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Reduced fibrinolytic activity is associated with adverse cardiovascular events. Recently, it has been shown that angiotensin (Ang) IV receptor (AT4R) regulated fibrinolyasis via plasminogen activator inhibitor-1 (PAI-1) activation. However, it remains unknown how much this Ang IV-AT4R signaling affects the neointimal formation in the reparative process after vascular injury. To investigate if AngIV would inhibit fibrinolysis and promote neointimal formation, we evaluated the degree of proliferation of vascular smooth muscle cells and compared with the role of AT1R using AT1R and AT4R knockout mice ( AT1R − / − and AT4R − / − , respectively). [Methods and Results] AT1R − / − and AT4R − / − showed normal hemostasis patterns including bleeding time if compared with wild type mice (C57BL6/J; WT). To assess fibrinolysis in an acute thrombosis model, renal glomerular fibrin deposition was counted after lipopolysaccharide injection (0.2mg/kg, i.v., n=5). Fibrin deposition was reduced in AT4R − / − as compared to WT and AT1R − / − at 4 hrs (12±3% of total glomeruli, 27±4% and 25±4%, p=0.012 vs. WT). The levels of plasma active PAI-1 antigen measured by ELISA were also reduced in the order of AT4R − / − , AT1R − / − and WT (4.2±0.8*, 8.0±1 and 14±3 ng/mL, *pAT1R −/− , significant hyperplasia of adventitial tissues and neointimal formation were observed after vascular injury, while those of AT4R −/− were minimally affected (n=6 each) in accordance with greater lumen diameters. In this model, genetic ablation of AT4R, but not AT1R, showed beneficial effects in vascular remodeling. [Conclusions] In murine vascular injury model, disruption of AngIV-AT4R signaling leads to accelerated fibrinolysis and decreased neointimal formation, suggesting that AT4R is a novel therapeutic target against cardiovascular disease. Morphometric Analyses of the Sections from Murine Carotid Arteries

The expression of endothelin-1 (ET-1), which has been proposed to have a potential autocrine/paracrine role, varies during the menstrual cycle, and therefore, ET-1 may be involved in the cyclic change of the human endometrium. However, neither the synthesis nor the degradation of ET-1 in the endometrium has been determined in detail. We investigated endothelin-converting enzyme-1 (ECE-1), which converts big-ET-1 to active ET-1, and neutral endopeptidase (NEP), which cleaves and inactivates ET-1 in human endometrium in vivo and in vitro. Western blot analysis demonstrated that the change in the expression of ECE-1 during the menstrual cycle differed from that of NEP in the endometrium. ECE-1 was expressed by endometrial epithelial cells, whereas NEP was predominantly expressed by stromal cells in vivo and in vitro. In conclusion, our results suggest that spacio-temporal expression of two endopeptidases, ECE-1 and NEP, involved in the synthesis and degradation of ET-1, might regulate ET-1 action in human endometrium.

A 39-year-old Japanese woman noticed a right breast tumor in July 2004. Mammography (MMG) demonstrated an oval tumor without calcification. Dynamic Magnetic Resonance Imaging (D-MRI) demonstrated a high-intensity mass on T2-weighted images, showing mild enhancement during the arterial phase and persistent enhancement during the arterial late phase. Core needle biopsy revealed papillary carcinoma suggestive of Intracystic Papillary Carcinoma (IPC). Auchincloss operation was performed following a partial mastectomy, as the surgical margin after partial mastectomy was positive for carcinoma. Histopathologic mapping of her right breast revealed wide and extensive intraductal spread of DCIS around the IPC. IPC was originally reported to be a localized non-invasive mammary carcinoma. But approximately, half of IPC cases are associated with invasive carcinoma or DCIS beyond the tumor. Careful selection of operative procedure is needed after localized non-invasive IPC or IPC associated with DCIS around the main tumor or invasive carcinoma is diagnosed.

To determine the value of peritoneal washing cytology (CY) in determining resectability of pancreatic cancer.CY has been used widely in the diagnosis and staging of several cancers. However, its predictive value in identifying potentially resectable pancreatic cancer is undetermined.Peritoneal washing samples were collected from 233 patients with pancreatic cancer between June 1991 and August 2006. A total of 157 patients had resectable and 76 had unresectable lesions. Correlations between CY status and clinicopathologic parameters with overall survival rates were analyzed.Malignant cells were identified in samples from 21 patients (13.4%) with resectable tumors and 27 patients (35.5%) with unresectable tumors. CY+ was more frequent in large tumors (or =2 cm) than small tumors (2 cm; P = 0.034). CY status did not correlate with any other clinicopathologic parameter. The overall survival of CY+ patients was worse than that of CY- patients (P = 0.047). Median survival following resection was 13.6 months for CY+ patients and 13.5 months for CY- patients. Among the patients who had unresectable lesions, median survival time was 5.9 months for CY+ and 6.1 months for CY- patients. However, among CY+ patients, those who underwent resection lived longer than those who did not (P = 0.019).Cytologic status has little predictive value for survival, and patients whose pancreatic cancer would otherwise be considered resectable should not be denied curative resection solely because they are CY+.

Intraductal papillary mucinous neoplasms (IPMNs) are increasingly recognized, but it is very difficult to evaluate accurately the malignancy of these neoplasms by modern imaging. We reviewed our experience in order to elucidate predictors of tumor malignancy, invasiveness, and outcome.The clinicopathological features and surgical outcomes of 57 patients with IPMNs who underwent surgery in Nagoya University Hospital were analyzed.The histological diagnosis was adenoma in 40, borderline in 1, carcinoma in situ (CIS) in 7, and invasive carcinoma in 9 patients. Patients with invasive carcinomas had significantly shorter survival rates than patients with benign IPMNs or CIS (p0.0001). Multivariate analyses revealed that the main duct or the combined type was significantly predictive of malignancy, and both main duct or combined type and diabetes mellitus were associated significantly with invasive carcinoma.IPMNs generally grow slowly, but have a malignant potential that warrants radical surgical treatment when the tumor component invades the parenchyma. Our results suggest that the above factors should be considered in surgical management. The main duct type of IPMN or IPMN with mural nodules is potentially malignant or invasive. Therefore, radical operative management is indicated in these IPMNs.

