Moriatsu Takada

3D-CT diagnosis for ingested foreign bodies.

Ingested foreign bodies can be hard to diagnose but cannot be missed. We report two cases where helical computed tomography (three-dimensional computed tomography) was used for the effective preoperative diagnosis (swallowed fish bone-induced perforation of sigmoid colon and a case of ileus caused by ingested PTP [press-through package]). Other traditional diagnostic methods could not identify the foreign bodies. Three-dimensional computed tomography is useful for the diagnosis of foreign body ingestion and should be used for the difficult cases.

Suppression of human pancreatic carcinoma cell growth and invasion by epigallocatechin-3-gallate.

Introduction The consumption of green tea is associated with a lower risk of several types of human carcinomas. A number of studies have focused on the possible mechanisms of cancer prevention by tea extracts, especially polyphenols such as epigallocatechin-3-gallate (EGCG). Aims and Methodology Green tea–derived EGCG was tested in human pancreatic carcinoma cells. The cells (PANC-1, MIA PaCa-2, and BxPC-3) were treated with different doses of EGCG (0, 25, 50, 100, and 200 &mgr;mol/L) for 48 hours in culture medium. Proliferation of pancreatic carcinoma cells was measured by means of the WST-1 colorimetric assay. For the study of cell invasion, the cells were incubated with 100 &mgr;mol/L EGCG for 2 hours. Then, the cells were added into the cell insert, coated with Matrigel basement membrane matrix. After incubation at 37°C for 24 hours, the cells that had invaded through the Matrigel were counted visually under the microscope. Results The growth of all three pancreatic carcinoma cells was significantly suppressed by EGCG treatment in a dose-dependent manner. EGCG treatment caused significant suppression of the invasive ability of pancreatic carcinoma cells PANC-1, MIA PaCa-2, and BxPC-3 but did not affect the cell cycle protein cyclin D1. Conclusion EGCG may be a potent biologic inhibitor of human pancreatic carcinomas, reducing their proliferative and invasive activities.

Comparative study of electrothermal bipolar vessel sealer and ultrasonic coagulating shears in laparoscopic colectomy

BackgroundAs an alternative to suture ligatures of the vessels, an electrothermal bipolar vessel sealer (EBVS) was recently developed. Meanwhile, ultrasonic coagulating shears (UCS) have been widely used clinically to provide hemostatic cutting in laparoscopic procedures. We conducted a comparative study to investigate the relative advantages of these two instruments.MethodsThe study included 30 patients with colon cancer who underwent laparoscopic colectomy using either the EBVS or the UCS. We performed a comparative analysis of the instruments by viewing videotapes showing their application in laparoscopic transverse colectomy and sigmoidectomy.ResultsAverage patient age was 71.6 ± 1.4 years. Average hospital stay was 14.2 ± 1.0 days. There were no statistical differences between the groups in age and hospital stay. The incidence of rebleeding was significantly lower in the EBVS group than in the UCS group for both surgical procedures (0.3 vs 1.2 in transverse colectomy, 0.3 vs 2.0 in sigmoidectomy, respectively). In addition, the required time for mesocolon dissection was also significantly shorter when the EBVS was used in both laparoscopic transverse colectomy and sigmoidectomy (7.9 vs 18.4, 15.0 vs 27.6, respectively).ConclusionThe use of the EBVS will enable surgeons to reduce the total operating time for laparoscopic colectomy.

Diagnostic imaging of early gallbladder cancer: retrospective study of 53 cases.

