Abstract: AIMS: The aim of this study is to evaluate the influence of dividing the inferior mesenteric artery (IMA) and preserving the left colic artery (LCA) on rectosigmoid cancer surgery. PATIENTS AND METHODS: Colonic blood flow at the proximal site of the anastomosis was measured by laser Doppler flowmetry in 96 patients with cancer of the rectum and sigmoid colon while clamping IMA or LCA. Results were analyzed with patient characteristics and postoperative complications. RESULTS: Blood flow was significantly decreased by either IMA or LCA clamping, and its reduction rate was 38.5 +/- 1.8%, ranged from 0 to 82.8%, or 16.4 +/- 1.8%, ranged from 0 to 66.2%, respectively. For multivariate analyses, aging and male gender were predictive factors of high blood flow reduction by IMA clamping. The reduction rate was significantly correlated with aging in male patients, while no such correlation was observed in women. Aging correlation in men was more significant in ultralow anterior resection cases. Three elderly male patients received IMA high ligation among 19 patients who demonstrated more than 50% blood flow reduction by IMA clamping. Among these, two patients, those who underwent ultralow anterior resection, suffered severe anastomotic ischemia. CONCLUSIONS: Colonic blood flow at the proximal site of the anastomosis was significantly decreased by either IMA or LCA clamping. Patients with high reduction by IMA clamping need intraoperative efforts to prevent anastomotic ischemia, particularly in elderly male patients who undergo ultralow anterior resection.

Abstract: BACKGROUND: After anterior resection for rectal carcinoma patients often complain of defaecatory dysfunction. The aim of this study was to examine the number of interstitial cells of Cajal (ICCs) and ganglion cells in the distal colon partially denervated by mesenteric transection in a rat model. METHODS: Four groups of rats were operated on, a control group and three experimental groups, with increasing degree of mesenteric transection to denervate the distal colon. Animals were killed after 3 months, and the denervated colon was investigated using immunohistochemistry with Kit and protein gene product 9.5 antibodies to detect ICCs and ganglion cells respectively. Reverse transcriptase-polymerase chain reaction for c-kit and histomorphometry of muscular thickness were performed. RESULTS: The total number of ICCs was significantly lower in the three operated models than in the control, particularly in the histological layers of the myenteric plexus and submucosal border. There was no difference in the number of ganglion cells or changes in muscular thickness between the models. CONCLUSION: The number of ICCs in rat distal colon denervated by mesenteric transection is decreased at 3 months after surgery.

Abstract: BACKGROUND AND AIM: Postoperative ileus (POI) is a transient bowel dysmotility following abdominal surgery. The effects of adrenergic blocking agents and celiac ganglionectomy were studied in rats to investigate the possible involvement of the adrenergic pathway in whole gastrointestinal (GI) transit in the early and late phases of POI. METHODS: After laparotomy, the terminal ileum was manipulated for 10 min. (51)Cr was administered into the stomach or colon immediately after surgery. In another group, (51)Cr was administered 24 h after surgery. Three hours after (51)Cr was administered, the rats were killed, and GI and colonic transit was calculated as a geometric center (GC). RESULTS: Gastrointestinal transit was significantly delayed 3 h after intestinal manipulation, compared with GI transit in rats that had anesthesia only. Three hours after intestinal manipulation, guanethidine (5 mg/kg) and yohimbine (3 mg/kg) significantly improved delayed GI transit. Celiac ganglionectomy also significantly improved delayed GI transit. Twenty-seven hours after intestinal manipulation, guanethidine, yohimbine and celiac ganglionectomy improved delayed GI transit induced by intestinal manipulation. Colonic transit was delayed 3 h after intestinal manipulation, and delayed colonic transit was partially restored within 27 h of intestinal manipulation. Guanethidine, yohimbine and celiac ganglionectomy improved delayed colonic transit 3 h and 27 h after intestinal manipulation. CONCLUSIONS: Adrenoceptors activated by intestinal manipulation impair the motility of the entire GI tract in both the early and the late phase of POI.

Abstract: Rho family GTPases play a pivotal role in the regulation of numerous cellular functions associated with malignant transformation and metastasis. To evaluate the role of these GTPases in colorectal cancer, the protein expression levels and activities of these proteins in matched sets of tumor and non-tumor tissues of surgical specimens were analyzed. The relationship between the protein levels and activities in tumor tissues to the clinicopathological features was also assessed. The expression levels of RhoA, active RhoA, Rac1, and active Rac1 in tumor tissues were higher than in normal tissues. The amounts of active RhoA protein in primary tumors correlated with lymph nodes metastasis. No relationship was noted between the protein expression levels and other clinicopathological findings. These findings suggest that the Rho family small GTPases are related to malignant transformation and progression of colorectal cancer and the activation of RhoA is associated with the lymph node metastasis.

Abstract: BACKGROUND: Lymphatic mapping and sentinel lymph node (SN) biopsy has rapidly replaced axillary lymph node dissection for clinically node-negative breast cancers. Because of a short follow-up period when the procedure was new, there were few reports of the clinical recurrence rate in breast cancer patients treated with SN biopsy. The present study attempts to clarify the occurrence of distant failure after SN biopsy, especially in breast cancer patients with SN micrometastasis. METHODS: The subjects consisted of 375 cases with clinically node-negative breast cancer, who had undergone SN biopsies. Chemotherapy and/or hormonal therapy was recommended based on the pathological primary tumor characteristics. The patients with SN micrometastasis also received adjuvant therapy equal to node-positive patients. RESULTS: Examinations of lymph nodes indicated metastases in 73 cases. Among the invasive cancers, 54 cases had macrometastasis, 19 cases had micrometastasis and 241 cases had a tumor free SN. The median follow-up period ws 30 months (range 6 to 66 months). Distant relapse rates per person-years were 0.3% in the cases with tumor free SN and 3.3% among the macrometastatic cases. However, systemic disease was not observed in the cases with SN micrometastasis. CONCLUSIONS: These results may show that upstaging due to SN investigation increases the number of cases who should receive anti-cancer drugs, and consequently reduces the distant relapse rate. Further studies in a large number of cases as well as longer follow-up are needed to determine the prognostic significance of SN micrometastasis.

Abstract: BACKGROUND: Anterior resection for rectal disease is associated with extrinsic autonomic denervation of the neorectum, which may influence the myenteric plexus, and subsequently the motility/defecatory status after operation. METHODS: A rat model with denervated neorectum was constructed. Colonic contractile activity in vivo, the amount of generic neuron marker (PGP 9.5) and nitric oxide synthase (NOS) were measured periodically. The responses of the muscle strip in each period to electrical field stimulation were evaluated using various neurotransmitters. RESULTS: In rats with denervated neorectum, giant migrating contractions (GMCs) of the distal colon, the number of fecal lumps per day and their small size, significantly increased in the early phase postoperatively, although both recovered in the late-phase period. The contractile response of the muscle strip of the denervated colon to acetylcholine was reduced throughout the period; however, contraction of the denervated colon under the addition of NO inhibitor (l-NAME) was enhanced significantly in the late-phase period, and recovered to the control level by atropine. Neuronal NOS, but not PGP 9.5 concentration, in the myenteric plexus at the distal denervated colon, significantly increased in the late-phase period. None of the above items differed from the control at other colonic portions throughout the period. CONCLUSIONS: Extrinsic autonomic denervation causes abnormal hyper-motility in the neorectum, which may be associated with multiple evacuations in the early phase postoperatively. Increased acetylcholine and the subsequent increase of neuronal NOS in the myenteric plexus may be an adaptive mechanism to compensate for such abnormal colonic motility after extrinsic denervation.

Abstract: BACKGROUND AND AIMS: Aberrant crypt foci (ACF) are thought to be preneoplastic lesions and are assessed by magnifying chromoscopy with methylene blue staining. The aim of this study was to evaluate the predictive value of rectal ACF recognized by conventional chromoscopy for colonic advanced neoplasms. METHODS: Total colonoscopy, involving rectal chromoscopy using indigo carmine with standard colonoscopies, was performed on 386 patients. Patients who showed no ACF were classified as Grade 0, and those who had 1-4, 5-9, and 10+ ACF were classified as Grades 1, 2, or 3, respectively. The correlation between ACF grading and the prevalence of colonic advanced neoplasm, any adenoma>or=1 cm in size and/or with villous or tubulovillous morphology, and/or with high-grade dysplasia or invasive cancer, was assessed. RESULTS: Sixty-three patients were classified as ACF Grade 0, 119 as Grade 1, 116 as Grade 2, and 88 as Grade 3. Colonic advanced neoplasm was observed in 4 patients (6.3%) for Grade 0, 43 (36.1%) for Grade 1, 61 (52.6%) for Grade 2, and 57 (64.8%) for Grade 3. As the ACF grade increased, the chance of a patient having a colonic advanced neoplasm increased. For multivariate analyses, compared with patients with Grade 0, those with Grades 1, 2, or 3 had a greater risk of colonic advanced neoplasm (odds ratio [OR] 9.18, 95% CI 3.08-27.33, OR 20.44, 95% CI 6.81-61.42, and OR 32.94, 95% CI 10.49-103.41, respectively). CONCLUSIONS: Chromoscopic assessment of rectal ACF by conventional techniques is useful for predicting colonic advanced neoplasms.

Abstract: Metastatic inefficiency is a phenomenon by which a majority of tumor cells is lost in the blood stream during the metastatic process. We investigated the effects of dexamethasone (DEX), a synthesized glucocorticoid, on the serum susceptibility of a colon carcinoma cell line, HT-29, with respect to metastatic inefficiency. The susceptibility to serum cytotoxicity of these carcinoma cells is possibly an important factor in metastatic inefficiency. In this study, we used glucocorticoid because it modifies the function of the plasma membrane and has been shown to enhance the hematogenous metastasis of some tumor cells. Using HT-29 cells that had been treated with DEX in vitro, the following factors were evaluated: the metastasis of intrasplenic injected cells; in vitro and in vivo proliferation; motility; the production of matrix metalloproteinases (MMPs); and the expression of the membrane complement regulatory proteins CD46, CD55, and CD59. The number of viable cells in the liver after an intraportal injection of tumor cells was determined by the expression of human beta-globin mRNA that is aberrantly expressed in HT-29 cells. In addition, we investigated 100% serum-induced proliferation, susceptibility, and apoptosis. Treatment with DEX was found to accelerate liver metastasis; here, the number of metastatic colonies and the weight of the liver were both significantly increased in DEX-treated HT-29 (HT-29DEX) cells. In contrast, there was no difference in terms of cell motility; the production of MMPs; or the expression of CD46, CD55, or CD59 between the HT-29 and HT-29DEX cells. The HT-29DEX cells exhibited enhanced proliferation in the serum, as well as resistance to cytotoxicity when exposed to 100% serum. In addition, DEX slightly inhibited serum-induced apoptosis. Finally, the expression of colon cancer-derived beta-globin mRNA was detectable 24 h after intravenous injection, but only in samples obtained from the HT-29DEX-, but not in those from the HT-29-inoculated mice. These results indicate that DEX reduced the metastatic inefficiency of the HT-29 cells, resulting in a hematogenous metastasis-prone phenotype. It is thus expected that the acquisition of resistance against serum cytotoxicity is among the mechanisms that contribute to the efficiency of hematogeneous metastasis.

