大宮 直木

Host genetic susceptibility may influence gastric carcinogenesis caused by Helicobacter pylori infection. We aimed to clarify the relationship of interleukin (IL)-8 polymorphism with the risk of atrophic gastritis and gastric cancer. We examined IL-8 -251 T > A, IL-1B -511 C > T, and IL-1RN intron 2 polymorphisms in 252 healthy controls, 215 individuals with atrophic gastritis, and 396 patients with gastric cancer. We also investigated the effect of the IL-8 polymorphism on IL-8 production and histologic degree of gastritis in noncancerous gastric mucosa. Although no correlation was found in the analysis of the IL-1B and IL-1RN polymorphisms, IL-8 -251 A/A genotype held a higher risk of atrophic gastritis [odds ratio (OR), 2.35; 95% confidence interval (CI), 1.12-4.94] and gastric cancer (OR, 2.22; 95% CI, 1.08-4.56) compared with the T/T genotype. We also found that the A/A genotype increased the risk of upper-third location (OR, 3.66; 95% CI, 1.46-9.17), diffuse (OR, 2.79; 95% CI, 1.21-6.39), poorly differentiated (OR, 2.70; 95% CI, 1.14-6.38), lymph node (OR, 2.50; 95% CI, 1.01-6.20), and liver metastasis (OR, 5.63; 95% CI, 1.06-30.04), and p53-mutated (OR, 1.91; 95% CI, 1.13-3.26) subtypes of gastric cancer. The A/A and A/T genotypes were significantly associated with higher levels of IL-8 protein compared with the T/T genotype. Neutrophil infiltration score was significantly higher in the A/A genotype than in the T/T genotype. In conclusion, we showed that the IL-8 -251 T > A polymorphism is associated with higher expression of IL-8 protein, more severe neutrophil infiltration, and increased risk of atrophic gastritis and gastric cancer.

OBJECTIVE: The purpose of our investigation was to determine the usefulness of digital radiography (DR) for diagnosing the depth of invasion of esophageal carcinoma. METHODS: We evaluated 59 patients with esophageal carcinomas who underwent DR. During continuous DR in tangential views, the most distended image of the esophagus was chosen. Percent esophageal stenosis (PES) was based on the diameter across the lesion of maximal narrowing and the average of the normal oral and anal side diameters. The maximal thickness of the tumor was measured on sequentially prepared specimens. We evaluated whether the percent of esophageal stenosis correlated with the maximal thickness of the tumor on histologic findings. Receiver-operating characteristic (ROC) curves were constructed to establish the cut-off level for PES in diagnosing the depth of tumor invasion. Accuracies for the depth of the invasion were calculated based on PES using DR. For the accuracy rate, DR was compared with endoscopy and endoscopic ultrasonography (EUS). RESULTS: There was a close correlation between PES and pathological thickness of the tumor. PES values (mean+/-S.D.) were 2.45+/-0.75% in Tis and T1a tumors, 13.3+/-10.9% in T1b tumors, 35.2+/-11.1% in T2 tumors, 55.2+/-18.1% in T3 tumors, and 86.1+/-7.5% in T4 tumors. Using the ROC analysis, 12.5, 37.5, and 44.4% were the highest cut-off values of PES for differentiating < or =T1a, < or =T1b, and < or =T2 tumors. Regarding T staging, 45 (76%) of 59 lesions were staged correctly with EUS, whereas 47 (80%) were staged correctly with DR. CONCLUSION: DR is useful for diagnosing the depth of the invasion because esophageal stenosis calculated using DR is an objective index of tumor infiltration. The accuracy rate of the depth of invasion with DR was as good as that of EUS.

AIM: To estimate the detectability of anomalous connection in pancreatobiliary disease (ACPBD) cases, measuring gallbladder wall blood flow (GWBF). METHODS: In the retrospective study, we enrolled 42 subjects with gallbladder wall thickening. GWBF velocity was determined as an average value of the peak velocity of color signals on the gallbladder wall, three times in each case. Based on the findings on endoscopic ultrasonography (EUS) or endoscopic retrograde cholangiopancreatography (ERCP), the 42 subjects were divided into 11 cases with ACPBD and 31 cases without ACPBD. In the prospective study, the subjects were 92 cases with gallbladder wall thickening. Using the cut-off level of the flow velocity obtained in the retrospective study, the usefulness of measuring GWBF velocity in diagnosing ACPBD was evaluated. RESULTS: In the retrospective study, imaging of GWBF was obtained in 40 of the 42 subjects. The mean GWBF velocity of the ACPBD cases was 29.4+/-3.9 cm/s (mean+/-SD), which was significantly different (P<0.0001; 95% CI 5.48-13.2) from that of the without ACPBD cases (20.1+/-5.9 cm/s). Based on this result, we prepared a receiver operating characteristic curve, and the cut-off level appropriate for diagnosing ACPBD was estimated to be 25 cm/s. In the prospective study, GWBF was detected in 86 of the 92 subjects. Based on the EUS or ERCP findings, the 92 subjects were divided into 15 cases with ACPBD and 77 cases without ACPBD. When a cut-off level of 25 cm/s was employed, ACPBD could be diagnosed with a sensitivity of 87.0% (13/15) and a specificity of 87.3% (62/71). CONCLUSION: Measurement of GWBF velocity, which is less invasive and provides objective values, is very useful for diagnosing ACPBD prior to the development of malignant tumors in cases with gallbladder wall thickening.