Malignant peripheral nerve sheath tumor (MPNST) can be difficult to diagnose because it lacks specific immunohistochemical markers. S-100, which is a useful marker of MPNST, has limited diagnostic utility. Recent studies suggest that nestin, which is an intermediate filament protein, is expressed in neuroectodermal stem cells. The diagnostic utility of immunostains for nestin and three other neural markers (S-100, CD56 and protein gene product 9.5 (PGP 9.5)) were evaluated in 35 cases of MPNST and in other spindle cell tumors. All MPNST cases were strongly positive for nestin and had cytoplasmic staining. Stains for S-100, CD56, and PGP 9.5 were positive in fewer cases (17/35, 11/35, and 29/35 cases, respectively), and had less extensive staining. Nestin was negative in 10/10 leiomyomas, and weak nestin expression was seen in 10/10 schwannomas, 3/10 neurofibromas, 2/8 synovial sarcomas, 2/10 liposarcomas, 4/7 carcinosarcomas and 3/7 malignant fibrous histiocytomas. In contrast, strong nestin positivity was seen in 10/10 rhabdomyosarcomas, 15/19 leiomyosarcomas, and 9/9 desmoplastic melanomas. Nestin is more sensitive for MPNST than other neural markers and immunostains for nestin in combination with other markers could be useful in the diagnosis of MPNST.

Background/Aims We examined whether phosphatidylcholine inhibited growth of hepatic cancer, as previously shown for menaquinone-4 (vitamin K2). Methods Growth inhibitions by phosphatidylcholine and/or menaquinone-4 and apoptosis induction by phosphatidylcholine were evaluated in vitro using human hepatic cancer cell lines (Hep-3B, Hep-G2, HuH-7, and Alexander). Effects of these agents were then investigated in male Sprague–Dawley rats against hepatocarcinogenesis induced by diethylnitrosamine plus phenobarbital. All rats were killed to examine livers to evaluate inhibitory potential macroscopically and immunohistochemically using an antibody against the marker of carcinogenesis, glutathione S -transferase and apoptotic induction by phosphatidylcholine using TUNEL staining. Blood samples were obtained by cardiac puncture. Results In vitro , phosphatidylcholine and menaquinone-4 each inhibited cancer cell growth and phosphatidylcholine induced apoptosis dose-dependently. Moreover, exposure to both synergistically inhibited growth in Hep-3B. In vivo , diets containing phosphatidylcholine with or without menaquinone-4 significantly reduced the number of macroscopic hepatic tumor nodules and the extent of abnormally immunoreactive foci conserving hepatic function on serum examinations compared with controls given only the carcinogens. Moreover, phosphatidylcholine supplementation induced apoptosis on TUNEL staining of liver sections. Conclusions Given together, phosphatidylcholine and menaquinone-4 may exhibit synergy against hepatocarcinogenesis conserving hepatic function that could benefit patients at high risk for hepatocellular carcinoma.

Loss of E-cadherin triggers peritoneal dissemination, leading to an adverse prognosis for most patients with epithelial ovarian carcinoma (EOC). Because TWIST mainly regulates the epithelial-to-mesenchymal transition and is one of the E-cadherin repressors, we investigated the possibility that TWIST expression affects peritoneal metastasis of EOC using siRNA technique. In the present study, we showed a correlation between TWIST expression and EOC cellular morphology. Furthermore, we demonstrated that the suppression of TWIST expression in EOC cells (HEY) alters the cellular morphology from a fibroblastic and motile phenotype to an epithelial phenotype, and inhibits the adhesion of these cells to mesothelial monolayers. To investigate the mechanism by which down-regulation of TWIST leads to inhibition of adhesion to mesothelial cells (MCs), expression of adhesion molecules (CD29, CD44 and CD54) were observed. Moreover, matrix metalloproteinase 2 and membrane type 1 matrix metalloproteinase, important markers associated with invasive and metastatic potential, were remarkably reduced. This findings suggests that reduced expression of TWIST suppresses the multistep process of peritoneal dissemination (detachment from the primary lesion, adhesion to MCs and invasion of MCs) and may be a potential therapeutic target for the treatment of this carcinoma.

金沢大学大学院医学系研究科がん細胞学, Tumor-stromal interactions are important for the progression of malignant tumors. The purpose of the present study was to examine interactions of cholangiocarcinoma (CC) cells and stromal fibroblasts with respect to stromal-derived factor-1 (SDF-1) and transforming growth factor (TGF)-β1. Two cell lines of CC (HuCCT-1 and CCKS-1) and WI-38 fibroblast cell line were used for cell culture, and 12 CC tissue specimens for immunohistochemical studies. Invasion of CC cells was increased significantly by the supernatant from fibroblast cultures, but not by the supernatant from fibroblasts cocultured with CC cells. Expression of SDF-1 in cultured fibroblasts was downregulated by TGF-β1 treatment, and coculture with CC cells and anti-TGF-β1 neutralizing antibody restored the decreased SDF-1 expression, suggesting that TGF-β1 secreted from CC cells might have reduced the expression of SDF-1 by fibroblasts and might have reduced the increased invasion of CC cells induced by the supernatant from fibroblasts. Immunohistochemical expression of TGF-β1 in CC cells was focal or negative and that of SDF-1 was evident in stromal fibroblasts at the invasive front of CC. In conclusion, local mutual influence of TGF-β1 secreted from carcinoma cells and SDF-1 expressed by stromal fibroblasts may be involved in invasion of CC cells. © 2006 Japanese Society of Pathology.

Malignant lymphoma is a heterogeneous category embracing three major types of lymphoid neoplasms: B cell neoplasms, T and NK cell neoplasms, and Hodgkin lymphoma. Within each type, distinct disease entities are defined based on a combination of morphology, immunophenotype, genetic features and clinical syndromes, the emphasis on which represents a new paradigm in the lymphoma classification of the World Health Organization (WHO). These lymphoma entities often have distinctive cytogenetic abnormalities, usually involving translocations that place a potential cellular oncogene under the influence of the immunoglobulin in some low-grade B-cell lymphomas. Both pathologists and oncologists are now concerned with better understanding each disease entity and its spectrum of morphology, genetic events, and clinical behaviors. Over the last decade, significant progress has been made in the molecular characterizations of mantle cell lymphoma, anaplastic large cell lymphoma, and marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT), which have not only provided insights into the pathogenesis of lymphomas, but also valuable data that could lead to therapies based on their clinical behavior.