Abstract: Slug, a member of the Snail family of transcription factors, plays a crucial role in the regulation of epithelial-mesenchymal transition (EMT) by suppressing several epithelial markers and adhesion molecules including E-cadherin. Recently, several studies have reported Slug to be expressed in breast carcinoma, oesophageal carcinoma accompanied with shorter survival. In this study, we first investigated expression of Slug mRNA in five colorectal carcinoma cell lines by reverse transcription-polymerase chain reaction. Furthermore, we investigated Slug and E-cadherin expression by immunohistochemistry in 138 patients with colorectal carcinoma. Slug mRNA was clearly expressed in four out of five colorectal carcinoma cell lines. Positive expression of Slug and E-cadherin was observed in 37 and 58% of cases, respectively. The positive expression of Slug was significantly associated with Dukes stage and distant metastasis (P = 0.0027 and 0.0007), and the positive expression of Slug had a significant impact on patient overall survival (P < 0.0001, log-rank test). Moreover, patients with positive expression of Slug and reduced expression of E-cadherin showed the worst prognosis (P < 0.0001, log-rank test). Multivariate analysis indicated that Slug expression was an independent prognostic factor. These results suggest that positive Slug expression in colorectal carcinoma patients may become a significant parameter of poor prognosis.

Abstract: PURPOSE: This study was designed to evaluate the feasibility and oncologic and functional outcomes of intersphincteric resection for very low rectal cancer. METHODS: A feasibility study was performed using 213 specimens from abdominoperineal resections of rectal cancer. Oncologic and functional outcomes were investigated in 228 patients with rectal cancer located <5 cm from the anal verge who underwent intersphincteric resection at seven institutions in Japan between 1995 and 2004. RESULTS: Curative operations were accomplished by intersphincteric resection in 86 percent of patients who underwent abdominoperineal resection. Complete microscopic curative surgery was achieved by intersphincteric resection in 225 of 228 patients. Morbidity was 24 percent, and mortality was 0.4 percent. During the median observation time of 41 months, rate of local recurrence was 5.8 percent at three years, and five-year overall and disease-free survival rates were 91.9 percent and 83.2 percent, respectively. In 181 patients who received stoma closure, 68 percent displayed good continence, and only 7 percent showed worsened continence at 24 months after stoma closure. Patients with total intersphincteric resection displayed significantly worse continence than patients with partial or subtotal resection. CONCLUSIONS: Curability with intersphincteric resection was verified histologically, and acceptable oncologic and functional outcomes were obtained by using these procedures in patients with very low rectal cancer. However, information on potential functional adverse effects after intersphincteric resection should be provided to patients preoperatively.

Abstract: Gastrointestinal neuroendocrine carcinomas (NECs) are extremely aggressive and poorly prognostic. We showed previously that human achaete-scute homologue gene 1 (hASH1), a basic helix-loop-helix transcription factor regulated by Notch, was aberrantly expressed in NECs. To date, no effective therapeutic strategies for NECs have been investigated. Notch, Wnt and Hedgehog (Hh) signalings are important for stem cell self-renewal and carcinogenesis in the gastrointestinal epithelium. In this study, we showed that Hh signaling was clearly upregulated in NECs in Gli1-dependent manner. Specific therapeutic effects of cyclopamine on NECs were also demonstrated. RT-PCR showed that among eight frozen samples (three NECs, one carcinoid tumor, three adenocarcinomas and one normal mucosa), the band intensities of Gli1 were the strongest in NECs, moderately strong in a carcinoid tumor, very weak in adenocarcinomas and undetectable in a normal mucosa. In real-time RT-PCR, the expression levels of Gli1 in NECs were 108.4, 28.6 and 16.3 times higher than that in an adenocarcinoma. In immunohistochemistry using 25 paraffin-embedded tissues, all twelve NECs and three of six carcinoid tumors showed positive stainings for Gli1, whereas six of seven adenocarcinomas were negative. In vitro, RT-PCR showed that NEC cell lines expressed Gli1 mRNA significantly. Administration of cyclopamine suppressed cell proliferation and invasion, and induced apoptosis in NECs. In cyclopamine-treated NECs, downregulation of Gli1, Ptch1, Snail and hASH1, and upregulation of E-cadherin were demonstrated at mRNA levels. Such effects were not observed in a Gli1-negative colonic adenocarcinoma cell line or in control alkaloid-treated NECs. Hh signaling may play a crucial role in the pathophysiology of NECs. Blockade of Hh pathway using cyclopamine or its synthetic derivatives might open an effective therapeutic strategy to NECs, not only by suppressing tumor viability but also by altering tumor cell nature.

Abstract: BACKGROUND/AIMS: Patients who have undergone anterior resection for rectal carcinoma often complain of anorectal and defecatory dysfunction postoperatively. The aim of this study was to examine the expression of interstitial cells of Cajal (ICCs) in the sigmoid colon used for constructing the neorectum after anterior resection of the rectum. METHODOLOGY: As the neorectum group, we assessed 12 patients with local and anastomotic recurrence or new neoplasm in the neorectum after anterior resection of the rectum. The control group consisted of 16 patients who underwent sigmoid colon resection for sigmoid colon carcinoma. All resected specimens were investigated with immunohistochemical staining, using c-kit antibody for ICCs. The correlation between the number of ICCs and defecatory symptoms was assessed for the neorectum. RESULTS: The total number of ICCs significantly decreased in the neorectum group as compared to the control group. In particular, a significant difference was noted between the two groups as to the number of ICCs found between the layers of the myenteric plexus in histological studies, as well as in the circular and longitudinal muscles. There was no correlation between the number of ICCs and the time interval from the initial anterior resection to the resection of the neorectum, nor was there any relationship between the number of ICCs and defecatory symptoms. CONCLUSIONS: The expression of ICCs in the neorectum was reduced in the early stages after anterior resection of the rectum. Expression of ICCs in the neorectum did not recover to preoperative levels over time.

Abstract: BACKGROUND: Primary chemotherapy for breast cancer is effective as postoperative adjuvant therapy. However, one of the critical disadvantages was a treatment delay for patients with progressive disease. The present study attempts to clarify quantitative parameters on MRI which can be used to predict the sensitivity to treatment in breast cancer patients. METHODS: The subjects consisted of 26 patients with invasive ductal breast cancer who received primary chemotherapy before surgery. The mean maximum tumor dimension was 3.3cm, and 21 cases had nodal involvements. Three cases demonstrated histological grade 3. Dynamic enhanced MRI was evaluated at three different time periods; prior to, in the midst of preoperative chemotherapy, and just before the initial operation. The signal intensity ratio (SIR) and early contrast uptake (ECU) were calculated, as well as the correlation between these dynamic data and the tumor reduction rates were analyzed retrospectively. P-values of less than 0.05 were considered to indicate statistically significant. RESULTS: Responders to chemotherapy had the significantly higher SIR and ECU values than non-responders (p=0.0454 and 0.0334, respectively). ECU value significantly decreased as tumor reduction by chemotherapy (p=0.0028). Pathological tumor dimension was significantly correlated with the tumor size estimated on presurgical MRI (p<0.0001). CONCLUSIONS: Our current series demonstrated the significant correlation between pretreatment MRI data and tumor reduction by chemotherapy in breast cancer patients. With these results, it seems possible to define good and non-responders prior to treatment.

Abstract: Thymidylate synthase (TS), dihydropyrimidine dehydrogenase (DPD), and vascular endothelial growth factor (VEGF) are associated with the effect of 5-fluorouracil (5-FU) based adjuvant chemotherapy. However, very few studies have investigated the relationship between these factors and 5-FU neoadjuvant chemotherapy for primary gastric cancer patients. In this study, we studied the correlation between these markers and the histological chemotherapeutic effect in advanced gastric cancer with neoadjuvant chemotherapy. METHODS: Sixty-two primary advanced gastric cancer patients were recruited into the study. One cycle of continuous infusion of 5-FU (300 mg/m2/day, 14 days) plus drip infusion of cisplatin (15 mg/m2/day, Day one and Day two) was performed as neoadjuvant chemotherapy. Histological chemotherapeutic responses of the resected specimens were classified into responders and nonresponders. TS, DPD, VEGF expressions both before and after neoadjuvant chemotherapy were examined immunohistochemically. RESULTS: There was an association between the TS-low group and the responders (p < 0.05); the DPD-low group and the responders in both biopsy and surgical specimens (p < 0.01). A combination of the low-TS and low-DPD group was further associated with responders (p < 0.01). The immunoexpressions of biopsied and surgical specimens were significantly associated with each other. CONCLUSION: Neoadjuvant chemotherapy for primary gastric cancer with one cycle of 5-FU and cisplatin was associated with histological findings in patients with low baseline TS and DPD. This dual determination may predict for efficacy of neoadjuvant treatment with these drugs.

Abstract: To decrease local recurrence and increase disease free survival, various preoperative therapies for patients with advanced rectal cancer have been studied. Cis-diamminedichloroplatinum (II) (CDDP) has become one of the most widely used cancer chemotherapeutic drugs. It has also been found to have radiosensitizing properties. In this experimental study, the efficacy of chemoradiotherapy using a novel CDDP suppository, and one with mixed micelles, was examined in a rabbit VX2 rectal tumor model. Rabbits were divided into four groups: control group, irradiation (R) group, CDDP suppository plus irradiation (CR) group, and mixed micelles plus CDDP suppository plus irradiation (CMR) group. Tumor growth ratios were reduced significantly in the CR and CMR groups as compared with the ratio in the control group. Microscopically, response rates of main tumors were 0%, 33.3%, 70.0%, and 91. 7%, respectively. The number of metastatic lymph nodes in the CR and CMR groups decreased significantly compared to the control group and the R group. The microscopic response rates of metastatic lymph nodes were 0%, 11.1%, 40.0%, and 41.7%, respectively. Lung metastases were observed in three rabbits in the R group, and in one rabbit in the CMR group. Tissue platinum concentrations both in tumors and in regional lymph nodes increased significantly when mixed micelles were used. Chemoradiotherapy using the CDDP suppository and mixed micelles was effective for local control in the rabbit VX2 rectal tumor model.

