Histopathological predictors of regional lymph node metastasis at the invasive front in early colorectal cancer.

AIMS: In early colorectal cancer (ECC), prediction of lymph node (LN) metastasis is vital for the decision of additional surgical treatment after endoscopic mucosal/submucosal resection. The aim of this study was to determine the relationship between LN metastasis and comprehensive histopathological findings including the cancer microenvironment in ECC. METHODS AND RESULTS: Using 111 ECC cases, including 36 cases with LN metastasis, histopathological observations and immunohistochemistry for lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1), von Willebrand factor, matrix metalloproteinase-7 (MMP-7), CXC chemokine ligand-12 (CXCL12) and angiopoietin-like-4 (ANGPTL4) were conducted. Relationships between LN metastasis and growth pattern, status of muscularis mucosae, depth of cancer invasion, overall histopathological type, histopathological type at the invasive front, tumour budding, neutrophil infiltration in cancer cells (NIC), fibrotic cancer-stroma type, Crohn's-like lymphoid reaction, microscopic abscess formation and lymphatic invasion were determined. In addition, the expression of MMP-7, CXCL12 and ANGPTL4 in cancer cells at the invasive front were also considered in the context of LN metastasis. By multivariate analysis, lymphatic invasion, NIC and MMP-7 expression at the invasive front were independent predictors of LN metastasis. CONCLUSIONS: LN metastasis is regulated not only by the characteristics of cancer cells but also by microenvironmental factors of lymphatics and neutrophils, especially at the invasive front.

Basic fibroblast growth factor induces down-regulation of alpha-smooth muscle actin and reduction of myofibroblast areas in open skin wounds.

To examine the effects of basic fibroblast growth factor (bFGF) on the inhibition of alpha-smooth muscle actin (alpha-SMA) expression in dermal fibroblasts, we have established two dermal myofibroblastic cell lines positive for alpha-SMA (rat myofibroblasts [RMF] and rat myofibroblast-like [RMFL] cells) and one fibroblastic cell line negative for alpha-SMA (rat fibroblasts cells) as a model of fibroblast differentiation. In contrast to the increased expression of alpha-SMA in RMF and RMFL cells, irrespective of transforming growth factor-beta1 treatment, bFGF induced a decrease in alpha-SMA expression in the myofibroblastic cells and the reduced expression patterns of alpha-SMA differed between cells, as demonstrated by Western blot and reverse transcription polymerase chain reaction analyses. Along with the inhibition of alpha-SMA expression by bFGF, the RMF and RMFL cells also showed different activated expression of extracellular signal-regulated kinase 1/2, suggesting the involvement of extracellular signal-regulated kinase 1/2 activation in the down-regulation of alpha-SMA expression in myofibroblasts. Furthermore, an in vivo study demonstrated that bFGF administration markedly decreases the area that is positive for alpha-SMA expression in the treated wounds after day 18. In contrast, bFGF administration significantly increased the number of terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL) staining and alpha-SMA-positive cells at days 10 and 14, and reduced the double-positive cells rapidly after day 18. Collectively, the current investigation identified bFGF as a potent stimulator for the reduction of the myofibroblastic area in vivo, presumably because of its effects on the down-regulation of alpha-SMA expression as well as rapid induction of apoptosis in myofibroblasts.

Significance of lymphatic invasion and proliferation on regional lymph node metastasis in renal cell carcinoma.

We studied the associations of lymphatic invasion and lymphatic vessel density around tumors with lymph node (LN) status in renal cell carcinoma (RCC) by immunohistochemical analysis using D2-40 antibody as a lymphatic marker. Surgically removed specimens from 76 cases with RCC, including 16 cases with LN metastasis, were used. Lymphatic vessel density around the tumor increased compared with normal kidneys but was not significant by LN status. Tumor size, tumor cell types, patterns of tumor growth, nuclear grade of tumor cells, venous invasion, lymphatic invasion, and primary tumor stage were predictive factors for LN metastasis. Based on multivariate regression analysis, only lymphatic invasion was an independent risk factor for LN metastasis. The immunohistochemical detection of lymphatics was useful for identifying the lymphatic invasion of RCC, and the presence of lymphatic invasion around RCC was an independent predictive factor for LN metastasis.

Significance of lymphatic invasion on regional lymph node metastasis in early gastric cancer using LYVE-1 immunohistochemical analysis.

It has been reported that lymphatic invasion is a predictor for lymph node metastasis in early gastric cancer (EGC); however, it has been impossible to differentiate between lymphatic invasion and blood vessel invasion using current staining techniques. We studied the significance of lymphatic invasion on regional lymph node metastasis in EGC by using human lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1) antibody, specific to lymphatic vessels, and von Willebrand factor (vWF) antibody, specific to the blood vessels, to clearly distinguish these vascular tissues.EGC tissues were obtained from 66 node-positive and 66 node-negative subjects and were matched by age and sex. These tissues were immunostained with antibodies against LYVE-1 and vWF. Multivariate logistic regression analysis demonstrated that lymphatic invasion was a significant independent predictor for regional lymph node metastasis (odds ratio, 4.667; P = .0094), whereas blood vessel invasion was not. Thus, lymphatic invasion identified by LYVE-1 antibody could predict the existence of regional lymph node metastasis in EGC.