Angiotensin II, a main effector peptide in the renin-angiotensin system, acts as a growth-promoting and angiogenic factor via type 1 angiotensin II receptors (AT(1)R). We have recently demonstrated that angiotensin II enhanced tumour cell invasion and vascular endothelial growth factor (VEGF) secretion via AT1R in ovarian cancer cell lines in vitro. The aim of the present study was to determine whether AT1R expression in ovarian cancer is correlated with clinicopathological parameters, angiogenic factors and patient survival. Immunohistochemical staining for AT1R, VEGF, CD34 and proliferating cell nuclear antigen (PCNA) were analysed in ovarian cancer tissues (n = 67). Intratumour microvessel density (MVD) was analysed by counting the CD34-positive endothelial cells. Type 1 angiotensin II receptors were expressed in 85% of the cases examined, of which 55% were strongly positive. Type 1 angiotensin II receptors expression was positively correlated with VEGF expression intensity and MVD, but not with histological subtype, grade, FIGO stage or PCNA labelling index. In patients who had positive staining for AT1R, the overall survival and progression-free survival were significantly poor (P = 0.041 and 0.017, respectively) as compared to those in patients who had negative staining for AT1R, although VEGF, but not AT1R, was an independent prognostic factor on multivariate analysis. These results demonstrated that AT1R correlated with tumour angiogenesis and poor patient outcome in ovarian cancer, suggesting its clinical potential for a novel molecular target in strategies for ovarian cancer treatment.

Hemangiomas arising from tendon and tendon sheath are rarely reported, and may be confused with other lesions of tendons. In this case report, a 19-year-old woman was diagnosed with a cavernous hemangioma originating in the left gracilis tendon and tendon sheath. When her symptoms initially developed, MRI did not delineate the lesion due to the relatively small size of the tumor. Nine years after the onset of the patient's complaints of knee pain and swelling, the tumor was adequately diagnosed on physical examination and MRI, and was resected with complete relief of symptoms.

The possible role of calcineurin in cardiac hypertrophy induced by calmodulin (CaM) overexpression in the heart was investigated. CaM transgenic (CaM-TG) mice developed marked cardiac hypertrophy and exhibited up-regulation of atrial natriuretic factor (ANF) and beta-myosin heavy chain gene expression in the heart during the first 2 weeks after birth. The activity of calcineurin in the heart was also significantly increased in CaM-TG mice compared with wild-type littermates. Treatment of CaM-TG mice with the calcineurin inhibitor FK506 (1mg/kg per day) prevented the increase in the heart-to-body weight ratio as well as that in cardiomyocyte width. FK506 also inhibited the induction of fetal-type cardiac gene expression in CaM-TG mice. Overexpression of CaM in cultured rat cardiomyocytes activated the ANF gene promoter in a manner sensitive to FK506. Activation of a calcineurin-dependent pathway thus contributes to the development of cardiac hypertrophy induced by CaM overexpression in the heart.

Objective To assess the expression and cellular distribution of angiotensin II (Ang II), angiotensin type 1 receptor (AT1R), angiotensin-converting enzyme (ACE), aminopeptidase A (APA), adipocyte-derived leucine aminopeptidase (A-LAP), and vascular endothelial growth factor (VEGF) in human ovarian tissue during the menstrual cycle. Design Ovarian tissues (n = 52) and corpora lutea (n = 34) were obtained from patients undergoing hysterectomy/oophorectomy, and tissue sections were immunostained for each antigen. Setting University hospital. Patient(s) Patients undergoing hysterectomy or oophorectomy for benign conditions. Intervention(s) Immunostaining of tissue sections using antibodies to each antigen. Main Outcome Measure(s) Microscopic evaluation to assess the presence, distribution, and cellular localization. Result(s) The luteal tissue is the major site of Ang II, ACE, AT1R, and VEGF, with highest staining intensity found during the midluteal phase and at pregnancy. The AT1R was found in theca cells. The APA was strongly immunolocalized in pericytes. Immunolocalization of AT1R was almost similar to that of VEGF including oocytes in the primordial and intermediate follicles. Conclusion(s) The expression and distinct pattern of the cellular localization of Ang II and its related proteins in human ovarian tissue during folliculogenesis and in the luteal tissue suggest their roles in the growth and differentiation of theca, granulose, and luteal cells.

An oncolytic herpes simplex virus type 1 mutant (HF10) has been isolated and evaluated for antitumor efficacy in a syngeneic immunocompetent mouse model, where it was effective against cancer and conferred resistance to rechallenge with tumor cells in all surviving mice. Several studies have shown that HF10 is effective and safe for use against localized or peritoneally disseminated nonneuronal malignant tumors in animals.A pilot study using HF10 was initiated in six patients with cutaneous or subcutaneous metastases from breast cancer. For each patient, .5 mL of HF10 suspension containing various viral doses was injected into one nodule; .5 mL of sterile saline was injected into another. All patients were monitored for local and systemic adverse effects. Nodules were excised 14 days after injection for histopathologic studies.All patients tolerated the intratumoral injection of HF10. No adverse effects occurred, and histopathological evaluation revealed 30% to 100% cancer cell death.This pilot study found HF10 to be safe and effective against metastatic breast cancer.

Apocrine carcinoma is an extremely rare malignant neoplasm that occurs most frequently in the axilla. Although it usually shows an indolent clinical course, it often metastasizes to regional lymph nodes and sometimes to lungs or bones. However, a literature search did not reveal any report describing the detailed clinical course of brain metastases from apocrine carcinoma. We report a case of a 54-year-old male who suffered from multiple brain metastases from apocrine carcinoma that had originated in the scalp 6 years before. The brain metastases appeared in spite of several regimens of chemotherapy for lung metastases for two years. The tumor in the right frontal lobe was successfully operated. However, the small tumor in the right occipital lobe was not cured by gamma knife surgery, and eventually required second operation. The operation had contributed to his neurologically independent life for about one year until he died for gradual progression of lung metastases. To our knowledge this is the first reported case of metastatic brain tumor from apocrine carcinoma.

The authors have recently defined the clinopathological entity of a mucin-producing bile duct tumor (MPBT), and divided MPBT into two distinct subtypes: ‘columnar-type’ and ‘cuboidal-type’ MPBT. Mucin core protein 6 (MUC6), which is present in normal pyloric glands, had higher expression levels in cuboidal-type tumors than in columnar-type tumors. In the pyloric glands, a carbohydrate antigen detected by monoclonal antibody HIK1083 (CA/HIK1083) is also expressed. In order to evaluate the coexpression pattern of MUC6 and CA/HIK1083 in MPBT, expression profiles were evaluated in 38 surgically excised mucin-producing bile duct carcinomas (MPBC; cuboidal-type, n = 15; columnar-type, n = 23), using immunohistochemistry. The staining rate was graded as follows: –

Although size and mitotic counts have been reliable predictors of clinical outcome, identification of gastrointestinal stromal tumor (GIST) of the stomach with a metastatic potential through hematoxylin and eosin staining is not always accurate. Telomerase activity, often detected in malignant tumors, may have a role as a marker for high-grade malignancy.Immunostaining with antibodies against KIT protein, CD34, and other molecules that are relevant for evaluation of cell differentiation and proliferation was performed for 36 primary gastric submucosal tumors to confirm the diagnosis of GIST. DNA was extracted from the surgically resected specimens of 24 of 36 patients, and c-kit mutation was analyzed by direct sequencing after PCR amplification of the exon 11. Telomerase activity was quantitatively evaluated for all 36 patients using fluorescence-based telomeric amplification assay protocol (TRAP) analysis.c-kit mutation was observed in 58% of the patients evaluated. Telomerase activity was detected in 14 specimens (39%), but not in the specimens without c-kit mutation. All 5 patients who suffered from metastatic or recurrent disease exhibited c-kit mutation and a high level of telomerase activity.Measurement of telomerase activity, along with c-kit mutation analysis, is useful for identifying GIST with a potential for malignant behavior.