Abstract: PURPOSE: Gastrointestinal neuroendocrine carcinoma (NEC) is extremely aggressive, but its pathophysiologic features remain poorly understood. There have been no biologically specific markers for this disease. In this study, distinctive up-regulation of human achaete-scute homologue 1 (hASH1) in gastrointestinal NECs was clarified. EXPERIMENTAL DESIGN: Expression of hASH1 in NECs (n=10), carcinoid tumors (n = 10), other tumors (10 adenocarcinomas, 2 squamous cell carcinomas and 1 malignant lymphoma), and the corresponding normal mucosa were investigated by in situ hybridization, reverse transcription-PCR (RT-PCR), real-time RT-PCR, and immunohistochemistry. RESULTS: By in situ hybridization, mild to intense signals of hASH1 mRNA were detected in 9 of 10 NECs, but not in other tumors or normal mucosa, except for focally weak signals in one carcinoid tumor. RT-PCR showed strong expression of hASH1 in a small cell NEC, followed by a moderately differentiated NEC, and a carcinoid tumor, whereas it is undetectable in adenocarcinomas or normal mucosa. By real-time RT-PCR, the amounts of hASH1 mRNA in a small cell NEC were 16,600 times higher than those in adenocarcinomas and 110 times higher than those in a carcinoid tumor. Immunohistochemically, mammalian homologue of hASH1 was positive in 7 of 10 NECs but was negative in the other tumors. Pan-endocrine markers chromogranin A and synaptophysin were positive in almost all carcinoid tumors, in 4 and 7 of the 10 NECs, respectively. CONCLUSIONS: These findings revealed that hASH1 is distinctly up-regulated in gastrointestinal NECs. hASH1 may be used as a more sensitive and specific marker than conventional pan-endocrine markers for clinical diagnosis of gastrointestinal NECs.

Abstract: PURPOSE: The aim of this study was to determine whether denervation of the sigmoid colon during low anterior resection contributes to the postoperative motility characteristics of the neorectum and to the defecatory function of patients. METHODS: Sixty-seven patients who underwent either low or ultralow anterior resection for rectal cancer were evaluated. In accordance with the length of denervated neorectum, each patient was assigned to either the short-denervation or long-denervation group, determined by whether the inferior mesenteric artery was divided. Colonic propagated contraction was then measured by means of intraluminal pressure monitoring. Transit time was calculated with orally administered radiopaque markers. RESULTS: Propagated contraction down to the neorectum was significantly less common in the long-denervation group (14/36) than in the short group (12/15, P < 0.05), whereas spastic minor contraction at the neorectum was significantly more common in the long-denervation group (21/36) than the in short group (3/15, P < 0.05). Colonic transit time below the sigmoid colon was significantly longer in long group (6.4 hours) than in the short group (3.4 hours, P < 0.01). Although motility disorder of the neorectum was correlated with clinical defecatory malfunctions, including multiple evacuations, urgency, and soiling, no significant correlation was noted between the length of the denervated neorectum and the defecatory disorders. CONCLUSIONS: Motility of the neorectum following low anterior resection appears degraded by intraoperative maneuvers that cause denervation of the remnant sigmoid colon. Motility disorder of the neorectum, but not the length of the denervated neorectum causing the disorder, correlates well with several defecatory malfunctions. This finding suggests that postoperative defecatory disorder as a result of low anterior resection is caused by many factors in addition to denervation of the neorectum.

Abstract: Postoperative ileus (POI) is a transient bowel dysmotility that occurs following abdominal surgery. Several mechanisms have been proposed such as neural reflex and inflammatory changes. We focused on gastric motility after abdominal surgery in rats. To investigate the time course of gastric motility after surgery, gastric motility was continuously recorded before, during and after surgery. After laparotomy, terminal ileum was manipulated for 10 min. Gastric motility was recorded by a strain gauge transducer implanted on the serosal surface of the stomach. To investigate whether peripheral sympathetic nerve is involved in the pathogenesis of POI, effects of guanethidine and celiac ganglionectomy were tested on the postoperative gastric motility. Although isoflurane anaesthesia reduced the gastric motility to 40%, the motility recovered immediately when isoflurane was withdrawn. Intestinal manipulation reduced the postoperative gastric motility for 3-24 h after surgery, compared with preoperative levels. Guanethidine administration and celiac ganglionectomy restored the impaired gastric motility. Feeding increased the gastric motility in each group. It is suggested that the pathogenesis of postoperative gastric ileus induced by intestinal manipulation involves viscero-sympathetic pathways. Intestinal manipulation causes impaired gastric motility via inhibitory sympathetic efferent pathway. Feeding may improve the postoperative gastric motility.

Abstract: BACKGROUND: We investigated the correlation between operative procedures for rectal carcinoma and postoperative rectovaginal fistulas (RVF), and treatment for RVF. METHODS: The medical records of 161 female patients with rectal carcinoma were examined retrospectively with respect to the cause, incidence, and methods of treatment for RVF occurring after rectal cancer operations, and to the outcomes of gluteal-fold flap repairs for RVF. RESULTS: Of the 161 patients, 16 developed RVF clinically. The incidence of RVF was significantly higher in patients who were anastomosed by the double stapling technique (DST) and had concomitant resection of the vaginal wall. No statistical difference was found between the established diverting ostomy group and the no-stoma group. Six patients recovered by the establishment of a diverting ostomy only. The gluteal-fold flap technique was performed for 5 patients. No RVF recurrences were noted in these 5 patients. CONCLUSIONS: The incidence of RVF was higher in the patients who were anastomosed by DST or had concomitant resection of the vaginal wall. Although some RVFs heal with only fecal diversion, for patients in whom RVF is caused by involvement of the vaginal wall in the circular staple or intersphincteric resection, good results are obtained with the gluteal-fold flap repair technique.

Abstract: BACKGROUND: Breast microcalcifications are difficult to depict by ultrasound (US). However, recent advances in US equipment and the refinement of breast imaging techniques have improved the detection and characterization of small breast lesions. The present study attempts to determine whether US examination is able to demonstrate nonpalpable breast lesions associated with mammographically detected microcalcifications without mass density or distortion, and to evaluate the clinical reliability of US-guided procedures, especially in cases of ductal carcinoma in situ(DCIS)of the breast. METHODS: The subjects consisted of 73 patients with breast cancer diagnosed preoperatively as DCIS by stereotactic core needle biopsies, all of whom had microcalcifications without other abnormalities on mammography. The radiological appearance and size of the clustered microcalcifications were evaluated. US examinations were performed preoperatively, and the detection rates were assessed. Sonographically detected lesions underwent US-guided wire localization followed by surgical excision. RESULTS: The lesions associated with microcalcifications were identified sonographically in 54 of 73 cases (74%), and the pathological examination revealed breast cancer in all of the corresponding specimens. Lesions with linear-branching shape, segmental-linear distribution and category-5 calcifications on mammography had a high level of visibility on US. The US visible cases had a larger size of calcified area on mammography when compared with US invisible cases. Pathologically, the lesions were more frequently seen on US in cases with minimally invasive cancer or with comedo type DCIS. CONCLUSIONS: US examination is an effective method for identifying and localizing breast microcalcifications, and can be used as an alternative to stereotactic localization in selected patients with early breast cancer.

Abstract: To clarify the correlation between the expression level of thymidine phosphorylase (TP) and efficacy of doxifluridine (5'-DFUR) and 5-fluorouracil (5-FU), samples from 177 colorectal cancer patients who underwent curative resection were evaluated by immunohistochemical staining using a newly developed monoclonal antibody 1C6-203. Patients were randomly given either oral 5'-DFUR or 5-FU as postoperative adjuvant chemotherapy. In Dukes' C staged colon cancer patients treated with 5'-DFUR, better survival was observed in the high TP patients than the low TP patients (P=0.025 by the log-rank test). The observed 5-year survival rates were 91.2 and 74.8%, respectively. No correlation between TP expression and patient prognosis was detected in the 5-FU group. In Dukes' C stage colon patients with high TP expression, the 5'-DFUR group had slightly better survival than the 5-FU group. These findings suggest that TP may be a chemosensitive marker for 5'-DFUR as postoperative adjuvant chemotherapy for advanced colon cancer patients.

Abstract: A novel monoclonal antibody, SM/C-2.6, specific for mouse muscle satellite cells was established. SM/C-2.6 detects mononucleated cells beneath the basal lamina of skeletal muscle, and the cells co-express M-cadherin. Single fiber analyses revealed that M-cadherin+ mononucleated cells attaching to muscle fibers are stained with SM/C-2.6. SM/C-2.6+ cells, which were freshly purified by FACS from mouse skeletal muscle, became MyoD+ in vitro in proliferating medium, and the cells differentiated into desmin+ and nuclear-MyoD+ myofibers in vitro when placed under differentiation conditions. When the sorted cells were injected into mdx mouse muscles, donor cells differentiated into muscle fibers. Flow cytometric analyses of SM/C-2.6+ cells showed that the quiescent satellite cells were c-kit-, Sca-1-, CD34+, and CD45-. More, SM/C-2.6+ cells were barely included in the side population but in the main population of cells in Hoechst dye efflux assay. These results suggest that SM/C-2.6 identifies and enriches quiescent satellite cells from adult mouse muscle, and that the antibody will be useful as a powerful tool for the characterization of cellular and molecular mechanisms of satellite cell activation and proliferation.

Abstract: BACKGROUND AND AIMS: This study examined rectal cancers with lateral lymph node (LN) metastases and whether lateral lymph node dissection (LLD) with or without preoperative chemo-radiotherapy (XRT) benefits patients with rectal cancer. PATIENTS AND METHODS: A total of 452 consecutive cases of curatively resected pT2, pT3, and pT4 middle to lower rectal cancers were retrospectively analyzed. Of these, 265 patients underwent curative LLD and 155 XRT. Data were evaluated with respect to the cumulative percentage of survival. RESULTS: Lateral LN metastases were identified in 7.7% of patients. Of the pT3/pT4 extraperitoneal cancer patients 13.5/18.8% had lateral LN metastases. In the treatment of middle rectal cancers and pT2 extraperitoneal cancers LLD either with or without XRT did not improve survival rate. For the treatment of pT3/pT4 extraperitoneal tumors prior to the introduction of total mesorectal excision (TME) in 1994 LLD plus XRT yielded significantly better survival and local control than conventional surgery without LLD or XRT, although LLD alone did not improve either survival or local recurrence rates. Since 1995 TME with or without subsequent LLD has yielded favorable results for the treatment of extraperitoneal tumors. CONCLUSION: For the treatment of middle rectal cancers and pT2 extraperitoneal cancers LLD either with or without XRT does not improve survival rate. For pT3/pT4 extraperitoneal tumors, which are associated with a high incidence of lateral node metastasis, combining treatment modalities such as TME followed by LLD or XRT followed by TME may be considered.