We performed magnifying endoscopy for patients with suspected gastric diseases. Among these patients, 67 patients with early gastric cancer and 31 benign gastric diseases were enrolled in this study. The patients with early gastric cancer included 46 differentiated tubular adenocarcinoma (33 mucosal cancer, 13 submucosal cancer) and 21 non-differentiated tubular adenocarcinoma (12 mucosal cancer, 9 submucosal cancer). The benign gastric lesions included 23 gastric ulcer or gastric ulcer scars, three gastritis, and five gastric adenomas. Small regular patterns were observed 39% in differentiated adenocarcinoma, 5% in undifferentiated adenocarcinoma, and 19% in benign gastric diseases. Irregular patterns were observed 37%, 52%, and 6%. Lack Divisible structure was observed 18%, 90%, and 10%. Abnormal vessels were observed 26%, 81%, and 16%. Small regular patterns were observed significantly more frequently in differentiated adenocarcinoma than in undifferentiated adenocarcinoma (P &lt 0.001). Lack of visible structure and Irregular patterns were observed significantly more frequently in undifferentiated adenocarcinoma than in differentiated adenocarcinoma (P &lt 0.001). In order to spread this useful endoscopy widely easy recognition of abnormality, histological backbone, and further technical developments in hardware and software should be required.

BACKGROUND: The differentiation of benign from malignant intraductal papillary mucinous tumors (IPMT) is often difficult even by various examination methods. We evaluated the qualitative and quantitative diagnostic ability of contrast-enhanced transabdominal ultrasonography (CE-US), mainly in differentiating benign from malignant tumors in patients with IPMT. PATIENTS AND METHODS: There were 21 patients with IPMT who underwent CE-US and endoscopic ultrasonography (EUS). Surgery was performed in all 21 patients. Pathological findings were 4 with carcinoma and 17 with adenoma. CE-US was performed using a contrast agent (Levovist; Tanabe, Osaka, Japan) consisting of galactose microbubbles and a small (0.1%) admixture of palmitic acid, and the following items were evaluated by the following procedure. (1) Two reviewers with experienced sonographic and endosonographic ability evaluated CE-US images before and after contrast enhancement and classified the enhancement effects into three grades. In addition, the presence or absence of enhancement effects by CE-US was compared with that of mural nodules visualized by EUS. (2) In all 21 patients, changes in intensity after contrast enhancement were quantitatively measured using an HDI Lab. HDI Lab was provided by ATL (Philips; Bothell, WA) and these software tools rapidly quantify image characteristics within multiple ROI (regions of interest) and make comparisons between several areas or images. In both the early and late phases, the post-enhancement intensity, difference between pre- and post-enhancement intensity, and the percentage change ((post-enhancement value-pre-enhancement value)/pre-enhancement value) were compared between malignant and benign lesions, and the ability of CE-US to differentiate between benign and malignant lesions was evaluated in comparison with the ability of EUS to diagnose the degree of malignancy. RESULTS: (1) In both the early and the late phases, both reviewers observed enhancement effects in all 21 patients. And both reviewers observed mural nodules by EUS in all 21 patients. (2) In all 21 patients who underwent resection of IPMT, the intensity increased in both the early and late phases. When the patients with carcinoma were compared with those with adenoma, the post-enhancement intensity was significantly higher, and the difference between pre- and post-enhancement intensity and the percentage change in the early phase and the late phase was significantly more marked in the carcinoma group (p= 0.019, p= 0.002, p= 0.015, p= 0.012, and p= 0.039, respectively). CONCLUSIONS: CE-US was useful for qualitatively diagnosing tumor lesions in patients with IPMT. Moreover, quantitative changes in intensity can be a parameter for the differential diagnosis of benign and malignant tumors.

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.