Significance of anatomical properties of myocardial bridge on atherosclerosis evolution in the left anterior descending coronary artery.

Myocardial bridge (MB) is frequently detected in the left anterior descending coronary artery (LAD), and LAD intima under MB is significantly spared from atherosclerotic evolution. Significance of anatomical features of MB on the extent of atherosclerosis of LAD was histomorphometrically investigated. Full-length 200 LADs with MB and 100 control LADs without MB were cross-sectioned at 5 mm intervals, and atherosclerosis ratio and intimal lesion types were evaluated. In cases with MB located within 5 cm from the left coronary ostium, atherosclerosis ratio in the proximal part of LAD was significantly lower than in control group, but, in cases with MB locating more than 5 cm from the ostium, atherosclerosis ratio in this part was similar to that in control cases. MB thickness was significantly correlated with its length, and the longer the MB the more proximally it tended to be located in LAD. Atherosclerosis ratio under MB was lower in cases with thick or long MBs than in cases with thinner or shorter MBs. In addition, intimal lesion in segments proximal to MB tended to be eccentric. Our results suggest that these anatomical properties of MB are the critical modulators for atherosclerosis evolution in the entire course of LAD.

[Inguinal lymph node metastasis of bladder carcinoma ten years after cystourethrectomy: a case report].

A 79-year-old man had undergone radical cystourethrectomy for bladder carcinoma in January, 1989. Pathological report was Small cell carcinoma (SCC) >> transitional cell carcinoma (TCC), G2 > G3, pT4 (prostate), ew (-). Ten years later, follow-up computed tomography (CT) revealed swollen left inguinal lymph node in October 1998 and lymph node dissection was performed in January, 2000. The pathological report showed TCC, G2. Left inguinal lymph node metastases appeared again in January, 2001. Chest X-ray films showed multiple lung metastases in March, 2001. Three couses of MVAC (methotrexate, vinblastine, doxorubicin cisplatinum) chemotherapy had been performed since September, 2001 but were in effective. Papillary tumor was observed at external urethral meatus in September, 2002 and the biopsied specimens showed TCC, G1 > G2, pathologically. Finally he died of respiratory insufficiency in January, 2003. It is suggested that the recurrent TCC tumor in the urethral remnants might metastasize into the inguinal lymph nodes.

Role of macrophage and smooth muscle cell apoptosis in association with oxidized low-density lipoprotein in the atherosclerotic development.

To examine the role of the apoptosis of macrophages and smooth muscle cells in the development of atherosclerosis, human aortic tissues with intimal lesions were immunostained with antibodies against terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL), single-stranded DNA (clone F7-26), and active caspase-3. Apoptotic cells were detected in the intima using both TUNEL and single-stranded DNA, however, the latter method was the more sensitive one for detecting apoptotic cells in the early stages of atherosclerosis. The number of apoptotic cells increased as the disease progressed. It implies that the apoptosis of intimal cells is involved in the formation of atherosclerotic lesions. In addition, quantitative analyses of the cell types undergoing apoptosis using double-immunostaining revealed that the susceptibility of macrophages and smooth muscle cells to apoptosis was greater specifically in atheroma than in the other atherosclerotic lesions, and macrophages were more susceptible to apoptosis than smooth muscle cells. The frequency and spatial distribution of oxidized low-density lipoprotein (oxLDL) (FOH1a/DLH3)-positive cells were examined by immunohistochemistry, and the results resembled those of apoptotic cells. The number of oxLDL-positive cells in the intima significantly correlated with the susceptibility of smooth muscle cells, but not with that of macrophages, to apoptosis. These results suggest that oxLDL affects the apoptosis of smooth muscle cells during the atherosclerotic development.

[Bladder carcinoma metastases presenting as intraperitoneal dissemination without local recurrence: a case report].

A 76-year-old woman presented with gross hematuria and was referred to our OPD. Cystoscopy showed broad-based papillary tumors on the left bladder wall. TUR-BT was performed and pathological diagnosis was grade 3 transitional cell carcinoma of pT1a. Although no intravesical tumor recurrence had been observed, a solid palpable mass was noted in the lower abdomen nine months after TUR-BT, and computed tomography suggested a large ovarian tumor. Subsequently performed was the operation at Gynecology, which revealed a large tumor involving the whole major omentum. Frozen sections of the tumor were diagnosed as transitional cell carcinoma metastases of the bladder cancer, and the final pathological report was the same. Although receiving 4 courses of M-VAC systemic chemotherapy after the operation, she died 14 months later. Autopsy disclosed intraperitoneal cancer dissemination and metastases without any intravesical nor left perivesical tumor recurrence, and it was suggested that the bladder tumor metastases occurred not by direct invasion but by either lymphatic or vascular mechanism in this case.