Abstract: BACKGROUND: The correlation between postoperative defecatory status after anterior resection for rectal cancer and physiologic neorectal motility has not been well delineated. METHODS: Sixty patients who underwent anterior resection were examined. Motility of the neorectum was examined with 4-sensor intraluminal pressure monitoring, and segmental colonic transit time was determined with radiopaque Sitzmarks (Konsyl, Fort Worth, Texas) capsules. RESULTS: Twenty-eight patients experienced loss of propagated contraction waves down to the neorectum, which was closely correlated with a prolonged transit time through the neosigmoid colon and neorectum. In 26 patients, minor spastic waves were observed at the neorectum, which did not correlate well with the loss of propagated waves. The loss of propagation and the existence of spastic waves were significantly correlated with urgency of defecation and multiple evacuations. The latter was also associated with major soiling and with patients' assessments of impaired defecatory function. CONCLUSIONS: The physiologic motility of the neorectum is one of the factors responsible for postoperative defecatory function after anterior resection for rectal cancer.

Abstract: We report herein the case of a 70-year-old man who was found to have a gastrointestinal stromal tumor (GIST) in the stomach following sigmoid colon resection. Preoperative gastroscopic and barium examinations revealed a submucosal tumor, measuring 10 cm, on the upper part of the stomach. Using computed tomography (CT) images (i.e., computed tomographic volumetry) the doubling time of this tumor was calculated, accurately, as 3.3 months, which suggested a high growth rate and malignancy. A laparotomy and partial gastric resection were performed. Histologically, the tumor consisted of spindle-shaped cells with oval nuclei. In immunohistochemical studies, the tumor cells were positive with respect to c-kit, CD34, and vimentin, but negative with respect to smooth muscle actin and S-100 protein. There were 15-16 mitoses per 50 high-power fields (HPFs), and the Ki-67 antigen (MIB-1) index was 25.5% in the most active areas, which also indicated malignancy. The final pathological diagnosis of this tumor was malignant GIST. The patient was found to have hepatic metastasis 27 months after the surgery, and he subsequently received a hepatic subsegmentectomy. To our knowledge, there are very few reports concerning the growth rate of GISTs. Computed tomographic volumetry is useful for the follow-up of small or irregularly shaped gastric submucosal tumors, and for making decisions regarding surgical intervention.

Abstract: Centrally applied opioids delay gastric emptying and inhibit intestinal transit. However, the mechanism of inhibitory effects of central opioids on gastric motility still remains unclear. It also remains unclear which opioid receptor (mu, delta, and kappa) stimulation affects gastric motility. We studied the central effect of opioids on antral motility in conscious rats. A strain gauge transducer was implanted on the gastric antrum to record the circular muscle contractions. The area under the curve of the antral motility, calculated as a motility index, was evaluated before and after the intracerebroventricular (icv) injection of various opioid agonists in each rat. [D-Ala2, N-Me-Phe4, Gly5-ol] enkephalin (DAMGO, 0.1-10 nmol), a mu-opioid selective agonist, significantly inhibited antral motility in a dose-dependent manner (n=4). The motility index was significantly decreased to 47.3+/-10.8% (n=4) of controls at 20 min after icv injection of DAMGO (1.0 nmol). In contrast, [D-pen2, L-Pen5] enkephalin (DADLE, 1.0 nmol), a delta-opioid selective agonist, and U50,488 (1.0 nmol), a kappa-opioid selective agonist, had no significant effects on antral motility. Pretreatment with subcutaneous guanethidine (5 mg/kg) and propranolol (1 mg/kg), but not phentolamine (1 mg/kg), significantly antagonized the inhibitory effect of DAMGO (1.0 nmol). Reduced motility index induced by DAMGO (1.0 nmol) was restored from 48.7+/-3.5% to 88.6+/-10.9% (n=5) and 80.4+/-2.2% (n=5) by guanethidine and propranolol, respectively. Our findings suggest that central mu-opioid receptor has major inhibitory effects on antral motility in conscious rats. The inhibitory effects of mu-opioid receptors are mediated via sympathetic pathways and beta-adrenoceptors.

Abstract: BACKGROUND: The clinical practice of sentinel lymph node biopsy for breast cancer patients started in 1999 in our hospital, to obviate unnecessary axillary lymph node dissection. The present study examines the pathological false-negative cases on intraoperative sentinel lymph node investigations and evaluates their outcomes. METHODS: The subjects consisted of 183 cases with clinically node-negative breast cancer who had undergone sentinel node biopsy. When the sentinel node was noted to contain malignant cells intraoperatively, a complete axillary lymph node dissection was performed subsequently. The patients with tumor free sentinel nodes underwent no further axillary surgery. The pathological false-negative cases in this series were defined as patients with lymph node involvement which was revealed postoperatively, despite negative intraoperative sentinel node examinations. After these surgeries and/or adjuvant therapies, interval clinical evaluations were performed for all patients. RESULTS: Intraoperative diagnosis of the sentinel node was 96.2% accurate compared with the results of permanent sections. There were six pathological false-negative cases, a false-negative rate of 4.1%, all of which had only micrometastasis. Five cases received systemic adjuvant therapy and have been disease-free, however, one patient who refused further therapy developed infraclavicular lymph node metastasis two years after surgery. CONCLUSIONS: In the management of the patients with postoperatively revealed sentinel node micrometastasis, systemic adjuvant therapies might reduce local relapse without secondary lymph node dissection.

Abstract: Postoperative adjuvant chemotherapy reportedly improves advanced colorectal cancer patients' survival, however, it is necessary to assess what regimens are useful. Doxifluridine (5'-DFUR) is an intermediate of capecitabine approved in Europe and USA to treat metastatic colorectal cancer. 5'-DFUR is metabolized to 5-fluorouracil (5-FU) by thymidine phosphorylase existing in tumor at high concentrations, suggesting high 5-FU levels in tumor tissues and lesser complications. Present study compared usefulness of 5'-DFUR to that of oral 5-FU. Patients were enrolled at 38 centers from April 1993 to September 1996. They had diagnosed colorectal cancer of TNM stages II and III, and underwent macroscopic curative resection. Patients were prestratified into colon or rectum cancer and allocated into either 5'-DFUR (5'-DFUR 460 mg/m(2)/day + PSK 3 g/day) or 5-FU (5-FU 115 mg/m(2)/day + PSK 3 g/day) group by dynamic randomization (stratification factors such as depth of tumor, degree of lymph node metastasis, and location of tumor). Drugs were orally administered daily from postoperative week 2 to 54, with 6 mg/m(2) mitomycin C at operation and following days. Subjects for analysis were 277 in 5'-DFUR and 281 in 5-FU groups. Median follow-up was 6.5 years. Although no differences in overall survival curves were detected, multivariate analysis showed that 5'-DFUR + PSK regimen was a significantly better prognostic factor in patients with Dukes B or C (risk ratio, 1.451; p=0.048); with tumor depth of pT3 or pT4 (risk ratio, 1.568; p=0.020). For patients with advanced colorectal cancer, 5'-DFUR + PSK therapy may possibly be more useful than 5-FU + PSK, but further study is required.

Abstract: PURPOSE: Colon gas analysis using abdominal radiography has been reported as a reliable method for assessing functional bowel disorders. The aim of this study was to clarify the relevance of colon gas distribution in postoperative disorders such as constipation and feelings of incomplete evacuation following rectal cancer operation. METHODS: Colon gas volume score was calculated using plain abdominal radiographs and evaluated in 50 patients who had received low anterior resections. Twenty-one constipated patients who required laxatives and 29 patients who did not were compared in terms of colon gas distribution. In addition, 32 patients with postoperative feelings of incomplete evacuation and 18 patients without such feelings were assessed in similar fashion. RESULTS: Left colon gas scores in patients who required laxatives were significantly higher (2.82 +/- 3.23 percent) than in nonusers (1.21 +/- 0.96 percent; P < 0.01). Patients with feelings of incomplete evacuation displayed significantly higher left side colon gas scores (2.51 +/- 2.66 percent) than those without such feelings (0.77 +/- 0.81 percent; P < 0.0001). CONCLUSION: Patients with postoperative functional bowel disorders such as constipation or feelings of incomplete evacuation may experience relatively high volumes of gas in the left colon.

Abstract: BACKGROUND: Colon gas volume analysis using abdominal radiographs is an objective and reproducible method for evaluating functional bowel disorders. The aim of this study was to clarify the relevance of colon gas distribution and transit time in rectosigmoid cancer patients after surgery. METHODS: Segmental colon gas volume score was calculated using plain abdominal radiography and evaluated in 40 patients who had undergone sphincter-saving resection. Segmental colonic transit time was analysed using radiopaque markers in the same patients. RESULTS: Transit times in the right colon (RCT) were 15.3 +/- 1.1 h and in the left colon (LCT) 11.2 +/- 1.1 h. Gas volume scores in the RCS and LCS were 1.10 +/- 0.13% and 1.06 +/- 0.14%, respectively. Neither colonic transit time nor colon gas volume score correlated with the operation methods for rectosigmoid colon cancer. A positive correlation of RCS and a negative correlation of LCS/RCS with ageing were noted in male patients but not in female patients. There was no correlation between RCT and RCS (r = 0.028); however, LCT correlated with LCS (r = 0.318, P < 0.05). The ratio of colonic transit time (LCT/RCT) was 0.84 +/- 0.10, while that of colon gas volume score (LCS/RCS) was 1.29 +/- 0.21. There was a significant correlation between LCT/RCT and LCS/RCS (r = 0.541, P < 0.001). CONCLUSIONS: Analysis of colon gas volume is useful for evaluating colonic transit time in rectosigmoid cancer patients after sphincter-saving surgery.

Abstract: Many gastric cancer patients who recur peritoneally are initially diagnosed with serosal invasion. To clarify the usefulness of neoadjuvant chemotherapy with 5-fluorouracil (5-FU) +/- cisplatin (CDDP), neoadjuvant versus no preoperative chemotherapy for gastric cancer with preoperative serosal invasion was investigated. The patients were treated preoperatively with 5-FU 300 mg/m(2)/day for 2 weeks (F group; n=40), 5-FU 300 mg/m(2)/day for 2 weeks + CDDP 15 mg/m(2)/day for 2 days (FP group; n=80) or nothing (C group; n=100). A total of 78% of patients in C, 65.0% in F and 67.5% in FP group were classified as T3 or higher surgically. In patients without peritoneal metastasis, the positive peritoneal lavage cytology was 29.2% in C, 11.8% in F, and 12.2% in FP patients (p=0.0279). Serosal invasion was found histologically in 60.0% of C, 30.0% of F, and 33.8% of FP patients (p=0.001). There were no serious drug reactions and no increases in morbidity or mortality using either regimen. The 5-year survival rate was 47.0% in F and 50.9% in FP patients, but only 33.2% in C patients (p=0.0042). In conclusion, neoadjuvant chemotherapy with 5-FU +/- CDDP for gastric cancer patients with serosal invasion may reduce positive peritoneal cytology, eliminate cancer cells from the serosal surface, and improve prognosis.