OBJECTIVES: Some subjects infected by Helicobacter pylori have enlarged folds in the gastric body, the precise mechanism of which remains obscure. The aim of this study was to clarify the association of tumour necrosis factor-alpha (TNFA) gene polymorphism with susceptibility to hyper-rugosity. We also examined the association of TNFA polymorphism with gastric carcinoma. SUBJECTS AND METHODS: Four hundred and seventy-two subjects (male/female = 351/121, aged 26-81 years) without gastric carcinoma (control group), and 300 patients (male/female = 218/82, aged 32-91 years) with gastric carcinoma. Barium meal roentgenograms were performed in 396 subjects in the control group and fold width was measured at the greater curvature of the middle portion of the gastric body. Fasting plasma anti-H. pylori IgG titres, pepsinogens (PGs) I and II were analysed, and TNFA -857 promoter polymorphism was distinguished by the 5' nuclease polymerase chain reaction assay and polymerase chain reaction restriction fragment length polymorphism using HincII in both groups. RESULTS: Adjusted odds ratios of TNFA -857 T/T genotype and H. pylori seropositivity for hyper-rugosity (fold width = 6.0 mm) were 6.7 (95% confidence interval (CI) 1.5-28, P < 0.01) and 18.2 (95% CI 4.2-78, P < 0.0001), respectively. There were no significant differences in any genotype or allele frequencies between the control and total gastric carcinoma group. In a subgroup of gastric carcinoma patients who were negative for the PG assay, however, the odds ratio of the T allele was 1.4 (95% CI 1.0-2.0, P < 0.05). CONCLUSION: The TNFA -857 T/T genotype and H. pylori infection were strongly associated with rugal hyperplastic gastritis. The TNFA -857 T allele may promote gastric carcinoma without severe atrophy.

BACKGROUND AND AIM: It is common knowledge that endoscopic ultrasonography (EUS) can accurately diagnose pancreatic diseases. Echoendoscopes for EUS are roughly classified into two categories, the mechanical radial scanning echoendoscope (MR-ES) and the electronic linear array echoendoscope, both of which have their merits and demerits. In 2000, a newly designed echoendoscope, the electronic radial scanning echoendoscope (ER-ES), appeared. The aim of the present study was to compare B-mode image quality between the ER-ES and the MR-ES in pancreatic diseases. METHODS: Patients with pancreatic diseases (30 cystic diseases and 22 solid diseases) underwent EUS with both ER-ES and MR-ES. The B-mode images obtained using both echoendoscopes were graded using a scoring system and statistically analyzed. The assessed point for cystic lesions was the existence of mechanical-noise-like ring-like artifacts derived using multiple reflections ('ring-down'), grating robe and so on, and that for solid lesions was the scale of penetration. The authors compared maneuverability, endurance and endoscopic images between the two types of echoendoscopes. RESULTS: The ER-ES had a significantly higher score than the MR-ES (P < 0.05) in the analysis of both cystic and solid diseases. There was no apparent difference as to maneuverability, endurance and endoscopic images. CONCLUSION: Ultrasound images acquired by ER-ES appear better compared with those acquired by MR-ES.

BACKGROUND AND AIMS: Interferon (IFN)-gamma and tumor necrosis factor (TNF) are predominant cytokines produced in the gastric mucosa of patients with Helicobacter pylori-infected gastritis. Several studies reported that IFN-gamma and TNF induced the synergistic effect on many cell lines. We attempted to clarify the apoptotic activity and the synergistic effect of IFN-gamma and TNF on the gastric epithelial cell, and whether IFN-gamma relates to soluble TNF receptors (sTNF-R) release from the gastric epithelial cell. METHODS: On the gastric epithelial cell line MKN45, cytotoxic and apoptotic effects of IFN-gamma and TNF were examined. Next, sTNF-R released in response to IFN-gamma and the protective effect of sTNF-R against the cytotoxic activity of TNF and IFN-gamma were examined by blocking the release of sTNF-R with a serine protease inhibitor such as phenylmethylsulfonyl fluoride. RESULTS: Interferon-gamma significantly decreased cell viability, but TNF decreased it only slightly. Interferon-gamma and TNF did not make a synergistic effect on cell viability and apoptosis. Interferon-gamma and TNF induced sTNF-R release from gastric epithelial cells. Phenylmethylsulfonyl fluoride significantly inhibited shedding of sTNF-R and a synergistic effect of TNF and IFN-gamma on apoptosis was observed. CONCLUSION: These results suggest that sTNF-R released by IFN-gamma regulate the injury on the gastric epithelial cell line induced by TNF.