Abstract. To diagnose early gallbladder carcinoma is difficult but essential to improve the survival of the patients with this cancer. Fifty-three early gallbladder cancers were macroscopically divided into protruding and flat types. The diagnostic devises [ultrasonography (US), computed tomography (CT), and drip infusion cholangiography (DIC)] were compared for their ability of early detection. The specimens were examined cytologically for diagnosis during operation and the p53 protein was investigated. Thirty-three cases were of the protruding type, eighteen of the flat type, and two unclassified. Carcinoma tended to be missed when gallstones were present. Preoperative diagnosis of the flat type was difficult. Tumor location did not always correlate with the preoperative diagnosis. Of the misdiagnosed cases of the protruding type, half were missed with US and CT and were not visualized clearly by DIC. Among the flat type cancers, only three had no abnormal findings by diagnostic imaging. Cytologic examination was effective, and p53 was expressed only in early carcinoma, not in adenoma or dysplasia. Even in the presence of gallstones or cholecystitis, any abnormal findings should make one suspicious of gallbladder cancer. Cytology and p53 expression may be useful for the intraoperative diagnosis, and a combination of diagnostic methods is important.

Concurrent occurrence of gastric adenocarcinoma and duodenal neuroendocrine cell carcinoma: a composite tumour or collision tumours ?

BACKGROUND Neuroendocrine cell (NEC) carcinoma is occasionally accompanied by adenocarcinoma but the relationship between these two morphologically distinct tumours is unclear. Two hypotheses have arisen regarding the mechanism for the association of adenocarcinoma and NEC carcinoma. One is that both are derived from a common multipotential epithelial stem cell. The second hypothesis is that adenocarcinoma and NEC carcinoma arise from a multipotential epithelial stem cell and a primitive NEC, respectively. AIMS To elucidate the relationship between the two histologically distinct tumours, adenocarcinoma of the stomach and NEC carcinoma of the duodenum. PATIENT/METHODS We present a case in which the tumour extended across the pyloric ring, the gastric portion of which revealed adenocarcinoma while the duodenal portion showed argyrophil NEC carcinoma. The two histologically distinct lesions of the tumour were examined by immunohistochemistry and genetic analysis of p53. RESULTS The gastric region was negative for chromogranin A staining but positive for carcinoembryonic antigen (CEA) staining. In contrast, the duodenal region was positive for chromogranin A but negative for CEA. All tumour regions showed a point mutation in p53 gene at exon 7 (GGC (glycine)→GTC (valine) at codon 245). The distal portion of the duodenal tumour showed an additional point mutation inp53 gene at exon 5 (GCC (alanine)→GTC (valine) at codon 129). CONCLUSIONS The two histologically distinct tumours, adenocarcinoma of the stomach and NEC carcinoma of the duodenum, appear to be derived from a common epithelial cell.

Activation of macrophage-associated molecules after brain death in islets.

Islet transplantation is now established as an optional treatment for type I diabetes. However, rates of insulin independence in islet transplant recipients are still low. Although the major source of allograft is derived from brain-dead patient, the nonphysiologic state of brain death (BD) deteriorates organs such as liver and kidney. To determine the effects of BD on islets, a rodent model of BD has been used. Histologically, islets of BD rats showed decreased permeability and impaired integrity of the cell membranes. Flow cytometric analysis showed that CD11b/c-positive cells within islets were slightly increased in BD. This result suggests that BD induces macrophage infiltration into the islets. Moreover, RT-PCR revealed significant augmentation of macrophages-associated inflammatory molecules (IL-1β, IL-6, TNF-α, and MCP-1) in islets from a BD donor. Inducible nitric oxide synthase (iNOS) was weakly expressed, although not reaching statistical significance compared with control. Our results indicate that islets from a BD donor are immunologically activated and have a potential risk factor for early graft loss and a poor long-term function of grafts in clinical setting of islet transplantation. Immunomodulation, to eliminate intraislet immunocytes and/or activated macrophage-associated molecules, might be necessary for the better outcome after islet graft from BD donors.

The significance of CD44 in human pancreatic cancer: I. High expression of CD44 in human pancreatic adenocarcinoma.

