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The octacosapeptide amide corresponding to the entire amino acid sequence of chicken VIP was synthesized in a conventional manner, using a new arginine derivative, NG-mesitylene-2-sulfonylarginine, Arg(Mts). Treatment of a fragment, Z(OMe)-Thr-Asp-Asn-Tyr-NHNH2 with methanesulfonic acid or HBr was found to give a product with a low recovery of Asp, after aminopeptidase digestion. Ring closure of the Asp-Asn unit seemed to be responsible for this phenomenon. Deprotection with HF or TFA exhibited definitely less such a tendency. In the final step of the synthesis, all protecting groups, including the Mts group, were removed by HF in the presence of m-cresol and the deprotected peptide was purified by ion-exchange chromatography on CM-cellulose followed by isoelectric focusing in Ampholine pH 9--1. Synthetic peptide exhibited the identical Rf value with that of natural chicken VIP and was active as the natural peptide.

Background. Great progress in the diagnosis and surgical treatment of hepatocellular carcinoma (HCC) has led to an increased number of resectable cases. Much attention has been paid to the factors affecting long term survival of patients with HCC after partial hepatectomy. Methods. Survival rates for approximately 5800 patients with HCC registered by the Liver Cancer Study Group of Japan who received partial hepatic resection from January 1, 1982 to December 31, 1989 were calculated relative to 14 clinicopathologic variables. Cox's multivariate analysis and the stepwise method then were performed to determine significant prognostic variables and to investigate the appropriate combination of these variables for predicting prognosis. Results. The use of Cox's multivariate proportional hazard model indicated that significant prognostic indicators were alpha-fetoprotein level, tumor size, number of tumors, accompanying cirrhosis, age, surgical curability, and portal involvement. The stepwise method showed that the most valuable prognostic factor was portal involvement, followed by number of tumors, alphafetoprotein level, tumor size, accompanying cirrhosis, age, and surgical curability. Furthermore, it was found that the best predictive combination of two factors was number of tumors and alpha-fetoprotein level and that the best predictive combination of three factors was the alpha-fetoprotein level, tumor size, and number of tumors. Conclusion. This extensive analysis provides information that can be used to predict the prognosis of patients with HCC after undergoing partial hepatectomy.

We report herein the cases of two Japanese patients with typical fibrolamellar hepatocellular carcinoma treated at our institute. The first patient was a 19-year-old man with no hepatitis B or C viral infection and a normal, noncirrhotic liver in the nontumorous area. The second was a 36-year-old woman with no viral infection and a noncirrhotic liver in the nontumorous area. The clinicopathology, imaging appearances, and histology of both cases were similar to reports from the United States.

To assess the prognostic value of DNA ploidy in colorectal cancer, compared with the histopathological findings, paraffin-embedded surgical specimens from 330 patients who underwent resection for primary adenocarcinoma were studied using a new modified method of flow cytometry. Of these specimens, 141 were DNA diploid and 189, DNA aneuploid, among which there were 3 DNA hypodiploid lesions. Of the ten variables studied in curative resection, DNA ploidy ranked fourth in prognostic significance according to the linear trend by the chi 2 test, after nodal status, grade of cellular differentiation, and degree of invasive growth, if the DNA ploidy pattern was classified into three categories. Conversely, DNA ploidy was the sixth most significant factor if DNA hypodiploidy was included in the DNA aneuploidy. The Cox multivariate analysis showed that DNA ploidy was one of the five significant factors independently determining prognosis; however, if adjustment for the modified Dukes' stage was made by the Mantel-Haenszel test, the survival difference between the diploid and aneuploid groups did not reach a statistically significant level. Thus, we conclude that from a practical point of view, DNA ploidy is not an essential factor which must be combined with histopathological variables for a better prediction of patient outcome.