Of 987 cases of gastric adenocarcinoma seen at Nagoya University School of Medicine, we found 6 rare, extremely well-differentiated advanced gastric adenocarcinomas that could not be diagnosed as malignant tumors with only H&E staining, even with repeated biopsies under preoperative endoscopy. The aim of this study was to determine whether an immunohistochemical method using p53 and Ki-67 antibody would be helpfulfor preoperative pathologic diagnosis. The cancer control cases were 16 cases of ordinary well-differentiated advanced gastric adenocarcinoma, while the gastritis control cases were 22 cases of Helicobacter pylori-positive chronic gastritis. The p53 labeling index and the localization of Ki-67+ cells showed that the special adenocarcinomas in biopsy specimens were distinct from the surrounding normal mucosa and chronic gastritis, but not from the cancer control cases. These methods are useful markers for preoperative pathologic diagnosis of extremely well-differentiated gastric adenocarcinoma, which sometimes is confused with regenerative atypical glands before operation.

1. Liposome-mediated transfection is useful due to no DNA constraints, lower immunogenicity and easy preparation. However, it has the disadvantage of low transfection efficiency. We aimed to test whether lipofection efficiency could be enhanced in gastrointestinal cell lines by the growth-promoting effect of insulin. 2. To assess the effect of insulin on lipofection efficiency and the cell cycle, expression of green fluorescent protein (GFP) and DNA distribution in gastric (MKN1), colonic (HT29) and pancreatic (BxPC3) carcinoma cell lines was analysed using flow cytometry. 3. The percentage of positive cells with GFP was significantly higher in MKN1 cells in culture medium with 5 mg/mL insulin than without insulin, whereas the percentage was the same in HT29 and BxPC3 cells with insulin as without insulin. The percentage of S phase fraction MKN1 cells with insulin was greater than without insulin, whereas the percentage of S phase fractions of HT29 and BxPC3 cells was the same with or without insulin. Lipofection efficiency correlated with the percentage of S phase fraction. 4. Insulin has the potential to enhance efficiency of lipofection into a sensitive cell line by increasing cellular proliferation.

BACKGROUND: With the advent of immunohistochemical analysis, the term "gastrointestinal stromal tumor" (GIST) was proposed to designate the largest category of primary nonepithelial neoplasms. EUS-guided fine needle aspiration (EUS-FNA) is useful for diagnosis of GISTs. The aim of this study was to evaluate the phenotyping of GISTs and diagnosis of malignant GISTs by using EUS-FNA with immunohistochemical analysis. METHODS: A diagnosis of GIST was made in 23 patients by using EUS-FNA with immunohistochemical analysis. The accuracy of EUS-FNA diagnosis compared with the EUS imaging alone was analyzed. Additionally, immunophenotyping of specimens obtained by EUS-FNA and surgical resection specimens was compared. Factors that were diagnostic for malignant GISTs were also analyzed. RESULTS: The overall accuracy for the diagnosis of malignant GIST was 78% (18/23) by EUS imaging alone and 91% (21/23) by histopathologic evaluation (H&E staining) of specimens obtained by EUS-FNA. In 21 of 23 cases (91%) the immunohistochemical expressions of c-kit, CD34, muscle actin, and S-100 coincided for the FNA and surgical specimens. The presence of mitotic cells (p = 0.011) and the Ki-67 labeling index (p < 0.0001) with respect to the FNA specimens were significant predictive factors for malignant GIST. For the diagnosis of malignant GIST, the accuracy, sensitivity, and specificity of EUS-FNA with the addition of Ki-67 immunohistochemical staining were 100%. CONCLUSIONS: EUS-FNA with immunohistochemical analysis is useful in the preoperative diagnosis of GIST. It provides abundant information on immunohistochemical subtyping and on the capacity of the tumor for cellular progression.

Hemolymph nodes (HLs) are unique lymph nodes, in that their lymphatic sinuses contain numerous erythrocytes. In this study, we compared the internal structure and immunologic function of HLs with those of ordinary lymph nodes (OLs) and the spleen. Electron microscopy revealed erythrocytes passing through the walls of blood vessels in the intermediate sinus area (IMSA) of a HL between expanded endothelial cell junctions. However, no direct communication was found between lymphatic sinuses and blood vessels. Numerous carbon particles appeared in the IMSA of HLs on 5 days after intravenous carbon particle injection, while OLs lacked particle deposition. Immunohistochemical studies showed that lipopolysaccharide (LPS) reached the IMSA of HLs and extravasated into medullary cords 4 hours after intravenous LPS injection, resulting in the appearance of more IgM-stained lymphocytes in the IMSA of HLs than in that of OLs on day 5. The ability of organs to produce antibodies was determined by counting the number of plaque forming colonies after intravenous injection of sheep red blood cells (SRBC). The HLs antibody-producing ability was between that of OLs and the spleen. These results suggest that HLs possess functionally open blood vessels in the IMSA and their immunologic capability is between that of OLs and the spleen. These findings suggests that HLs are lymphoid organs that have characteristics between those of the OLs and the spleen, both ultrastructurally and functionally.