Surgical pathologists often encounter hydropic villi in products of conception at the first trimester and must determine whether the villi represent complete hydatidiform mole (CM), partial hydatidiform mole (PM), or hydropic abortion (HA). The distinction between these is important for determining the appropriate treatment of patients. This study assessed interobserver and intraobserver variability in the histologic diagnosis of hydatidiform mole among 5 placental pathologists. To evaluate interobserver variability, one representative slide from each of 50 mixed cases of PM, CM, and HA of the first trimester were circulated among 5 placental pathologists. All pathologists used the same histologic criteria by Szulman and Surti. For the second round, the same cases were submitted with DNA ploidy data. For the third round, the slides were recoded and distributed to assess intraobserver agreement. Kappa (kappa) value was calculated for the interobserver agreement in the first and second rounds. There was agreement among 4 or 5 pathologists for only 30 of 50 cases in the first round. There were problems in differentiating between PM and HA in most of the remaining 20 cases. The kappa values varied from poor (kappa = -0.104) to excellent (kappa = 0.761) in the first round. In the second round, there was agreement in 39 of 50 cases and the level of agreement remarkably increased, ranging from fair to good (kappa = 0.552) to excellent (kappa = 0.851). The number of discrepant cases, PM versus HA, was reduced to 4. In 7 cases, there were difficulties in distinguishing CM from HA. The intraobserver agreement ranged from 50% to 90%. Poor interobserver agreement was demonstrated when histology alone was used for diagnosis. Discordance was most frequently seen in PM versus HA and resulted from difficulty in evaluating trophoblastic hyperplasia. Polar trophoblastic growth seen in HA could also be observed in PM. The addition of ploidy data resulted in a significant improvement in concordance. Ploidy study is useful in equivocal cases. Significant interobserver and intraobserver variability was observed even among placental pathologists. New histologic criteria adaptable to differentiation of early lesions are needed.

Atypical teratoid/rhabdoid tumors (AT/RTs) are aggressive malignant tumors found in infants and young children. The tumor is characterized by the presence of a rhabdoid cell component in all cases, but the histological origin is still unclear. Recently, germline mutation of the hSNF5/INI1 gene has been reported in association with AT/RTs. The authors report a rare case of an intraocular AT/RT followed by a fourth ventricular tumor. The results of immunohistochemical studies of the surgical specimens revealed the presence of an AT/RT and from this finding the neural origin was inferred. A novel missense mutation of the hSNF5/INI1 gene was demonstrated by DNA analysis. High-dose chemotherapy with stem cell rescue was effective in treating this patient. The immunohistochemical relationship between rhabdoid cells and the neurogenic zone, which has not been described in AT/RTs, is of great interest in view of the nature of rhabdoid cells.

Background Infectious mononucleosis owing to primary Epstein-Barr virus (EBV) infection sometimes causes hepatitis, which is usually self-limiting with mildly elevated transaminases, but can rarely develop into severe hepatitis with jaundice. Objective To clarify the pathogenesis of severe hepatitis by primary EBV infection. Methods We experienced four cases of severe hepatitis with jaundice caused by primary EBV infection. These cases were analyzed virologically and histologically, and compared with infectious mononucleosis patients without jaundice. Results and discussion Using real-time PCR, more EBV-DNA was detected in peripheral blood from patients with severe hepatitis, as compared to those without jaundice. Furthermore, CD3 + , CD4 + or CD8 + cells contained more EBV DNA than did other cell populations, indicating that in severe hepatitis, T cells harbor most of the EBV. By contrast, mainly B cells were infected in infectious mononucleosis patients without jaundice. The liver was biopsied in three of the four cases. An in situ hybridization study showed that EBV infected lymphocytes, not hepatocytes. In addition, in one patient, it was confirmed that the infected lymphocytes were CD8 + T cells. These results suggest that a large EBV burden and T cell infection may play major roles in the mechanism of severe hepatitis caused by primary EB virus infection.

Intraportal endovascular ultrasound (IPEUS) has been reported to be the most precise diagnostic procedure for the accurate diagnosis of portal vein/superior mesenteric vein (PV/SMV) invasion in patients with pancreatic cancer. In this study, we evaluated the clinical significance of the length of PV/SMV invasion measured by IPEUS METHODOLOGY: Twenty-six consecutive patients who underwent the pancreatic resection and IPEUS using an autopull-back device between January, 1997 and September, 2000 were retrospectively evaluated. The length of PV/SMV invasion was measured by reviewing the videotapes recorded during the operation. Clinicopathological data and survival were analyzed.The percentage of PV/SMV invasion was 46%, all of which were treated by PV/SMV resection. The cases without PV/SMV invasion showed significantly longer survival rate. The cases withor = 18mm PV/SMV invasion, however, achieved a comparable 2-year survival rate of 28% whereas no patient with18mm PV/SMV invasion survived more than 18 months after the resection.Involvement of the PV/SMV by pancreatic carcinoma seems to be related to the extent of the disease and the PV/SMV involvement18mm is associated with a poor prognosis due to high rate of tumor positive margin even with radical operation.

A 46-year-old man was referred to our hospital for the treatment of lung cancer. Computed tomography showed a well-defined tumor mass that was 50×45 mm in size and contained a trabecular pattern of calcification. Since he was diagnosed as having a primary lung adenocarcinoma (clinical stage IB), a left upper lobectomy with mediastinal lymph node dissection was performed. Histologically, the tumor was a poorly differentiated adenocarcinoma with rich fibrous stroma, in which there were island-shaped bone formation lesions. An immunohistochemical examination showed the expression of bone morphogenic protein-2 within tumor cells, which induce and stimulate bone formation. This finding may elucidate a possible mechanism of heterotopic bone formation.