To study the role of pancreatic blood flow and vasoactive substances in the development of acute pancreatitis, we measured portal vein blood levels of bradykinin, prostaglandin E2 (PGE2), histamine, serotonin, and pancreatic enzymes, and with an electromagnetic blood flowmeter we recorded gastroduodenal arterial flow (GDAF), superior mesenteric arterial flow (SMAF), and mean arterial blood pressure for 6 hr in dogs with acute hemorrhagic necrotizing pancreatitis induced by the retrograde injection of autologous bile (0.5 ml/kg) into the pancreatic duct. GDAF and SMAF decreased immediately in the early phase of acute pancreatitis (-17.8 +/- 6.1%** at 10 min and -15.8 +/- 7.1%* at 20 min; *P < 0.05, **P < 0.01); portal bradykinin concentration increased quickly (3.2 +/- 1.2 pM at 0 time, 16.2 +/- 5.2 pM* at 5 min, 30.4 +/- 4.8** pM** at 10 min, and 39.6 +/- 15.1 pM* at 20 min). Portal PGE2 concentration increased gradually after the induction of acute pancreatitis, and differences from the control group were significant at 20, 30, and 180 min (1426 +/- 175 pM at 0 time, 1956 +/- 273 pM* at 20 min, 2148 +/- 265 pM** at 30 min, and 3369 +/- 686 pM* at 180 min). Portal histamine and serotonin concentrations increased somewhat, but not significantly. These findings suggest that the injection of bile into the pancreatic duct causes the pancreas to quickly release a large amount of bradykinin into the portal vein, which immediately reduces the pancreatic blood flow in the early phase, thus accelerating the progress of acute pancreatitis.

The fluidity of liver plasma membranes was assessed during regeneration following partial hepatectomy in rats. Fluorescence polarization was measured using 1,6-diphenyl-1,3,5-hexatriene to evaluate the fluidity of liver plasma membranes. The fluorescence polarization value of liver plasma membranes of control rats was 0.189 +/- 0.005 (mean +/- SEM). The fluorescence polarization values decreased initially after a 70% hepatectomy [0.176 +/- 0.003 at 3 hr and 0.174 +/- 0.004 at 6 hr (P < 0.01)] at 3 and 6 hours, but then recovered to preoperative levels (0.181 +/- 0.004, 0.189 +/- 0.004, 0.189 +/- 0.003, 0.193 +/- 0.003, 0.200 +/- 0.002, 0.208 +/- 0.007, and 0.190 +/- 0.003 at 12, 24, 36, 48, 72, 96 hr, and 7 days, respectively). Flow cytometric analysis using anti-bromodeoxyuridine (BrdU) monoclonal antibody was used to investigate the cell kinetics of the regenerating liver after 70% hepatectomy. The proportion of S-phase incorporating BrdU was not increased at 6 or 12 hr, but it then increased to peak of 27.9 +/- 1.9% at 24 hr. Subsequently, at 48, 72, and 168 hr, the proportion decreased to 8.5 +/- 0.6, 5.9 +/- 0.3, and 1.0 +/- 0.1%, respectively. The ratio of phospholipid to cholesterol contents of liver plasma membranes following partial hepatectomy did not significantly change in spite of markedly altered membrane fluidity. Thus, the alteration of membrane function occurs in conjunction with the initiation of the regeneration process following partial hepatectomy. It is suggested that the alteration of membrane fluidity plays an important role in the homeostatic response for cell proliferation.

The present study was designed to investigate the role of Kupffer cells (Kc) in the surveillance of liver tumors. We examined the antitumor activity of Kc by 51Cr releasing assay and inhibition of [3H]thymidine ([3H]TdR) incorporation into tumor cells. We also studied the change in the growth of liver tumors following the activation and the blockade of Kc. The cytotoxicity of Kc against K562 increased as the effector:target (E:T) ratio rose and reached its maximum level of about 18% at an E:T ratio of 20:1. [3H]TdR incorporation into target cells (P815 and AH130) was also inhibited by Kc. Such antitumor activity of Kc was augmented by OK432 (K562, from 13.8 +/- 5.6 to 21.9 +/- 2.5%; AH130, from 19.2 +/- 14.5 to 37.1 +/- 12.6%). In the experiment of the inoculation of AH130 via the portal vein, OK432 decreased the number of hepatic foci, whereas macrophage inhibitors carrageenan and gadolinium increased the number of tumor nodules. In addition, gadolinium injection reduced the number of Kupffer cells reactive with monoclonal antibodies directed against macrophages ED2 and Ki-M2R. Tumor growth in the liver was maximum in rats with both gadolinium treatment and splenectomy. In conclusion, Kc have antitumor activity, and augmentation of Kc may be a possible strategy to prevent hematogenous hepatic metastasis.