Abstract: Doxifluridine (5'-DFUR) is an anticancer drug converted to 5-fluorouracil (5-FU) by thymidine phosphorylase (TP). TP is an angiogenetic and platelet-derived endothelial cell growth factor. We evaluated the relation between TP expression and chemotherapeutic efficacy and prognosis for gastric cancer. Advanced gastric cancer patients given oral adjuvant chemotherapeutics either 5'-DFUR; 163 patients or 5-FU; 162 patients were examined. TP expression was assessed with immunohistochemical staining. Multivariate analysis for influencing survival was done, employing variables such as gender, age, procedure, tumor size, location, Borrmann type, histologic factors [type, depth of invasion, lymph node metastasis (n), lymphatic invasion (ly), and venous invasion (v)], drug administered, and TP expression. In the patients with serosal invasion, 5'-DFUR in TP positive was an independent prognostic factor (risk ratio, 4.450; 95% confidence limit, 2.099-9.436), indicating significantly improved prognosis over the 5-FU group. In TP negative, n and ly were independent prognostic factors, but the survival curves of the two chemotherapeutic groups were not significantly different. TP expression was not prognostic factor in 5'-DFUR group, while, in 5-FU group, TP expression was an independent prognostic factor (2.834, 1.467-5.476). In conclusion, it was suggested that TP positive gastric cancer with serosal invasion increased malignant potential of the tumor and 5'-DFUR efficacy.

Abstract: BACKGROUND/AIMS: Local pelvic recurrence of rectal cancer after radical resection has been associated with morbidity and cancer-related death. This study retrospectively evaluated outcome following curative resection for rectal cancer recurring after surgery on the basis of prognosis, type of procedure and perioperative morbidity. METHODS: A total of 85 consecutive patients with local pelvic recurrence of rectal cancer were evaluated. Of these, 43 underwent microscopic curative surgery for local recurrence. Among the 43 patients, 23 underwent surgery alone and 17 received preoperative radiotherapy (40 Gy) (XRT group) in addition to the surgery. Of the 43 patients, 26 were asymptomatic. RESULTS: Curative resection was higher in the recurrences that were associated with implantation, incomplete surgical margin clearance, and intrapelvic lymph node metastasis than in other types of recurrence. With regard to surgical procedure, abdominoperineal resection (APR), with or without sacral resection, was standard following previous sphincter-preserving surgery, while total pelvic exenteration (TPE), with or without sacral resection, was common following previous APR. Local excision was not considered appropriate surgery. There was a high incidence of perioperative morbidity (64%) in patients receiving TPE. Re-recurrence was observed in 18 patients (50%) after curative surgery. After a follow-up of 2 years or more, the local re-recurrence rate was 28%. The overall 5-year survival rate for patients receiving curative resection was 39%, for patients in the XRT group, 51%, and for patients in the surgery-alone group, 24% (p = 0.07). The survival rate in 26 asymptomatic patients was higher than in 17 patients with symptoms, with 5-year survival rates of 62 and 23% (p < 0.05), respectively. The cumulative local control in the preoperative radiotherapy plus en bloc surgery group (XRT group) was significantly better than in the surgery-alone group (p < 0.01), and survival in the XRT group tended to be better than in surgery alone. CONCLUSIONS: These results suggest that careful patient selection according to the pattern of recurrence, area of invasion and presence of symptoms is important for successful curative surgery. Aggressive surgery with adjuvant therapy may lead to an improved salvage rate.

Abstract: OBJECTIVE: To clarify the usefulness of imprint cytology for intraoperative investigations of sentinel lymph nodes in breast cancer, comparing the results with those of examinations using frozen and permanent sections. STUDY DESIGN: The material consisted of 303 sentinel lymph nodes from 124 cases of clinically node negative breast cancer. Touch imprint cytologic slides and frozen sections were obtained from the same cut surface of the sentinel nodes. Correlations with the final histopathologic results in paraffin sections were evaluated. RESULTS: The sensitivity, specificity and accuracy of imprint cytology were 70.3%, 99.6% and 96.0%, and those of frozen sections were 83.8%, 100%, 98.0%, respectively. The values were improved when the 2 methods were combined (89.2%, 99.6%, 98.3%), though the concordance between imprint cytology and frozen section was 91.9%. CONCLUSION: Both imprint cytology and frozen section are useful for evaluating sentinel lymph node status in breast cancer. However, the 2 techniques should be combined to improve the diagnostic sensitivity.

Abstract: BACKGROUND: We investigated whether impaired preoperative natural killer (NK) cell activity correlates with asynchronous distant metastasis after curative surgery for rectal cancers. In addition, we examined if preoperative chemoradiotherapy for rectal cancers impairs NK cell activity and contributes to the induction of distant metastasis. STUDY DESIGN: Preoperative NK cell activity was examined in 174 rectal cancer patients. All patients were enrolled in this study and followed until asynchronous distant metastasis occurred. RESULTS: The mean NK activity in patients with stage IV disease (n = 20) was significantly lower than seen in other stages. There were no differences among stage I to stage III patients. In stage III patients, the cumulative distant metastasis-free rate after curative surgery was significantly lower in cases with NK activity of 25% or less than those with more than 25%. Preoperative chemoradiotherapy for stage I to III patients significantly impaired NK cell activity (n = 39), and the metastasis-positive ratio significantly increased among patients with stage II or stage III diseases (n = 30). Multivariate analysis indicated that dichotomized NK cell activity was a significant risk factor that is associated with distant metastasis as well as nodal involvement. CONCLUSIONS: In primary rectal cancers, NK cell activity is not necessarily impaired in accordance with the disease progression. It is considered an important background factor for developing asynchronous distant metastases in stage III rectal cancers. Neoadjuvant chemoradiotherapy impaired NK cell activity in selected patients, suggesting the necessity of concurrent immunotherapy for better outcomes.

Abstract:

Abstract: PURPOSE: SK1, a human IgM monoclonal antibody recognizes the antigen, termed AgSK1 which was shown to be preferentially expressed by human adenocarcinomas, particularly human gastrointestinal malignancies. The aim of this study was to clarify the clinicopathological significance of AgSK1 mRNA expression in human colorectal cancer. METHODS: Using a quantitative RT-PCR, we studied the mRNA expression level of AgSK1 in the resected specimen of 40 patients with colorectal cancer. We estimated the tumor tissue value (T-value), nontumorous tissue value (N-value), and the ratio of T-value to N-value (T/N Ratio). For further analysis, we compared these data with the clinicopathological features. RESULTS: In the status of lymph node metastases, pN positive status tended to have a higher T-value level of AgSK1 mRNA than pN negative status (P = 0.076). According to a mean T-value (8.032) or a mean T/N Ratio (1.549), we divided these patients into two groups, low expression group and high expression group. A high expression group showed a significantly higher frequency of positive lymph node metastases (T-value; P = 0.021, T/N Ratio; P = 0.024). CONCLUSION: AgSK1 mRNA expression in tumor tissue may become a useful marker for lymph node metastases and a malignant potential marker of colorectal cancer.

Abstract: BACKGROUND: Epstein-Barr virus (EBV), the etiological agent of infectious mononucleosis, has an important role in the oncogenesis of EBV-related malignant diseases. The association of EBV with gastric carcinoma cells has become well known recently, but there are only a few reports concerning its association with surrounding epithelia and infiltrating lymphocytes. In this study, we investigated the association of EBV with gastric carcinoma and surrounding cells. METHODS: One hundred and two cases of gastric carcinoma were studied. The specimens were studied for the presence of the EBV genome by polymerase chain reaction (PCR), and then by in situ hybridization (ISH) technique to determine the localization of EBV. RESULTS: Of 97 informative cases, EBV was detected in 21 cases (21.6%) by the PCR method. ISH studies showed that EBV RNA was expressed in 5 of the 97 cases (5.2%) and was localized to the nuclei of carcinoma cells. All these 5 lesions were found in male patients. In these 5 cases, 3 were diffuse type and 2 were intestinal type, and all cases arose in the proximal region of the stomach. EBV RNA was not detected in non-neoplastic epithelia, but it was detected in 24 of the 97 cases (24.7%) in small lymphocytes. CONCLUSION: EBV was detected in 5.2% of gastric carcinomas and in 24.7% of infiltrating lymphocytes by the ISH method. The high positive rate (21.6%) by the PCR method corresponds to the presence of the EBV genome in surrounding lymphocytes.

Abstract: BACKGROUND/AIMS: To determine whether preoperative natural killer (NK) cell activity has any prognostic significance in colon cancer patients. METHODS: The study population consisted of 140 patients with colon cancer. NK cell activity was determined within 2 weeks before surgery in 128 patients and at the time of diagnosis in the remaining 12 patients who either did not undergo surgery or who underwent palliative surgery only. Disease progression and postoperative prognosis were examined in relation to NK cell activity. RESULTS: Decreases in NK cell activity did not necessarily correspond to tumor stage. In curatively operated stage I-III diseases, preoperative NK cell activity of 20% or less correlated with poor survival. Lower activity was also associated with metachronous distant metastases but not with local recurrences. In particular, more than half the stage III patients with attenuated NK cell activity developed metastases. Multivariate analysis indicated that attenuated NK cell activity was a significant parameter for predicting distant metastasis following curative surgery for colon cancer. CONCLUSION: Preoperative NK cell activity has a significant prognostic value in curatively operated colon cancer, particularly for the development of metachronous distant metastasis in stage III patients.