Background Small-bowel enteroscopy with the double-balloon method was developed to improve access to the small intestine. This study evaluated the usefulness of this method for the resection of small-intestinal Peutz-Jeghers polyps. Methods Two patients with Peutz-Jeghers syndrome underwent nonsurgical double-balloon enteroscopic resection of polyps throughout the small intestine. Observations Multiple polyps in the jejunum were successfully resected via the oral route, as were the polyps in the ileum via the anal route. All 18 polyps (10-60 mm in size) were resected without subsequent bleeding or perforation. Histopathologically, 3 large polyps (>30 mm diameter) were hamartomas with adenomatous components. Conclusions Double-balloon enteroscopy was safe and useful for the diagnosis and the treatment of Peutz-Jeghers polyps throughout the small intestine. Double-balloon enteroscopic polypectomy might preclude complications of Peutz-Jeghers syndrome, including intussusception, bleeding, and tumorogenesis, thereby obviating the need for multiple laparotomies.

The epidermal growth factor receptor (EGFR) gene has recently been reported to be mutated in a subset of non-small cell lung cancers (NSCLC), with the mutations being correlated with the patients' drug sensitivity to gefitinib, an EGFR kinase inhibitor. In this study, we searched for EGFR mutations in patients with lung cancer using primary tumor specimens obtained at initial surgery and examined whether their recurrent tumors showed a response to gefitinib depending on the presence of the activating mutation. Among 12 lung cancers that were treated with gefitinib after recurrence, we found that all four tumors which showed a response to gefitinib had an activating mutation in EGFR, whereas none of the remaining eight tumors had a mutation. Southern blot analysis showed that two of the four responsive tumors had the EGFR gene amplification. We also examined another 73 NSCLC specimens (47 males and 26 females; 53 adenocarcinomas and 20 non-adenocarcinomas) which were not treated with gefitinib to determine whether NSCLCs with an EGFR mutation have different clinicopathological properties and/or unique genetic alterations of the other cancer-associated genes. We found that 13 (18%) of 73 tumors had a mutation of the EGFR gene, with the most being detected in female adenocarcinomas. Comparing the alterations in KRAS and P53 with the EGFR mutation, we found that 10 tumors with the KRAS mutation did not have an EGFR mutation, suggesting that each mutation occurs exclusively during the development of lung cancer. These results suggest that the mutation analysis of the EGFR gene using the specimens obtained at surgery might be useful in selecting the appropriate treatment(s) for recurrent lung cancer patients.

The present study investigated a correlation between osseointegration in dental implants and an injectable tissue-engineered bone, using mesenchymal stem cells (MSCs) and platelet-rich plasma (PRP). Initially, the teeth in the mandible region were extracted and the healing period was 1 month. Bone defects on both sides of the mandible were prepared with a trephine bar. The defects were implanted with graft materials as follows: PRP, dog MSCs (dMSCs), and PRP, autogenous particulate cancellous bone and marrow (PCBM), and control (defect only). Two months later, the animals were evaluated by histology, and at the same time dental implants were installed. Two months later, the animals were sacrificed and nondecalcified sections were evaluated histologically and histometrically. According to the histological observations, the dMSCs/PRP group had well-formed mature bone and neovascularization, compared with the control (defect only) and PRP groups, as was the same for the PCBM group. A higher marginal bone level was observed around implants with PRP, PCBM, and dMSCs/PRP compared with the control. Furthermore, the values describing the amount of bone-implant contact (BIC) at the bone/implant interface were significantly different between the PRP, PCBM, dMSCs/PRP, and control groups. Significant differences were also found between the dMSCs/PRP and control groups in bone density. The findings of this experimental study indicate that the use of a mixture of dMSCs/PRP results in good results such as the amount of BIC and bone density comparable with that achieved by PCBM.

Adipocyte-derived leucine aminopeptidase (A-LAP, endoplasmic reticulum aminopeptidase ERAP1) is specialized to produce peptides presented on the class I major histocompatibility complex (MHC) by trimming epitopes to eight or nine residues, in addition to its enzymatic activity to degrade angiotensin II. Previously we identified placental leucine aminopeptidase (P-LAP), another member of the oxytocinase subfamily of aminopeptidases, in human uterine endometrial epithelial cells. Here we analyzed the distribution of A-LAP in human cyclic endometrium. Western blotting analysis showed that A-LAP was present in the endometrial tissue throughout the menstrual cycle. Immunohistochemical (IHC) analysis of A-LAP showed a similar distribution to that of P-LAP. A-LAP was localized predominantly in the endometrial glands and the luminal surface epithelium. However, the intracellular localization change that accompanied apocrine secretion, which was observed in P-LAP, was not shown in A-LAP. Subcellular localization of A-LAP, demonstrated by immunofluorescence, was ER in the cultured glandular epithelial cells. Our results indicate that A-LAP may fit the endometrial localization as an antigen-presenting ER aminopeptidase. Further understanding of the function(s) of A-LAP in the endometrium appear likely to yield insights into the cyclic changes during the normal endometrial cycle.

We have attempted to regenerate bone in a significant osseous defect with minimal invasiveness and good plasticity, and to provide a clinical alternative to autogenous bone grafts. Platelet-rich plasma (PRP) may enhance the formation of new bone and is nontoxic, nonimmunoreactive, and accelerates existing wound-healing pathways. We have used a combination of PRP as an autologous scaffold with in vitro-expanded mesenchymal stem cells (MSCs) to increase osteogenesis, compared with using the scaffold alone or autogenous particulate cancellous bone and marrow (PCBM). The newly formed bones were evaluated by radiography, histology, and histomorphometric analysis in the defects at 2, 4, and 8 weeks. According to the histological observations, the dog MSCs (dMSCs)/PRP group had well-formed mature bone and neovascularization compared with the control (defect only), PRP, and PCBM groups at 2 and 4 weeks. Histometrically, at 8 weeks newly formed bone areas were 18.3 +/- 4.84% (control), 29.2 +/- 5.47% (PRP), 61.4 +/- 3.38% (PCBM), and 67.3 +/- 2.06% (dMSCs/PRP). There were significant differences between the PCBM, dMSCs/PRP, and control groups. These results demonstrate that the dMSCs/PRP mixture is useful as a osteogenic bone substitute.

Infections of T cells and natural killer (NK) cells play a central role in the pathogenesis of chronic active Epstein-Barr virus (CAEBV) infection. To characterize the virologic and cytokine profiles of T cell-type and NK cell-type infection, 39 patients with CAEBV infection were analyzed. Patients with T cell-type infection had higher titers of immunoglobulin G against early and late EBV antigens, suggesting lytic cycle infection. However, the pattern of EBV gene expression was latency type II; BZLF1, which is a hallmark of lytic cycle infection, could not be detected in any patients, regardless of infection type. Patients with CAEBV infection had high concentrations of proinflammatory, T helper cell type 1, and anti-inflammatory cytokines. The cytokine profile in patients with NK cell-type infection was similar to that in patients with T cell-type infection, but the concentration of IL-13 was high in patients with NK cell-type infection. These findings should help to clarify the pathogenesis of CAEBV infection and facilitate the development of more-effective treatments.