The CD44 cell surface glycoprotein, which is the adhesion molecule of lymphocytes, has been suggested to be an important factor for the metastatic potential and invasive ability of cancer. We demonstrated the expression of CD44 in human pancreatic adenocarcinoma cells and normal cells by using flow cytometry (FACS-can) and immunohistological staining. CD44 was highly expressed in human pancreatic adenocarcinoma cells, while it was little expressed in normal human pancreas cells. These results indicate that the quantitative analysis of CD44 expression may be a useful diagnostic tool for pancreatic cancer.

Inhibitory effect of epigallocatechin-3-gallate on growth and invasion in human biliary tract Carcinoma cells

Based on recent evidence that tea consumption contributes to a decreased incidence of human carcinomas, a number of investigators have focused on the mechanisms of cancer prevention by tea extracts, especially green tea polyphenols. Epigallocatechin-3-galIate (EGCG) is a representative polyphenol that inhibits the activity of the cyclin-dependent kinases of cdk2 and cdk4. This suggests that EGCG may exert its growth-inhibitory effects through modulation of G1 regulatory proteins such as cdk2 and cdk4. The human biliary tract carcinoma cells (TGBC-2, SK-ChA-1, and NOZC-1) were treated with different doses of EGCG (0, 25, 50, 100, and 200 µM) for 48 hours in cell medium. Cell proliferation was analyzed by WST-1 colorimetric assay. For the cell-invasion analysis, the cells were incubated with 100 µM of EGCG for 2 hours. The cells were then added into a Matrigel-coated Cell Insert. After incubation at 37°C for 24 hours, the cells visible through the Matrigel were counted under the microscope. All human biliary tract cancer cells studied showed a significant suppression of cell growth by EGCG treatment in a dose-dependent manner (27.2%, 16.0%, and 10.1%, in TGBC-2, SK-ChA-1, and NOZC-1, respectively, at the dose of 200 µM). EpigaIlocatechin-3-gallate treatment also produced a significant suppression of invasive ability of the carcinoma cells (12.6%, 11.2%, 7.9%, in TGBC-2, SK-ChA-1, and NOZC-1, respectively, at a dose of 100 µM). These data indicated that EGCG might be a potent biological inhibitor of human biliary tract cancers, reducing their proliferative and invasive activities.RésuméBasé sur des indices récents semblant indiquer que la consommation en thé contribue à une incidence moindre de cancers chez l’homme, un certain nombre de chercheurs se sont intéressés aux mécanismes de prévention du cancer par les extraits de thé, et notamment les polyphénoles du thé vert. L’épigallocatéchine-3-gaIlate (EGCG) est un polyphénole représentatif qui inhibe l’activité de cdk2 et de cdk4. Ceci suggère que PEGCG pourrait exercer ses effets inhibiteurs sur la croissance par la modulation des protéines de régulation Gl telles que le cdk2 et le cdk4. Des cellules carcinomateuses des voies biliaires humaines (TGBC-2, SK-ChA-1, NOZC-1) ont été traitées par des doses variables d’EGCG (0, 25, 50, 100 et 