Abstract: To evaluate the effect of neoadjuvant chemotherapy on gastric cancer, we examined the correlation between induction of apoptosis and expression of p53, Bcl-2, and Bax. Eighty-five patients with advanced gastric cancer were retrospectively divided into the following two groups: 54 patients received 5-fluorouracil (5-FU) at 300 mg/body/day for 14 days and cisplatin (CDDP) at 15 mg/body/day for 2 days as group A; 31 patients without any preoperative chemotherapy as group B. According to histological changes in tumors due to neoadjuvant chemotherapy, the therapeutic effects on tumors were evaluated. The apoptotic index (AI) of group A was significantly higher than that of group B (1.12+/-0.40 vs. 0.67+/-0.24; p<0.01). In group A, the AI of p53-positive cases was significantly lower than that of negative cases (0.92+/-0.32 vs. 1.39+/-0.32; p<0.01). The AI of histological responders was significantly higher than that of non-responders (1.34+/-0.35 vs. 1.02+/-0.38; p<0.01). There was no significant correlation between AI and expression of Bcl-2 or Bax. In group A, histological responders, Bcl-2 positive, and high AI patients had better prognosis, respectively. In conclusion, neoadjuvant chemotherapy for gastric cancer enhanced induction of apoptosis, and AI might be useful to evaluate the effect of neoadjuvant chemotherapy.

Abstract: Dihydropyrimidine dehydrogenase (DPD) and dihydropyrimidinase (DHP) are metabolic enzymes of fluoropyrimidine. UFT containing uracil (U) and Tegafur is the first reported DPD-inhibitory fluoropyrimidine. To clarify the significance of the enzyme activities, we examined the relationships between the effects and adverse reactions, and DPD and DHP activities in gastric cancer treated with UFT1+cisplatin neoadjuvant chemotherapy. Twenty-five gastric cancer patients were administered UFT at 370 mg/m(2)/day for 21 days and cisplatin at 15 mg/m(2)/day for 2 days. Dihydrouracil (DU) and U levels in the urine and DPD activities in the resected tumors were measured. Chemotherapeutic effects were classified histologically into non-responder and responder groups. The responder group accounted for 48% of the patients. All six patients with high DPD activities (> or = 0.08 nmol/min/ww) belonged to the non-responder group and 11 of 19 patients with low DPD activities (<0.08 nmol/min/ww) belonged to the responder group; the difference was significant (p=0.0435). Adverse reactions to UFT occurred in four patients, all of whom were among the six patients with abnormal DU/U values. The incidence of UFT adverse reactions was estimated at 67%. In conclusion, the measured levels of DPD-related enzyme activities appear to be significant for predicting the effects and adverse reactions to chemotherapy.

Abstract:

Abstract: Chemoradiotherapy for the treatment of primary rectal cancers has been regarded as an effective adjuvant for surgical procedures. It has been expected to reduce local recurrence, increase the chance of sphincter-preserving operation and improve survival. However, recent advances in surgical technique for rectal cancers have reduced the local recurrence rate to no more than 10%, where the question may arise of whether adjuvant chemoradiotherapy is indeed necessary. Another matter to be elucidated is why adjuvant chemoradiotherapy does not necessarily improve overall survival for rectal cancers even when the local recurrence rate is reduced. In the treatment of locally recurrent lesions, either heavy ion radiotherapy or proton beam therapy is expected to be an effective alternative to the surgical procedure.

Abstract: The clinically useful monoclonal antibodies(Mabs) for colorectal cancers were reviewed. Since 1980's, immunoscintigraphy has been performed for the detection of occult colorectal cancers. However, it may be substituted with the development of positron emission tomography. As for the treatment, some Mabs have been shown to be effective for the adjuvant therapy of postoperative colorectal cancers. Some Mabs to epidermal growth factor receptors(EGFr) are quite promising since they block the functions necessary for the tumor growth and enhance the cytotoxicity of chemotherapy. Recent advances for the development of humanized Mabs will improve the chance of Mabs to be used as an effective adjuvant.

Abstract: Recent molecular biological studies suggested certain molecular markers might be useful as prognostic factors in patients with colorectal cancer. The purpose of this study was to investigate whether cyclin dependent inhibitor kinase p21WAF1/CIP1 (p21), p53 expression, and/or the presence of apoptosis had prognostic value in predicting survival in patients with advanced middle and lower rectal cancer who were treated with preoperative radio-chemotherapy. We examined the immunohistochemical expression of p21 and p53, and determined the degree of apoptosis in resected middle and lower rectal cancers from patients who received preoperative radio-chemotherapy (irradiation group, n=40) and from those who did not receive treatment (control group, n=35). The preoperative total radiotherapy dose was 42.6 Gy and the chemotherapy tegafur suppository dose was 750 mg/day. Clinicopathological features, and tumor expression of p53 and p21 and degree of apoptosis were analyzed by means of multivariate analysis. In the irradiation group, tumors were positive for p53, p21 and apoptosis in 34 of 40 (85.0%), 23 of 40 (57.5%) and 25 of 40 (62.5%) cases, respectively. The expression of p21 and the apoptotic index were significantly higher in the irradiated group compared to controls (2.0 versus 1.2%, p=0.05; 8 versus 3%, p=0.03, respectively). There was a significant correlation between p21 immunoreactivity and the degree of muscularis propria invasion (p=0.004), as well as between p21 immunoreactivity and survival rate (p=0.03). Multivariate analysis revealed that p21 expression (RR, 0.09; 95% CI, 0.01-0.78; p=0.03) and lymph node metastasis (RR, 3.63; 95% CI, 1.06-12.37; p=0.04) were significant prognostic factors for patient survival. These data suggested that p21 expression has prognostic value in predicting patient survival in advanced middle and lower rectal cancer.

Abstract: Adhesion molecules can initiate intracellular signaling. Engagement of CD44 either by its natural ligand hyaluronan or a specific antibody on a cell line induced tyrosine phosphorylation and activation of focal adhesion kinase (FAK), which then associated with phosphatidylinositol 3-kinase (PI3K) and activated mitogen-activated protein kinase at its downstream. However, the introduction of dominant negative Rho into the cells inhibited the CD44-stimulated FAK phosphorylation. Cells expressing CD44 were significantly resistant to etoposide-induced apoptosis. This anti-apoptotic effect was cancelled by the inhibition of either Rho, FAK or PI3K. These results may indicate a signaling pathway from CD44 to mediate the resistance against drug-induced apoptosis in cancer cells.

Abstract: BACKGROUND: Total pelvic exenteration (TPE) for the treatment of advanced colorectal cancer usually involves a double stoma for faecal and urinary excretion, which reduces patient quality of life. In this study, a stomaless reconstruction method for patients normally requiring TPE was evaluated. METHODS: Five patients underwent stomaless TPE. After removal of the tumour with an adequate surgical margin, the urethra was transected at the urogenital diaphragm and the rectum at the anal canal. An ileal neobladder was constructed and coloanal anastomosis was performed. The major omentum was used to construct a septum between the anastomoses. A transgastric ileus tube was used as an intestinal stent to prevent ileus. RESULTS: All patients were alive 12-39 months after operation. Faecal continence was preserved in four patients whose diverting colostomies were closed. All five patients were able to void urine spontaneously, with daytime continence. All but one, in whom cancer recurred, were mobile in the community. CONCLUSION: Stomaless TPE may be considered for locally advanced colorectal cancers that invade the genitourinary organs, provided that neither the anal canal nor the urogenital diaphragm is affected.

Abstract: Metabolic bone disorders are recognized as one of the late complications after gastrectomy. However, the onset time and the extent of bone disorders are still unknown. We examined the influence of active vitamin D treatment on bone metabolism in the early period after gastrectomy. Sixty-three postgastrectomy patients were divided into two groups; active vitamin D treatment group [VD(+)] and no treatment group [VD(-)]. The level of serum calcium and phosphate was increased in the VD(+) group compared with the preoperative level, and parathyroid hormone (PTH-M) was decreased in the VD(+) group. Both 1,25-(OH)2D3 and bone-specific alkaline phosphatase (B-ALP) were increased in the VD(-) group. Cross-linked carboxyterminal telopeptide of type I collagen (I-CTP) was increased in the VD(+) group. There was no change in calcitonin in either group. In conclusion, metabolic bone disorders after gastrectomy have their onset in the early period, and active vitamin D treatment from the early period may be effective in preventing bone disorders.

Abstract: BACKGROUND: Using computerized nuclear morphometry, we searched for common, objective parameters for use in predicting potential malignancy of primary T2 tumors of the rectum. METHODS: In 119 surgically resected T2 rectal cancers, the following parameters of the cancer cell nuclei were microscopically measured: mean nuclear area (NA), mean perimeter (PM), largest-to-smallest diameter ratios (LS), circularity factor (CF), and coefficient of variation of nuclear area (NACV). The calculated parameters were retrospectively analyzed and compared with patients' outcome. RESULTS: NA, PM, and NACV correlated with recurrence. NA and NACV were significantly associated with disease-free survival. High NACV correlated with lymph node metastasis. In node-negative cases, high NACV correlated with recurrence and tended to be associated with poor prognosis. CONCLUSIONS: These findings suggest that computerized morphometry is useful for predicting malignancy potential of T2 lesions of the rectum.

Abstract: There are few reports on overall usefulness of adjuvant chemotherapy in gastric cancer patients. We tried to clarify, using multivariate analysis, usefulness of postoperative adjuvant oral chemotherapy in advanced gastric cancer patients after curative resection. Four hundred and eighty-two gastric cancer patients enrolled in a randomized controlled trial were classified into 2 groups based on postoperative chemotherapeutic regimen: oral doxifluridine (5'-DFUR, an intermediate metabolite of capecitabine) (n=245) or oral 5-fluorouracil (5-FU) (n=237). The significant prognostic factors in patients with serosal invasion were chemotherapeutics (5'-DFUR vs. 5-FU) (risk ratio 1.649; 95% CI, 1.112-2.437), lymph node metastasis (no vs. yes) (2.823; 1.422-5.604), and tumor differentiation (differentiated vs. undifferentiated) (1.727; 1.068-2.791). Significant factors influencing peritoneal recurrence time were chemotherapeutics (1.756; 1.063-2.902), serosal invasion (no vs. yes) (2.237; 1.264-3.961), lymph node metastasis (2.541; 1.267-5.095), tumor differentiation (2.656; 1.374-5.136), and tumor location (others vs. total) (3.595; 2.006-6.443). There were no differences in the overall survival between chemotherapy. However, 5'-DFUR produced a better survival time of patients with serosal invasion than 5-FU, that might be attributed to the prevention of peritoneal recurrence in this subset.

Abstract: BACKGROUND: The human monoclonal antibody SK-1 recognizes a glycoprotein expressed on the majority of colon cancer tissues. In the current study, we evaluated the safety, toxicity and preliminary efficacy of escalating dosages of SK-1 in patients with advanced colon cancer. PATIENTS AND METHODS: SK-1 was administered intravenously at 2, 4 or 10 mg three times to three groups of patients with recurrent colon cancer. The clinical outcome and the induction of serum anti-idiotypic antibody (Ab2) were assessed periodically. RESULTS: The mean rate of serum CEA level increase declined significantly during the eight weeks following the treatment. In four patients, serum titer of anti-idiotypic IgG antibodies to SK-1 (Ab2) continued to increase following the treatment. CONCLUSION: HuMAb SK-1 was well-tolerated and can be safely administered. It was suggested that SK-1 natural antibody not only possessed direct cytostatic activity against colon carcinoma, but may also have induced carcinoma-related, anti-idiotypic antibody responses.