Translational research involves application of basic scientific discoveries into clinically germane findings and, simultaneously, the generation of scientific questions based on clinical observations. At first, as basic research we investigated tissue-engineered bone regeneration using mesenchymal stem cells (MSCs) and platelet-rich plasma (PRP) in a dog mandible model. We also confirmed the correlation between osseointegration in dental implants and the injectable bone. Bone defects made with a trephine bar were implanted with graft materials as follows: PRP, dog MSCs (dMSCs) and PRP, autogenous particulate cancellous bone and marrow (PCBM), and control (defect only). Two months later, dental implants were installed. According to the histological and histomorphometric observations at 2 months after implants, the amount of bone–implant contact at the bone–implant interface was significantly different between the PRP, PCBM, dMSCs/PRP, native bone, and control groups. Significant differences were also found between the dMSCs/PRP, native bone, and control groups in bone density. These findings indicate that the use of a mixture of dMSCs/PRP will provide good results in implant treatment compared with that achieved by autogenous PCBM. We then applied this injectable tissue-engineered bone to onlay plasty in the posterior maxilla or mandible in three human patients. Injectable tissue-engineered bone was grafted and, simultaneously, 2–3 threaded titanium implants were inserted into the defect area. The results of this investigation indicated that injectable tissue-engineered bone used for the plasty area with simultaneous implant placement provided stable and predictable results in terms of implant success. We regenerated bone with minimal invasiveness and good plasticity, which could provide a clinical alternative to autogenous bone grafts. This might be a good case of translational research from basic research to clinical application.

An unusually rare case of atypical endometrial hyperplasia with clear cell change (metaplasia) spreading through almost the whole endometrium of a 37-year-old woman with infertility and abnormal bleeding is reported.

Several studies have suggested that pancreatic exocrine cells are able to differentiate into endocrine cells. REG I has been shown to be crucial for induction of ductal epithelial cells to differentiate into insulin producing cells. To date, however, there has been no study which examined the expression of REG I along with other differentiation markers in patients with chronic pancreatitis.Six cases with chronic pancreatitis and the pancreata from the age-matched autopsy cases of unrelated diseases (n=2) were included in this study and examined by immunostaining methods using antibodies against human REG I protein, trypsin, insulin, chromogranin A, cytokeratin 19 and Ki-67.In chronic pancreatitis, REG I was found in acinar and acinoductular cells and the latter were also characterized by cytokeratin 19 positivity. The acinoductular cells showed the budding of REG I-positive cells which were also trypsin- and chromogranin A-positive. MIB-1 antibody against Ki-67 was found negative in the ductal, acinoductular and islet cells reflecting low proliferation in these cells.The current study revealed that acinoductular cells characterized by dual expression of REG I and cytokeratin 19 also express insulin, chromogranin A and trypsin and therefore supports the hypothesis that transdifferentiation of fully differentiated (acinar) cells may be the main source of islet neogenesis in patients with chronic pancreatitis. However, low proliferation rate in these cells limits the regeneration capacity of the pancreatic tissue in these cases. The present results of double staining of REG I with cytokeratin 19, chromogranin A and insulin in the acinoductular cells indicate that REG I may be used as a marker for these cells.

Alopecia areata (AA) has been suspected to be an autoimmune disease, although there is no distinct evidence, we investigated the relationship between AA and autoantibodies against dense fine speckles 70 kDa (DFS70) in 111 patients with alopecia and 105 healthy controls. The sera from 59 out of 111 (53%) Japanese alopecia patients were positive for anti-nuclear antibody (ANA), as compared to the sera of 16 out of 105 (15%) healthy controls (p < 0.001). Twenty percent (22/111) of the alopecia patients were shown to be positive for the prevalence of anti-DFS70 antibodies, as compared to 8% (8/105) of the healthy controls (p < 0.01). IgG subclass analysis by ELISA showed that IgG1 and IgG2-anti-DFS70 antibodies were dominant in alopecia patients. The DFS70 gene expression in the hair structures was clearly detected in both those with and those without the anti-DFS70 antibody by RT-PCR. Immunohistochemical techniques showed that the DFS70 was localized predominantly in the outer root sheath (ORS) cells. The elevated anti-DFS70 antibodies in alopecia patients and the localization of the DFS70 in the ORS suggest that autoantibodies against the DFS70 are related to the etiology in a certain population of AA.

Glucose transporter 4 (GLUT4) is the main insulin-responsive glucose transporter in skeletal muscle and adipose tissue of human and rodent, and is translocated to the plasma membrane in response to insulin. GLUT2 is well known as the main glucose transporter in pancreatic islets and could highly regulate glucose-stimulated insulin secretion by B-cells as a glucose sensor. We confirmed the presence of GLUT4 mRNA and GLUT4 protein in pancreas in the human. Indirect immunohistochemistry showed that the pancreatic islets of human and rat were conspicuously labeled by anti-GLUT4 antibody. The presence of placental leucine aminopeptidase (P-LAP), a homologue of insulin-regulated aminopeptidase (IRAP), was also shown in the human pancreatic islet. IRAP/P-LAP is thought to be involved in glucose metabolism. This study provides the first evidence that GLUT4 is present in human and rat pancreatic islets and may suggest its specific role in glucose homeostasis in conjunction with IRAP/P-LAP.

Summary Background: Mucosal high-risk human papillomaviruses (HPVs), such as type 16, are detectable in oral cancers, especially of the oropharynx and tonsils, and there is evidence that they play a pathogenetic role in some cases. However, information is limited about their significance for cancers of the vermilion of the lip. Objective: To determine the detection rate, types and localization of HPVs in squamous cell carcinomas (SCCs) of the lip. Methods: Nested PCR for cutaneous HPVs, including epidermodysplasia verruciformis-related HPV (EV-HPV), and single PCR for mucosal HPVs, were conducted for a total of 27 SCCs and normal samples from 30 individuals. Tyramide-based in situ hybridization (ISH) was also applied. Results: Various types of HPVs were detected, particularly from normal individuals. Among the kinds of the HPV types detected in this study, half were found by PCR using a primer pair, which we newly designed. The prevalence of HPV was 5 out of 27 SCCs (ca. 18%) and 10 out of 30 normal individuals (ca. 33%). They were the entire cutaneous-group except for two, from one SCC and one normal individual. Conclusion: On the surface of the normal lip various types of mainly cutaneous-group HPVs may be present, but there does not appear to be any obvious association with SCCs developing in this site.