200 µM) pendant 48 heures dans un milieu cellulaire. Cette prolifération cellulaire a été analysée par un essai colorimétrique WST-1. Pour l’analyse de l’invasion cellulaire, les cellules ont été incubées pendant deux heures dans 100 µM d’EGCG. Ensuite, les cellules ont été ajoutées à un appareil d’insertion cellulaire enduit de Matrigel. Après incubation à 37°C pendant 24 heures, les cellules envahies à travers le Matrigel ont été comptées visuellement sous microscope. Toutes les cellules humaines étudiées provenant d’un cancer des voies biliaires ont montré une suppression significative de la croissance cellulaire par le traitement par l’EGCG d’une manière dose-dépendante (27.2%, 16.0% et 0.1%, pour, respectivement, TGBC-2, SK-ChA-1 et NOZC-1, aux doses de 200 µM). Le traitement par l’EGCG a également produit une suppression significative de la capacité d’invasion (12.6%, 11.2%, 7.9%, pour, respectivement, TGBC-2, SK-ChA-1 et NOZC-1 à la dose de 100 µM). Ces données indiquent que l’EGCG pourrait être un inhibiteur biologique puissant du cancer des voies biliaires humaines, en réduisant beaucoup l’activité proliférative et invasive de ce cancer.ResumenRecientemente se ha demostrado que el consumo de té disminuye la frecuencia del cáncer en el ser humano. De ahí, que numerosos investigadores, intentado explicar el mecanismo preventivo del té, se hayan dedicado al estudio de sus extractos, sobre todo por lo que a los polifenoles del té verde se refiere. El Epigallocatectin-3 gallato (EGGG) constituye el más representativo de todos ellos inhibiendo la actividad de los cdk2 y cdk4. Estos hechos sugieren que el EGGG inhibe el crecimiento merced a la modulación de las proteínas reguladoras de la fase Gl, tales como cdk2 y cdk4. Células neoplásicas del árbol biliar humano (TGBC-2, SK-ChA-1, NOZC-1) fueron tratadas con diferentes concentraciones de EGGG (0, 25, 50, 100 y 200 µM) en un cultivo celular apropiado, durante 48 horas. La proliferación celular se estudió mediante el método colorimétrico WST-1. Para analizar la invasión celular se incubaron las células con 100 µM de EGGG durante 2 horas. Después, las células se trasladaron a un “Matrigel coated Cell Insert” y tras incubación a 37°C, durante 24 horas, las células que invadían el gel matricial (Matrigel) se contaron visualmente mediante microscopía. Todas las células neoplásicas procedentes del árbol biliar humano mostraron una supresión significativa (dosis dependiente) de su crecimiento, al ser sometidas a un tratamiento (dosis de 200 µM) con EGGG. (Para las TGBC-2 representó el 27.%, para las SK-ChA-1 el 16% y el 10.1% para las NOZC-1). El EGGG también provocó una supresión significativa (a dosis de 100 /utM) por lo que a la capacidad invasiva de dichas células se refiere y que fue del 12.6% para las células TGBC-2, del 11.2% para las SK-ChA-1 y del 7.9% para las NOZC-1. Los hallazgos muestran que posiblemente, el EGGG sea un potente inhibidor biológico del cáncer de árbol biliar, reduciendo su actividad tanto proliferativa como invasiva.