OBJECTIVE: This study compared the UICC classification with the General Rules for Gastric Cancer Study (GRGCS) of the Japanese Research Society by analyzing recent results of gastric cancer surgery in Japan. SUMMARY BACKGROUND DATA: The present UICC stage classification for gastric cancer was published in 1987 and the Japanese GRGCS were published in 1985. Both are based on the results of surveys conducted in the early 1970s. METHODS: The survival curves of 926 patients, who underwent gastric cancer surgery between 1982 and 1985 at Kyoto University Hospital and its 31 associated hospitals, were analyzed according to the UICC classification and the GRGCS using SAS computer software. RESULTS: There was no difference in survival rate between UICC stages IA and IB. GRGCS stage III was found to include UICC stages II, IIIA, and IIIB, and GRGCS stage IV included UICC stages IIIA, IIIB, and IV, with significantly different survival rates. In contrast, each UICC stage included different GRGCS stages with no significant differences in survival rates. The survival rate of stage IV patients of both classifications who underwent gastrectomy was significantly higher than that of stage IV patients receiving bypass or exploratory surgeries. CONCLUSIONS: The UICC classification is better than the GRGCS for classifying gastric cancer in Japan. However, UICC stage I does not need to be subdivided into stages IA and IB, and stage IV should be further subdivided into stages IVA and IVB according to the surgery performed: IVA, gastrectomy, and IVB, bypass or exploratory surgery.

This phase II study was designed to assess the therapeutic potential of intensive course treatment with three anticancer agents: 50 mg of cisplatin on day 1, 40 mg/m2 of epirubicin on day 2, and 250 mg of 5-fluorouracil on days 2 through 5. Drug courses were repeated every 2 weeks and most patients received between 4 and 6 courses. Thirty-five patients with measurable advanced solid cancers entered the study. They consisted of 16 gastric, 5 colorectal, 4 gallbladder, 3 pancreatic, 3 lung, 2 esophageal, 1 uterine, and 1 ovarian cancers. Of the 35 patients, 29 were evaluated for therapeutic effect of the regimen, and the overall response rate was 31.0% (5 CR + 4 PR/29). A 33.3% rate of tumor regression, consisting of 2 complete responses (CR) and 3 partial responses (PR) out of 15 patients (2 CR + 3 PR/15), was seen for gastric cancers. For the other types of tumors the responses were achieved in 2 lung cancers (1 CR + 1 PR/3). 1 uterine cancer (1 CR/1), and 1 ovarian cancer (1 CR/1). The esophageal, colorectal, pancreatic, and gallbladder cancers were unresponsive to this regimen. Toxicities of the drug treatment were clinically tolerable and consisted of general malaise, nausea, vomiting, stomatitis, alopecia, and leucopenia. However, two patients died of uncontrollable metabolic acidosis after 1 and 2 courses, respectively. This intensive course treatment appears to promote the regression of gastric, lung, and gynecologic cancers.

We evaluated the protective effect and the mechanism of action of the trypsin inhibitor, urinastatin, extracted from human urine, in experimental acute pancreatitis induced by a supramaximal dose of cerulein (5 micrograms/kg/hr for 3.5 hr). Urinastatin in a dose of 10,000 units/kg/hr was given by three different methods of continuous infusion: (1) 2 hr before and during cerulein infusion, (2) only during cerulein infusion, and (3) starting 1 hr after the beginning of cerulein infusion and continued for 3.5 hr. In protocol 1 and 2 urinastatin was significantly more protective than in 3. In protocol 1 urinastatin was very protective in all parameters tested (serum amylase level, pancreatic water and amylase content, distribution of lysosomal enzymes, cellular and lysosomal fragility). These results suggest that the administration of urinastatin before and during cerulein infusion may suppress the pathogenesis and evolution of pancreatitis by inhibiting the chain reaction of pancreatic enzyme activation closely related to redistribution of lysosomal enzyme and lysosomal fragility.