We recently identified two novel aminopeptidases, placental leucine aminopeptidase (P-LAP) and adipocyte-derived leucine aminopeptidase (A-LAP). Enzymatically, P-LAP degrades oxytocin, vasopressin, and angiotensin III, while A-LAP degrades angiotensin II and kallidin. In this study we investigated the expression and localization of P-LAP and A-LAP in human trophoblastic cells in the normal placenta (n = 26), gestational choriocarcinoma (n = 8), and placental site trophoblastic tumor (n = 3). On immunoblot analysis both P-LAP and A-LAP proteins were detected in normal placenta and five choriocarcinoma tissues, as well as in two choriocarcinoma cell lines. Immunohistochemical staining of normal placental tissues demonstrated that P-LAP was not only localized in villous syncytiotrophoblasts but also highly expressed in extravillous trophoblasts (EVTs) invading the decidua or maternal spiral arteries. The expression level of P-LAP on these invasive EVTs reached a maximum during the late first to second trimesters of pregnancy, and it decreased in the third trimester. Similarly, A-LAP was strongly expressed in EVTs invading the decidua or spiral arteries in the second trimester of pregnancy, while it was weakly or moderately expressed in villous cytotrophoblasts or EVTs located in the cell columns. These two aminopeptidases were more strongly expressed in all eight choriocarcinomas and three placental site trophoblastic tumors and mainly localized to the intermediate-type trophoblastic tumor cells invading the uterine myometrium or stromal vessels. In summary P-LAP and A-LAP were predominantly expressed in the invasive phenotype of EVTs during placentation, as well as in the invasive tumor cells of trophoblastic neoplasms. These results suggest the involvement of these aminopeptidases in invasiveness of both normal and malignant intermediate-type trophoblasts possibly through degradation of specific peptide substrates.

PGP9.5 is a ubiquitin hydrolase widely expressed in neuronal tissue at all stages of neuronal differentiation and has been used as a neuroendocrine marker. Recently, it has been proved that PGP9.5 expression was highly observed in squamous cell carcinoma of lung cancer, suggesting that it might be a tumor marker for squamous cell carcinoma. To better characterize its role in digestive tract cancers, we examined PGP9.5 expression retrospectively in esophageal cancers.We examined PGP9.5 expression retrospectively in 40 resected esophageal cancers (squamous cell carcinoma) and 10 gastric cancers (adenocarcinoma) using immunohistochemistry.Of 40 esophageal cancer specimens, 19 (48%) exhibited positive staining with PGP9.5 in most tumor cells, while no PGP9.5 expression was observed in any of the 10 gastric cancers.Although the precise mechanism underlying the effect of PGP9.5 on oncogenicity remains to be proven, it was confirmed that it may be a potential marker for esophageal squamous cell carcinoma.

Background Although Cushing’s syndrome arises from various neuroendocrine tumors secreting adrenocorticotropin (ACTH) ectopically, ovarian carcinoma rarely causes this syndrome. Case A 66-year-old woman presented with facial swelling and skin pigmentation. She manifested hypercortisolemia, high plasma ACTH, and lack of dexamethasone suppression. MRI showed a solid ovarian tumor and resection of the tumor led to normalization of ACTH and cortisol levels. In addition, elevated serum vasopressin (ADH) and α-fetoprotein (AFP) were found, which were also normalized after removal of tumors. Pathological diagnosis was serous adenocarcinoma with neuroendocrine and hepatoid features. Immunohistochemistry detected immunoreactivity of chromogranin A, ACTH, ADH, and AFP in tumor cells. Conclusion This is a very rare case of successful treatment of Cushing’s syndrome arising from an ovarian adenocarcinoma secreting multiple endocrine substances.

Although the presence of an intramural metastasis (IM) from esophageal cancer is associated with a poor prognosis, some patients with IM have a relatively favorable course.Clinicopathological factors including number, location, and size of IM and the distance between the primary tumor and IM were assessed in 212 patients with esophageal squamous cell carcinoma who underwent esophagectomy.Twenty-three patients (10.8%) had IM. IM size ranged from 2 to 100 mm (18+/-26), and the distance between primary lesion and IM ranged from 5 to 70 mm (27+/-18). Survival of ten patients with an IM less than 20 mm from the primary tumor was significantly longer than that of 13 patients with a more distant IM ( P=0.0184); median survival time were 2.3 and 0.7 years, respectively.A subgroup of patients with an IM less than 20 mm from the primary esophageal cancer may have a relatively favorable prognosis. When an IM is found preoperatively or in a resected specimen, measurement of the distance between the primary tumor and the IM might be useful in determination of treatment strategy and evaluation of prognosis.

We report a rare case of metachronous double cancer of the biliary tract. At age 59 years, a man had undergone a cholecystectomy and resection of the liver bed for gallbladder cancer pathologically diagnosed as papillary adenocarcinoma, in 1997. Four years later, he was admitted to our hospital with jaundice. At first, we suspected lymph node metastasis of the gallbladder cancer along the common bile duct. But abdominal computed tomography demonstrated circular wall thickness of the common bile duct, so primary bile duct cancer was strongly suspected. Thus, extended right hepatectomy and pancreaticoduodenectomy were performed after right portal vein embolization. The pathological diagnosis of the resected specimen was well-differentiated tubular adenocarcinoma, and this case was clarified to be metachronous double cancer. A review of the literature regarding double cancer of the biliary tract is presented following this case report. We showed that half of 30 cases of double cancer of the biliary tract were not associated with pancreaticobiliary maljunction, including all 6 metachronous cases.

Objective: Placental leucine aminopeptidase (P-LAP) is a cell surface aminopeptidase (oxytocinase). We cloned P-LAP cDNA and found a widespread tissue distribution of P-LAP. Since P-LAP can degrade several small peptide hormones such as oxytocin, this enzyme may affect many cellular functions of carcinoma cells as well as normal cells. This study investigated whether the expression of P-LAP correlates with clinicopathologic factors and prognosis in patients with endometrial endometrioid adenocarcinoma. Methods: Histologic sections of formalin-fixed, paraffin-embedded specimens from 99 primary endometrial carcinomas were stained for P-LAP using polyclonal P-LAP antibody. Disease-free survival and other clinicopathologic characteristics were analyzed according to the intensity of P-LAP staining. Results: Of 99 cases, 69 (69.7%) showed specific P-LAP immunostaining. We found a positive correlation between the expression of P-LAP and histological grade (p < 0.01), surgical stage of the disease (p = 0.02), myometrial invasion (p = 0.01), lymph node involvement (p < 0.01), and vascular infiltration (p < 0.01). In patients who had strongly positive P-LAP staining, the disease-free interval was significantly lower than in patients who had negative or weakly positive P-LAP staining (p < 0.01). Multivariate analysis demonstrated that strongly immunoreactive P-LAP (odds ratio, 12.8; 95% confidence interval, 2.84–58.8; p < 0.01) and surgical stage (odds ratio, 8.78; 95% confidence interval, 2.77–27.8; p < 0.01) are independent prognostic factors. Conclusions: This study suggests P-LAP as an independent prognosticator of clinical outcome in patients with endometrial carcinoma. Therefore, assessment of the P-LAP status provides clinically useful prognostic information in patients with endometrial carcinoma.