Abstract: BACKGROUND/AIMS: The role of liver resection for hepatic metastases from noncolorectal carcinomas has yet to be clarified. The present study examines a single institutional experience of hepatic resection for noncolorectal metastases. METHODOLOGY: From January 1987 to March 1999, 14 patients underwent curative resection for liver metastases from noncolorectal carcinomas. Records of these patients were reviewed. RESULTS: Resections were performed for liver metastases from gastric cancers (n = 8), pancreatic cancers (n = 2), and cancers of bile duct, the papilla of Vater, kidney, and breast (n = 1, each). Six patients (5 with gastric cancers and 1 with pancreas cancer) presented with synchronous disease and 8 with metachronous disease. In the gastric cancer patients, there are 2 disease-free survivors (26 and 53 months) in the metachronous group, though all of the 5 patients with synchronous disease died within 29 months. All of the 4 patients with pancreatobiliary carcinomas died within 2 years. One case of breast cancer and another of renal cell cancer are alive without disease at 49 and 9 months, respectively. CONCLUSIONS: For metastases from gastric cancers, better survival after hepatic resection is expected in metachronous cases than in synchronous cases. Hepatic resection may afford little benefit for patients with liver metastases from pancretobiliary cancers.

Abstract: Thymidine phosphorylase (TP) is an enzyme which converts doxifluridine (5'-DFUR) to 5-fluorouracil (5-FU). To assess whether TP expression is useful for selecting adjuvant chemotherapy in advanced gastric cancer, we compared effects of oral 5'-DFUR and 5-FU and assessed correlation between drug efficacy and TP expression level. We examined TP expression in 286 patients. TP expression was assessed with immunohistochemical staining. When we compared prognosis in two chemotherapy groups with high TP expression, better survival was observed in 5'-DFUR than in 5-FU group (p=0.0413). Especially in stage III, patients with high TP had better survival in 5'-DFUR than in 5-FU group.

Abstract: SK1, a human immunoglobulin M (IgM) monoclonal antibody was derived from regional nodal lymphocytes of a Dukes B colon carcinoma patient. The antigen recognized by the human monoclonal antibody (HuMab) SK1, termed AgSK1, was shown to be a two-chain glycoprotein with an apparent molecular weight range of 42-46 kDa and preferentially expressed by human adenocarcinomas, particularly human gastrointestinal malignancies. To identify the gene encoding the AgSK1 antigenic epitope, a cDNA expression library constructed in lambda gt22A using mRNA from the colon carcinoma cell line HT29 was screened and one of the isolated clones encoding a 1.5-kb cDNA, which showed strong immunoreactivity with HuMab SK1, was selected for further analysis. This clone consisted of an amino terminal open reading frame of 54 amino acids and the carboxyl terminal 20 amino acids of this protein coding region contained the antigenic epitope recognized by HuMab SK1.

Abstract: Since 1985, we have studied autonomic nerve-sparing surgery combined with preoperative radiochemotherapy in patients with advanced rectal cancer to preserve genitourinary function without compromising radicality. The aim of this study was to evaluate postoperative genitourinary function and prognosis. A total of 84 patients with advanced rectal cancer underwent curative surgery combined with preoperative radio-chemotherapy. Of them, 76 underwent curative nerve-sparing surgery using radiation (42.6 Gy) and tegafur suppository. Postoperative genitourinary function, survival, and pelvic recurrence were investigated in these patients. The 5-year and 10-year disease-free survival rate was 80.7%, and the pelvic recurrence rate was 7.9% in patients receiving curative nerve-sparing surgery combined with radio-chemotherapy. All of these patients could micturate spontaneously, but preservation of sexual function was not as successful. Although this combined therapy is useful for curability and good postoperative quality of life, a new approach is needed to preserve sexual function.

Abstract: Lung cancer is the leading cause of cancer deaths; patients with this disease often develop antibodies to their own tumour antigens. TB94 is one such human antibody, and was obtained from the natural immune response from a patient with lung cancer. The data generated so far on TB94 suggest that this human antibody may have clinical applications in the diagnosis and treatment of lung cancer. Since humans make antibodies to tumour antigens, the intelligence of the human immune response can be exploited for the discovery of potential novel antigens. These novel antigens can then be developed for vaccine applications. Creating effective strategies to analyse systematically the natural human immune response to tumour antigens will open up new areas of immunotherapy for the control and eradication of disease.

Abstract: BACKGROUND: The clinical value of positron emission tomography (PET) for the diagnosis of local pelvic recurrence of colorectal cancer was evaluated. METHODS: Computed tomography (CT) and magnetic resonance imaging (MRI) of the pelvis were performed at regular intervals in 23 patients who had undergone resection for colorectal cancer. The 23 patients had a total of 25 lesions. PET images of the 25 lesions and of six primary lesions in patients with rectal cancer were obtained. A differential absorption ratio (DAR) was calculated in order to examine the accumulation of [18F]2-fluoro-2-deoxy-D-glucose (18FDG) on PET images. Histological diagnoses of the pelvic masses were obtained by CT-guided needle biopsy. RESULTS: On CT or MRI, a pelvic mass with a spicular shape (n = 1) was non-recurrent, whereas a nodular or lumpy shape indicated a locally recurrent lesion (n = 10). Masses with a nodulospicular shape (n = 12) did not correlate with the histological features. On PET, 15 of 16 histologically proven local recurrences were imaged positively. By setting a DAR of 2.8 as a cut-off value, local recurrences could be diagnosed with 100 per cent accuracy. CONCLUSION: PET is a clinically useful tool for the detection of local recurrence of colorectal cancer, particularly for distinguishing between recurrence and granulation tissues in the pelvic cavity.

Abstract: Several nerve-sparing operations for advanced rectal cancer that aim to preserve genitourinary function without compromising tumor clearance have been developed in Japan. The aim of this study was to evaluate the survival and local recurrence of these procedures in Dukes B and C patients. A total of 177 patients with advanced rectal cancer underwent curative nerve-sparing surgery (NSS) over the last 11 years; 52 were Dukes B patients and 54 were Dukes C. Altogether 36 had Dukes C1 and 18 had Dukes C2 tumors, 13 with lateral lymph node metastases, designated lateral LN(+). The 5-year survival rate was 92% for Dukes B, 67% for Dukes C1, and 39% for Dukes C2 patients: 11% for Dukes C2 patients with lateral LN(+). The local recurrence rate was 6% for Dukes B, 11% for Dukes C1, and 33% for Dukes C2 patients: 20% for the lateral LN(-) group and 39% for the lateral LN(+) group. Almost all of the patients undergoing NSS could micturate spontaneously, but preservation of sexual function was not as successful. Although there is no guarantee of preserving satisfactory sexual function, our NSS is an acceptable procedure for Dukes B, C1, and C2 patients without lateral lymph node metastases.

Abstract: BACKGROUND AND OBJECTIVES: We evaluated the clinical efficacy of adoptive immunotherapy using in vitro activated cytotoxic T lymphocytes (CTL) in the treatment of patients with advanced cancer. METHODS: CTL were induced with the mixed lymphocyte and tumor cell culture method, in which lymphocytes isolated from patient peripheral blood mononuclear cells were mixed with inactivated autologous tumor cells. Activated lymphocytes were administered intravenously to 11 patients once every 2 weeks for 10 weeks (i.e., 5 doses). RESULTS: Tumor reduction and decreased tumor marker were observed in 4 patients. Notably, successful CTL induction was identified in all of these patients. In patients who did not show induction of CTL response, a decreased proportion of lymphocytes, especially CD8(+) cells, and increased levels of CD14(+) cells were frequently observed. Fluorescence-activated cell sorter analysis indicated that expression of HLA class I and costimulatory factor B7-1 molecules was diminished on tumor cells. This was partly recovered with interferon-gamma, which resulted in successful induction of a CTL response. CONCLUSIONS: It was suggested that in vitro CTL induction is difficult in patients with advanced cancer. However, once the cells were induced successfully, some favorable clinical effects were seen by the adoptive transfer of such cell populations.

Abstract: This retrospective study evaluated outcome with regard to procedure, local control, and survival after curative surgical resection with and without preoperative radiotherapy for local pelvic recurrence. A total of 58 consecutive patients with local pelvic recurrence of rectal cancer after previous curative resection for primary tumors were reviewed. Of these, 36 underwent both initial resection and follow-up in our department; the remaining 22 had initial surgery and follow-up elsewhere. Of the 58 patients 27 underwent curative re-resection, 9 had palliative resection, and 22 were treated by conservative therapy. Among the 27 patients with curative resection 17 received preoperative radiotherapy (40 Gy) plus surgery and 10 surgery only. No patients were lost to follow-up; median follow-up time was 36.3 months. The overall rate of curative resection was 46.6%: 55.6% in our own follow-up group and 31.8% in the others. With regard to surgical procedure, abdominoperineal resection (APR) with or without sacral resection was standard following previous low anterior resection, and total pelvic exenteration (TPE) with or without sacral resection was common following APR. There was a high incidence of morbidity (71.4%) after TPE. Re-recurrence was observed in 12 (44.4%) after curative re-resection. There was local re-recurrence in 6 (22.2%). The local re-recurrence rate was 11.8% (n = 2) with radiotherapy plus surgery, and 40.0% (n = 4) with surgery alone. The estimated 5-year survival following curative re-resection was 45.6% (61.2% with radiotherapy plus surgery, 29.6% with surgery alone). Both survival and local control with radiotherapy plus surgery tended to be better than with surgery alone. Thus, in selected patients pelvic local recurrence of rectal cancer can be re-resected curably by APR or TPE (with or without sacral resection) combined with preoperative radiotherapy.

