis occurred in 27%, 33%, and 20% of patients with V2, V3, and Vz tumors, respectively. Local recurrence occurred in 33% and 15% of the V3 and Vz groups, respectively. However, there were no significant differences among the groups in terms of the rate of pulmonary metastasis. The 6-year survival rate for Dukes' stage B tumors was 94%, 88%, and 74% in the V0, V1, and V2 groups, respectively. There was a significant difference in the survival rate between patients with V0 tumors and V2 and V3 tumors. However, no significant difference was noted in the location. In Dukes' stage C tumors, on the other hand, the survival rate was 77%, 56%, and 44% in the V1, V2, and V3 groups, respectively. Also, it was 85%, 73%, and 45% in Vx, Vy, and Vz cases, respectively. Significant differences were noted between V1 and V3 (or V2), and between Vz and Vx (or Vy). It appears that the degree and location of venous invasion influence not only hepatic metastasis, local recurrence, and survival rates but also have prognostic value.

The effect of linoleic acid hydroperoxide on replication of adenovirus type 2 DNA in cultured endothelial cells of bovine aorta was studied to elucidate the deleterious effect of lipid peroxides on replication of DNA in the arterial endothelial cells. When, just after the infection, the cells were incubated with 1 nmol (measured by the thiobarbituric acid reaction and expressed in terms of malondialdehyde of the hydroperoxide, the replication of adenovirus type 2 DNA was markedly reduced. The level of the messenger RNA for adenovirus type 2 DNA polymerase was also reduced. In accordance with the reduction of the messenger RNA, the synthesis of the polymerase in the cells was reduced. The inhibition of DNA replication in the arterial endothelial cells by lipid peroxides was discussed in relation to atherogenesis.

A DNA fragment encoding the polyhedrin promoter of Autographa californica multiple nuclear polyhedrosis virus (AcMNPV strain) was constructed using overlapping oligodeoxyribonucleotides (oligos), which included the 5'-untranslated leader sequence of the polyhedrin-encoding gene. This DNA fragment was cloned into an intermediate transfer vector (pKX105) providing a unique BamHI site for the insertion of foreign genes. The Escherichia coli lacZ gene was first cloned at the BamHI site of pKX105 and the XhoI-KpnI fragment containing the lacZ gene was transferred to another plasmid vector (pEI) consisting of flanking AcMNPV sequences (pEI-lacZ). The E. coli beta-galactosidase that was produced in the infected insect cells using the recombinant virus constituted about 10% of the total cytoplasmic proteins. The pKX105 plasmid was also modified to give rise to pTT-lacZ which consisted of the lacZ gene under the control of the Rous sarcoma virus long terminal repeat promoter to facilitate rapid screening of the baculoviral recombinants in which the gene of interest was cloned under the control of the polyhedrin promoter. The efficiency of these transfer vectors was verified by obtaining high levels of expression of the adenovirus(Ad)-encoded preterminal protein (pTP) which is involved as a protein primer in the initiation of Ad DNA replication. The baculovirus-produced pTP was immunoprecipitable using rabbit polyclonal antibodies raised against a hydrophilic domain of pTP. The pTP protein was localized in the nucleus of the infected insect cells, and was biologically active in the in vitro Ad type 2 (Ad2) replication initiation assay.

New cell lines, termed as KMR-1 and KMR-2, were established fron metastatic bone tumor of human renal cell carcinoma (clear cell type). They have been passaged in vitro for about 8 months. Doubling times of KMR-1 and KMR-2 were respectively 32 and 51 hours. These cells have retained glycogen (stained by periodic acid Shiff reaction and destained by diastase) and fat drops (stained by oil red O stain procedure), which are characteristic to original tumor cells.Matrix metalloproteinase (MMP) -2 and MMP-3 were detected in the conditioned medium using SDS-PAGE immunoblotting technique. Two bands of MMP-2 (Molecular weight [Mr] of 72KD and about of 30KD) were detected from KMR-2 and MMP-3 (Mr of 57KD) was detected from KMR-1. These MMPs seem to relate the metastatic potentiality of renal cell carcinoma cells.

Incorporation of [3H]proline into collagenous proteins by cultured smooth muscle cells from rabbit aorta was found to be increased in the presence of linoleic acid hydroperoxide at its concentrations of 6-60nmol/ml, indicating that the net synthesis of collagen by these cells is increased by these concentrations of lipid peroxides. Although linoleic acid slightly increased the collagen synthesis, the effect seems to be mostly ascribable to its action to increase the proliferation of the cells.

尿中に腫瘍細胞が出現した男子尿道原発悪性黒色腫について報告した.症例は64歳男性で尿道球部に発生した悪性黒色腫であった.尿細胞診では腫瘍細胞は孤立散在性に出現し, 一部疎な結合を示す集塊で出現した細胞では移行上皮癌細胞との鑑別が困難であったが, 少数の細胞には明らかなメラニン顆粒が認められた.これら尿中の腫瘍細胞は免疫組織化学染色においてS-100蛋白が陽性を示し, 摘出腫瘤の病理組織標本ではMasson-Fontana染色陽性, 鉄染色陰性, 免疫組織化学染色ではS-100蛋白陽性さらにNSE (Neuron-specifi cenorase) も陽性であった.また, 電顕にて腫瘍細胞の細胞質に多数のmelanosomeが確認された.

Free-living amebas represented by Naegleria fowleri, Acanthamoeba and Balamutia have been known to cause fatal meningoencephalitis since Fowler and Carter (1965) reported the first four human cases. An autopsy case of a 25-year-old female with primary amebic meningoencephalitis (PAM) due to Naegleria fowleri is described. Headache, lethargy and coma developed in this patient, and her condition progressed to death 8 days after the onset of clinical symptoms. Cerebral spinal fluid examination confirmed clusters of amebas, which were grown in culture and identified as Naegleria fowleri. At autopsy, lesions were seen in the central nervous system (CNS) and the ethmoid sinus. The CNS had severe, suppurative meningoencephalitis with amebic trophozoites mingled with macrophages. This case is the first report of PAM due to Naegleria fowleri in Japan.