Distribution of type V secretory phospholipase A2 expression in human hepatocytes damaged by liver disease.

BACKGROUND AND AIM: Type V secretory phospholipase A2 (sPLA2-V) is a key enzyme in the arachidonate cascade. However, the distribution of sPLA2-V in human liver has not yet been investigated. In this study, the significance of sPLA2-V expression in human hepatocytes damaged by liver disease was investigated. METHODS: Samples of liver tissue from patients with chronic hepatitis B and C, hepatitis virus-related liver cirrhosis, and congestive hepatocyte injury were immunostained with antibodies against sPLA2-V, cyclooxygenase (COX)-2, hepatitis viral antigens, transforming growth factor (TGF)-beta1, interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha. RESULTS: In chronic hepatitis patients, sPLA2-V-positive hepatocytes were scattered in the liver lobules, while cyclooxygenase-2, IL-1beta, and TNF-alpha were diffusely expressed. Hepatocytes around necroinflammatory lesions were strongly positive for sPLA2-V. Some sPLA2-V-positive hepatocytes were also positive for viral antigens. TGF-beta1 was expressed only in fibrotic lesions. The pattern of distribution of these proteins in liver cirrhosis patients was similar to that in chronic hepatitis patients, but sPLA2-V expression tended to be more intense than in chronic hepatitis. In the congestive liver, sPLA2-V, COX-2, and the two cytokines were diffusely expressed in surviving hepatocytes. CONCLUSIONS: sPLA2-V expression in hepatocytes is induced by viral infection, fibrosis, and circulatory disturbance. Immunostaining using sPLA2-V antibody is useful for the detection of injured hepatocytes in patients with liver diseases.

Histiocytic sarcoma with fatal duodenal ulcers.

Histiocytic sarcoma is an uncommon neoplasm of mature histiocytes with very poor outcome. We report an autopsy case of a true histiocytic sarcoma with characteristic symptoms of so-called "malignant histiocytosis of the intestine". The liver and spleen were enlarged, with remarkable tumor cell infiltration in the hepatic sinusoids and splenic sinuses. Tumor cells aggregated to form sporadic nodular lesions in the liver, which often showed coagulative necrosis. Infarcted lesions also occurred at the splenic subcapsular area. In addition, tumor cell infiltration was noted in the sinuses of bone marrow and lymph node. Tumor cells often demonstrated moderate pleomorphism with multinucleated giant cells. They were positive for CD68 and negative for T- and B-cell lineage markers, megakaryocytic markers, and CD30. Various examinations were done to rule out infection-associated hemophagocytic syndrome, and the absence of infectious diseases was revealed. Thus, the diagnosis of histiocytic sarcoma was made. Apart from these lesions, multiple ulcerations, some with fatal perforation, were found in the esophagus and duodenum. They showed only non-specific inflammatory changes without tumor cell involvement. The ulcers probably derived from ischemic condition through an embolic process caused by tumor cell infiltration elsewhere in the blood vessels at the periphery of the ulcers.

Immunohistochemical detection of human small lymphatic vessels under normal and pathological conditions using the LYVE-1 antibody.

The spread of tumor cells via lymphatic vessels to the lymph nodes is an important indicator of malignancy. However, previous markers used to identify lymphatic endothelium gave ambiguous results in immunohistochemical analyses with paraffin-embedded tissues. In this study, we attempted to prepare a polyclonal antibody against human lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1) for detecting lymphatic vessels using immunohistochemistry. The antibody was raised against a region near the transmembrane anchor of LYVE-1 in New Zealand white rabbits. Immunostainings with anti-LYVE-1 and von Willebrand factor antibodies were performed in various normal and pathological tissues. LYVE-1 expression was confined to the endothelial surface of lymphatic vessels but was not found in the endothelium of blood vessels, which were positive for von Willebrand factor. Our LYVE-1 polyclonal antibody was useful for the identification of small lymphatic vessels in normal human tissues. In addition, the immunostaining enabled us to distinguish lymphatic invasion by malignant tumor cells from blood vessel invasion using paraffin-embedded sections. In conclusion, our polyclonal antibody against the transmembrane anchor of the peptide can be used to detect human lymphatic vessels under various conditions.

Techniques for treating aberrant arteries during resection of pulmonary sequestration by video-assisted thoracic surgery: report of two cases.

We describe the techniques we used for treating aberrant arteries during resection of pulmonary sequestration by video-assisted thoracic surgery (VATS) in two patients. In patient 1, the aberrant artery was transected after securing six rows of staples with a knifeless vascular endostapler. In patient 2, the aberrant artery was cut after ligation with special forceps that designed by one of us (S.K.) to push a knot, tied outside the body, into the thoracic cavity, then ligate the suture. Left lower lobectomy and right basal segmentectomy were both successfully performed by these methods. These two cases are reported to show that VATS lobectomy is a feasible and minimally invasive technique of treating pulmonary sequestration and other diseases of the lung.