Gonadotropin-producing pituitary adenomas are extremely rare in reproductive-age women. We report here a case of gonadotroph microadenoma with ovarian hyperstimulation. It was found in a 29-yr-old infertile Japanese woman with enlarged multicystic ovaries. The patient had an elevated basal serum estradiol level (up to 6755 pM, or 1840 pg/ml). Serum FSH and prolactin were mildly elevated (15.4 IU/liter, 1.4 nM or 31.4 ng/ml), whereas LH was low (0.5 IU/liter). The FSH level was paradoxically elevated in response to TRH administration. Dynamic magnetic resonance imaging revealed a pituitary microadenoma. Daily administration of bromocriptine, a dopamine agonist, normalized the ovarian size, and the patient ovulated naturally. She conceived after 3 months of bromocriptine therapy and delivered a normal child. She underwent elective transsphenoidal pituitary surgery, 3 yr after the delivery. Immunostaining of the resected tumor showed that 80% and less than 5% of the tumor cells stained for FSH-beta and prolactin, respectively. Furthermore, RT-PCR suggested that dopamine type 2 receptor was expressed in the adenoma. Gonadotroph microadenoma should be considered in women with spontaneous ovarian hyperstimulation, even if they have no neurological symptoms or marked pituitary enlargement. In such cases, bromocriptine therapy may be an alternative to pituitary surgery.

Adipocyte-derived leucine aminopeptidase (A-LAP) is a novel zinc-metallopeptidase involved in angiotensin II (AngII) metabolism, cell migration and antigen presentation. These functions are implicated in the progression of cancer, whereas A-LAP expression and involvement have not been studied in any type of cancer. We investigated the expression of A-LAP in endometrial cancer as well as its association with angiogenesis and clinicopathological features. Immunohistochemical staining of 58 endometrial endometrioid adenocarcinoma specimens revealed that 37 were A-LAP immunoreactive. We also found that A-LAP staining correlated with histological tumor grade in a significant and reverse manner. In addition, serum CA-125 levels in patients with A-LAP positive cancers were significantly higher. However, contrary to our hypothesis that A-LAP suppresses angiogenic activity via AngII metabolism, A-LAP expression was not associated with the microvessel count determined by CD34 immunostaining. Our results suggest that A-LAP is involved in endometrial cancer cell growth and differentiation. However, further studies, especially of the biological roles of A-LAP, are required to confirm this notion.

Dipeptidyl peptidase IV (DPPIV)/CD26 is a cell surface aminopeptidase. This study investigated the expression and localization of DPPIV in endometrial endometrioid adenocarcinomas of different grades.Immunohistochemical analysis was performed by using DPPIV and regulated on activation, normal T-cell expressed and secreted (RANTES) specific monoclonal antibodies. Cell proliferation was evaluated by bromodeoxyuridine (BrdU) uptake assay.Immunohistochemical analyses showed that DPPIV was strongly or moderately stained in glandular cells of the normal secretory phase. In endometrial adenocarcinoma, the DPPIV expression decreased with advancing grade (P.01). Furthermore, RANTES, one of the possible DPPIV substrates, was highly expressed in all grades of endometrial adenocarcinoma cells. The addition of RANTES to endometrial adenocarcinoma cells increased proliferation in a concentration-dependent manner.DPPIV is expressed in normal endometrial glandular cells, but its expression in endometrial adenocarcinoma is down-regulated with increasing grade. Our data also suggest a regulatory role of this ectoenzyme in neoplastic transformation and progression of endometrial adenocarcinomas possibly by degrading certain bioactive peptides such as RANTES.

The object of this study is to evaluate the radiological and pathological changes in the sinus of an experimental arteriovenous fistula of the rat. Twenty-five male Sprague-Dawley rats, including two control rats, were used for this study. A venous hypertension model in the transverse sinus was induced by means of anastomosis of a common carotid artery (CCA) to the ipsilateral external jugular vein (EJV). Rats were sacrificed 11 to 42 weeks after the procedure, then histopathological and immunohistopathological examinations were performed for the resected transverse sinus. Follow-up angiography was performed two to three weeks after the anastomosis in every case, and five months later in two rats. Patency of the anastomosed portion was confirmed in 12 of the 23 anastomosed rats. An ipsilateral carotid angiogram demonstrated a high-flow arteriovenous (AV) shunt from the CCA to the sigmoid-to-transverse sinus and draining into the contralateral juglar vein. A contralateral angiogram displayed a steal phenomenon via the communicating artery. Histopathologically, the vein of the anastomosed portion and the transverse sinus were markedly dilated in with cases. There was a thickening the connecting tissue and a proliferation of fibroblast in four (50%) of the eight cases. Thrombus formation in the transverse sinus was found in one case. VEGF stained strongly in the endothelial hypertrophied area and in fibrous connective tissue around the transverse sinus compared to the control sinuses. Our results from this long-term observation of the radiological and pathological changes in the sinus exposed to hypertension resembled the clinical findings of a dural AV fistula.

In pancreatic cancers, K-ras mutations have been found frequently (80%-100%), and they could be a good marker to detect tumor DNA in the plasma. Several studies have indicated that polymerase chain reaction/restriction fragment length polymorphism (PCR/RFLP) analysis of K-ras mutation was a useful method for the detection of hepatic and lymph node metastasis of pancreatic cancer. However, this method sometimes exhibited false-positive results, and the rate of K-ras mutation might thus be overestimated in these tissues. To diagnose pancreatic cancer correctly at an early stage, we attempted to detect tumor DNA in the plasma of pancreatic cancer patients using a more sensitive and specific method.We examined 28 pancreatic cancer patients using a sensitive mutation-specific mismatch ligation assay for K-ras gene mutations in primary tumors and paired plasma samples.K-ras gene mutations were detected in 26 of the 28 (93%) pancreatic cancers. We also found the same mutations in 9 of these 26 (35%) patients in their plasma DNA. This mutation was found even in the plasma of patients with TNM stage II cancer.Genetic alterations present in the tumors of pancreatic cancer patients can be detected in their plasma, and this approach is potentially applicable for cancer screening and the monitoring of this deadly disease.