Abstract: BACKGROUND: The current study was undertaken to evaluate the quantitative estimation of cytologic features on aspirated smears for the preoperative differential diagnosis of follicular lesions of the thyroid. METHODS: The subjects were 60 patients with follicular lesions of the thyroid (including 20 follicular carcinomas, 15 follicular adenomas, and 25 adenomatous goiters) whose histopathologic explorations were conducted fully postoperatively. Using a microscope connected to a computerized video system, the mean nuclear area, the mean nuclear perimeter, the circular rate, the largest to the smallest dimension ratio (LS ratio) of the nuclei, and the coefficient of variation of the nuclear area (NACV) were measured and analyzed. RESULTS: Among the quantitative morphometric parameters of nuclei, the circular rate was significantly higher in the group with adenomatous goiters than those with follicular carcinomas (P < 0.00001) and adenomas (P < 0.005). The group with follicular carcinomas had a higher LS ratio than the group with adenomatous goiters (P < 0.0005). The NACV value increased as the malignant potential of the lesion increased and showed significant differences between the groups. When a NACV of 21.5% was chosen as the cutoff point, the incidence of malignancy was significantly higher in patients with high NACV values than in those with low NACV values (P < 0.00001). Using this borderline value, it was possible to distinguish malignant from benign diseases with a sensitivity of 85.0%, a specificity of 82.5%, and an accuracy of 83.3%. CONCLUSIONS: Preoperative quantitative estimations of cytologic nuclear features are useful for the preoperative differential diagnosis of follicular lesions of the thyroid.

Abstract: We characterized a natural human antibody to adenocarcinomas and investigated the biological role of this Ab/Ag complex in cancer expansion. Human monoclonal antibodies (HuMAbs) were generated with hybridoma fusion methods using regional nodal lymphocytes of colon carcinoma patients. Among 1036 HuMAbs, only one, termed SK1, an IgM, was adenocarcinoma specific in the immunohistochemical study. The antigen recognized by SK1 (Ag-SK1) was a glycoprotein with a molecular weight of 42-46 kDa. The expression of Ag-SK1 on carcinoma cells varied according to the cell growth periods but was independent of cell cycle state as elucidated by two-colour fluorescence-activated cell sorter (FACS) analysis. A dot-blot analysis showed that the concentration of Ag-SK1 per total protein differed considerably among eight colon carcinoma cells examined and that the difference was closely correlated with the invasion capacity of the cells as assessed by a microchemotaxis assay. Furthermore, up to 87% of cell migration was inhibited by SK1 in a dose-dependent manner. These data suggested that Ag-SK1 is metabolized and expressed on highly invasive carcinoma cells. In addition, it appears that, although rare, some patients do mount an anti-cancer antigen response in their draining lymph nodes. A HuMAb such as SK1 may be a good candidate for the treatment of cancer invasion and metastasis.

Abstract: BACKGROUND: Since 1984, we have studied nerve-sparing surgery (NSS) combined with preoperative radiotherapy (XRT) in patients with advanced rectal cancer to preserve the genitourinary function without compromising radicality. The present aim was to evaluate the prognosis and the postoperative genitourinary function. METHODS: A total of 167 patients with advanced rectal cancer underwent curative nerve-sparing surgery. Among them, 60 underwent the preoperative therapy using irradiation (42.6 Gy) and tegafur suppository. Survival, local recurrence, and postoperative genitourinary function were investigated in these patients. RESULTS: The 5-year survival rate was 80.9% and the local recurrence rate was 6.7% in the NSS+XRT group. Almost all of the patients receiving NSS could micturate spontaneously, but preservation of sexual function was not as successful. CONCLUSIONS: Better local control and preservation of urinary function were possible in advanced rectal cancer patients by NSS+XRT.

Abstract: The authors investigated whether host immunity contributes to the development of asynchronous distant metastases in colorectal carcinomas. The host immunity was examined 8 times, pre- and postoperatively during a one year period in 77 curatively operated cases. A prospective study was performed using obtained personal data. During the mean follow-up period of 920 days, 13 patients developed distant metastases. Among the immunological parameters, the preoperative natural killer (NK) cell activity differed significantly between the metastases positive and negative groups. On univariate analysis, dichotomous NK activity, presence of nodal metastases, and venous invasion correlated with metastases. The hazard ratios on multivariate analysis were 4.53, 3.82, and 4.81, respectively. No correlation was noted between NK activity and the progression stages of colorectal carcinomas. These data suggested that attenuated preoperative NK activity is an important background factor for the development of asynchronous distant metastases following curative resection of colorectal carcinomas.

Abstract:

Abstract: BACKGROUND: Nonhuman monoclonal antibodies (MoAbs) of desired specificities have been studied in cancer treatment and tumor targeting with minimal success. Attempts of using humanized chimeric antibodies have not improved significantly their clinical applications. We have engaged in the development of human MoAbs by incorporating the in vitro immunization protocols to the nodal lymphocytes of cancer patients. Three human MoAbs thus generated were found to be strongly reactive with various human malignancies. The antigens recognized by the three antibodies were selected for immunochemical and biochemical characterizations. METHODS: The antigens investigated were AgSK1, PA 1-2 and PA 3-1. The patterns of each antigen expression in various human cancer cell lines were studied by the immunocytochemical staining technique. The expression of AgSK1 in association with cellular proliferation was examined by the flow cytometry analysis. In studying the biochemical natures of these antigens, their sensitivities toward various chemical and physical treatments were determined. The antigens that were shown to be proteins were subjected to SDS-PAGE and Western blot for estimations of molecular weights. RESULTS: The AgSK1 was detected in 10 human carcinoma cell lines but in none of the melanoma cell lines. This suggests that SK1 may be an epithelial or carcinoma marker. The phenotypic expressions of AgSK1 were shown to be associated with proliferation of carcinoma cells. Biochemically AgSK1 was a sialophycoprotein with an estimated molecular weight of 42-44 kilodaltons (kDa). HuMAb PA1-2 demonstrated a unique staining pattern at both the cytoplasmic and intercellular interface. The stained filamentlike structures extending from cell to cell indicated that Ag PA1-2 might play a role in cellular interactions. Biochemically, Ag PA1-2 appeared to be an asialocarbohydrate. The Ag PA3-1 was a cytoplasmic glycoprotein expressed by all 13 cell lines. The estimated molecular weights of PA3-1 were 164, 104, and 40 kDa. CONCLUSIONS: Tumor-associated antigens recognized by the human MoAbs may be more relevant clinically than those recognized by the mouse immune system. Carcinoma-specific human MoAbs are desirable for cancer treatment and tumor localization.

Abstract:

Abstract: An IgM human monoclonal antibody (HuMAb) SK1 was generated from mesenteric nodal lymphocytes of a colon cancer patient that were fused with a human B-lymphoblastoid cell line SHFP-1. The reactivities of HuMAb SK1 to various human cell lines were screened by cell enzyme linked immunosorbent assay and immunocytochemical staining. The HuMAb SK1 reacted strongly with all 11 human carcinoma cell lines that were tested and had no detectable binding with noncarcinoma cell lines of the following origins: fibroblast; fetal lung; melanoma; soft tissue sarcoma; neuroblastoma; and glioblastoma. Carcinoma preferred reactivity of HuMAb SK1 was further confirmed by immunoperoxidase staining of a large number of frozen tissues, both malignant and benign. The antigen SK1 (AgSK1) in human carcinoma detected by immunoperoxidase staining was also identified biochemically as a sialoglycoprotein that migrated at M(r) 42,000 with an isoelectric point (pI) of approximately 5.9. A preferential staining by HuMAb SK1 was seen among colorectal, gastric, pancreatic, and lung cancers. Competitive inhibition study in solid-phase immunoassay suggested that the HuMAb SK1 did not cross-react with other antibodies specific for CEA, CA 19-9, and TAG 72. The AgSK1 appears to be a novel carcinoma associated antigen which may be a useful tumor marker in cancer diagnosis and treatment.

Abstract: A delay in red cell recovery after ABO-incompatible bone marrow transplantation (BMT) is often observed. The authors experienced a case of prolonged anemia after a major ABO incompatible BMT for myelodysplastic syndrome which was successfully treated with recombinant human erythropoietin (Epo). Effects of Epo were confirmed by the recurrence of anemia after withdrawal of Epo as well as the rapid reincrease in reticulocytes on readministration. The patient received a dose of Epo which was similar to the amount used for renal anemia, however serum concentration of Epo after administration exceeded endogenous Epo levels. Epo may have a beneficial role in the treatment of prolonged anemia after BMT.

Abstract: The use of the severe combined immunodeficient mouse (SCID), CB-17/Icr//Imd-SCID, was investigated for the production of human hybridoma ascites containing human antibody. Human-human hybridomas, generated from the fusion of lymphocytes isolated from regional draining lymph nodes of cancer patients with the SHFP-1 fusion partner, were injected i.p. at various cell concentrations into pristane-primed SCID mice. Ascites growth was typically observed at 7-14 days postinoculation. No significant differences in ascites yield or production were observed between IgG- and IgM-secreting hybridomas. Yields of immunoreactive human immunoglobulin ranged from approximately 0.5 to 3 mg/ml of harvested ascites. The ease and relatively low cost suggest that the use of SCID mice is preferred over conventional and costly large-scale industrial procedures.

Abstract:

Abstract: Lymphocytes from regional lymph nodes of patients with colon cancer were fused with a human lymphoblastoid cell line with or without in vitro immunization. The efficacy of these two protocols for the generation of human monoclonal antibodies against colon cancer was investigated. The hyperplastic lymph nodes adjacent to the tumor were the best source of B lymphocytes. Fusion frequency and the number of tumor-reactive clones were markedly increased when the in vitro immunization protocol was applied prior to fusion. As a stimulant in in vitro immunization, the supernatant of pokeweed mitogen-stimulated T lymphocytes was superior to the supernatant of mixed lymphocytes culture. Carcinoembryonic antigen at 20 micrograms/L seemed to be the optimal dose for in vitro immunization. The reactivities of human monoclonal antibodies thus generated were measured by enzyme-linked immunosorbent assay and confirmed by immunoperoxidase staining. Combining in vitro immunization with lymphocytes of cancer patients may lead to the successful production of clinically useful human monoclonal antibodies.

Abstract: Human lymph node lymphocytes from cancer patients were fused with either the UC 729-6 or SHFP-1 human fusion partners. Resulting human-human hybridomas were tetraploid, expressed markers from both parent cells, and secreted approximately 1 microgram Ig/10(6) cells/ml/day. Immunofluorescence analysis of some of the human MAbs with a panel of normal and malignant cell lines revealed a staining pattern of only the nuclear region. One IgM secreting hybridoma, TLN1F4, derived from a teratocarcinoma lymph node, predominantly stained the nuclear regions of adherent tumor cell lines and no hematopoietic cell lines or normal fibroblasts. PLN3C8, an IgG1 secreting hybridoma, derived from a prostate carcinoma lymph node, predominantly stained the nucleolus of LnCap, a a carcinoma of the prostate cell line. CLN2E5, an IgM secreting human hybridoma, derived from a carcinoma of the cervix lymph node, predominantly stained both cytoplasmic and nuclear components to tumor cell lines and not normal fibroblasts or hematopoietic cell lines. These data suggest that the immune response occurring within regional draining lymph nodes is capable of recognizing nuclear-associated antigens.