Three-dimensional reconstruction of biliary tract using spiral computed tomography for laparoscopic cholecystectomy

To increase the safety of laparoscopic cholecystectomy, we have analyzed the biliary tract in a three-dimensional fashion. The union of the biliary ducts was studied in 50 patients using spiral computed tomography (CT) after intravenous infusion cholangiography. Depending on the union and course of the cystic duct to the common bile duct, cystic duct anatomy was classified into six categories: ipsilateral gallbladder side (type I) and contralateral side with anterior course (type II); contralateral side with posterior course (type III); intrahepatic side (type IV); intrapancreatic side (type V); and unclassified (type VI). The length of the cystic duct was also determined. The cystic duct was identified in 42 cases (84%); 30 cases (60%) were type I, 9 cases (18%) type III, and a single case (2%) of types II, IV and V, respectively. The length of the cystic duct was ≥2 cm in 30 cases that had a shorter operating time compared to 12 cases with a cystic duct <2 cm (p<0.01). In conclusion, three-dimensional reconstruction of the cystic duct anatomy using spiral CT provides simple classification of bile duct anatomy, and this preoperative information may increase the safety of laparoscopic cholecystectomy.RésuméPour augmenter la sécurité de la cholécystectomie laparoscopique, nous avons analysé, en trois dimensions, la jonction des voies biliaires chez 50 patients en utilisant la tomodensitométrie (TDM) spiralée après cholangiographie intraveineuse. Selon le type d’union et le trajet du canal cystique (CC) vers la voie biliaire principale, on a classé l’anatomie biliaire en cinq catégories: vésicule biliaire (VB) du même coté (type I), VB du coté opposé avec trajet antérieur du CC (type II), VB du coté opposé avec trajet postérieur du CC (type III), VB du coté intrahépatique (type IV), VB du coté intrapancréatique (type V) et VB hors catégorie (type VI). La longueur du CC a également été déterminée. Le CC a été identifié dans 42 cas (84%); respectivement, type I dans 30 cas (60%), type III dans 9 cas (18%), et un seul cas (2%) des types II, IV et V. La longueur du CC a été de 2 cm ou plus dans 30 cas, avec un temps d’opération plus court comparé aux 12 cas où le canal cystique était plus court, moins de 2 cm (p<0.01). En conclusion, la reconstruction en trois dimensions de l’anatomie du CC par TDM spiralée pourrait fournir une classification simple de l’anatomie biliaire. En addition, cette information en préopératoire pourrait augmenter la sûreté de la cholécystectomie laparoscopique.ResumenCon objeto de incrementar la seguridad de las colecistectomías laparoscópicas hemos analizado tridimensionalmente el árbol biliar. La disposición de los conductos biliares se estudió en 50 pacientes merced a una tomografía computarizada helicoidal tras inyección intravenosa colangiográfica. De acuerdo con la dirección y situación anatómica de la desembocadura del conducto cístico en el colédoco pueden diferenciarse 5 tipos: Tipo I, cuando el c. cístico se sitúa en el lado ipsilateral de la vesícula; Tipo II cuando se sitúa en el lado contralateral siguiendo una dirección anterior; Tipo III se sitúa en el lado contralateral pero discurre en un plano posterior; Tipo IV el c. cístico es intrahepático; Tipo V en situación intrapancreática y Tipo VI inclasificable. También evaluamos la longitud del c. cístico. Se consiguió identificar el cístico en 42 casos (84%); 30 (60%) correspondieron al tipo I, 9 (18%) al tipo III y uno solamente (2%) a cada uno de los tipos II, IV y V. La longitud del c. cístico fue igual o mayor a 2 cm en 30 casos lo que redundó en una reducción del tiempo operatorio, comparado con el empleado en los 12 casos de cístico corto<2 cm (p<0.01). En conclusión, la reconstrucción tridimensional de la anatomía del conducto cístico mediante tomografía computarizada helicoidal permite establecer una clasificación simple del árbol biliar, proporcionando una información preoperatoria que incrementa la seguridad de la colecistectomía laparóscopica.

Characterization of islet-infiltrating immunocytes after pancreas preservation by two-layer (UW/perfluorochemical) cold storage method.

Clinical islet transplantation offers the prospect of good blood glycemic control without major surgical risks. Nevertheless, long-term function of the transplanted islets is seldom appreciated because rejection is followed by the graft failure. Although it has been implicated that islets have high immunogenicity, characterization of the islet-infiltrating immunocytes, such as leukocytes and macrophages, has not been extensively studied. Rat islets were isolated by the collagenase digestion method and separated by handpicking under the microscope. The islets were further dispersed into individual cells for flow cytometric analysis. Monoclonal antibodies directed toward T cells, B cells, and macrophages as well as ICAM-1, and MHC class I and II were used to enumerate cells. Pancreatic islets contained 6.3 +/- 2.9% immunocytes; T cells (39.6 +/- 4.2%), B cells (44.7 +/- 5.8%), and macrophages (1.7 +/- 0.6%). MHC class I was expressed on 85.6 +/- 2.8%, MHC class II on 36.8 +/- 2.9%, and ICAM-1 on 39.9 +/- 7.0%. The results of islets from preserved pancreas also showed the same tendency. As these islet-infiltrating immunocytes within the grafts may contribute to the rejection, one potential strategy to prevent early graft loss might start to eliminate or inactivate the islet-infiltrating immunocytes.