Islet transplantation with a bioartificial pancreas is a potential alternative to whole pancreas transplantation. The authors constructed a bioartificial pancreas using mesh reinforced polyvinyl alcohol hydrogel tubes (MRPT), in an attempt to clarify the in vitro responsiveness to glucose of islets seeded in the MRPT. When the MRPT were perfused in a small chamber with buffer containing 3.3 mmol or 16.7 mmol glucose, insulin release from the MRPT began to increase at 9 +/- 3 min, reaching a plateau at approximately 40 min after the glucose concentration in the perfusate increased from 3.3 to 16.7 mmol. When MRPT seeded with islets were subjected to static incubation in buffer containing 3.3 mmol or 16.7 mmol glucose, insulin release from the MRPT remained elevated for 3 hr of high glucose stimulation, the amount of secreted insulin depending upon the number of islets seeded. Although pre incubation of semipermeable membranes in culture medium containing fetal bovine serum prior to seeding with islets has recently been reported to improve insulin release, the authors found that such pre treatment of the MRPT did not have a beneficial effect. Their in vitro findings in this study suggest that the bioartificial pancreas using MRPT could be a promising therapeutic approach to human diabetes mellitus.

In patients with Stage II or III breast cancer and in patients with liver metastases from breast cancer, we examined cellular interaction in the cytotoxicity against autologous tumor cells by interleukin-2(IL-2)-cultured lymphocytes (CL) and fresh peripheral blood lymphocytes (FPBL) treated with immunochemotherapy including OK-432 and cyclophosphamide. In flow cytometric analysis, CD8 + CD11b+ and CD16+ cells significantly decreased after immuno-chemotherapy in both groups of patients. A protocol study in Stage II or III breast cancer patients showed suppressive activity of FPBL on the cytotoxic activity of CL in 3/9 of the non-treatment group but no suppressive activity and enhancing activity in 3/7 in the immuno-chemotherapy group. Moreover, in 19 patients with liver metastases from breast cancer treated with immuno-chemotherapy including adoptive immunotherapy, FPBL in 6/19 showed enhancing activity, and in 8/19 suppressive activity in the lysis of autologous tumor cells. In assaysin vitro using autologous and allogeneic tumor cells, FPBL showed a partial specificity in cellular interaction against autologous tumor cells. CD4-depleted FPBL inhibited cytotoxicity of CL, while CD8-depleted FPBL enhanced cytotoxicity of CL in patients with liver metastases. These results suggest that immuno-chemotherapy eliminates the suppressive population in FPBL and may induce tumor regression if combined with adoptive immunotherapy using CL.

Twenty-four patients with liver metastases from gastric or colorectal cancer were treated with OK-432-combined adoptive immunotherapy (AIT). Lymphocytes isolated from regional lymph nodes or peripheral blood were cultured with medium containing T cell growth factor and sonicated tumor extract antigen (SE-Ag) for 9–13 days. The cultured lymphocytes were transferred mainly through the hepatic artery after the administration of OK-432, a streptococcal preparation. Sixteen of the 24 patients received a low dose of anti-cancer agents between the OK-432 injection and cell transfer. When cultured without SE-Ag, regional lymph node lymphocytes (RLNL) showed significantly (P

We analysed superoxide anion (O2-) release from primary-cultured human hepatic macrophages (HHM phi) and peripheral blood monocytes (MO) derived from patients with normal and cirrhotic livers. Primary cultured human hepatic macrophages and MO from cirrhotic patients released less O2- than cells from normal patients. Superoxide anion release by HHM phi showed a significant, positive correlation with the serum glutamate pyruvate transaminase (GPT) level, whereas O2- release by MO showed only a weak correlation with the GPT level. In conclusion, a significant reduction in O2- release was observed in HHM phi and MO cultured from cirrhotic patients. Primary-cultured human hepatic macrophages are probably more susceptible to environmental changes within the cirrhotic liver than MO, since they are found locally within the liver and correlate with serum GPT levels.

This in vivo and in vitro study demonstrates the protective effects of a new synthetic protease inhibitornafamostat mesilate, FUT-175—on increased cellular and lysosomal fragility within acinar cells during the early stage of cerulein-induced acute pancreatitis in rats. FUT-175 prevented hyperamylasemia, pancreatic edema, congestion owing to amylase, and lactic dehydrogenase (LDH) discharge from acini as well as cathepsin-B leakage from lysosomes dose-dependently in doses of 1-10 mg/kg h. These results suggest that FUT-175 can protect against pancreatitis at subcellular levels in lysosomes and cellular or organelle membranes. Proteases may well play the important role in the pathogenesis of acute pancreatitis, and such a low molecular protease inhibitor may be useful clinically in the treatment of acute pancreatitis.

The effect of three anticancer agents, cisplatin, doxorubicin, and mitomycin, on liver regeneration after 70% partial hepatectomy in rats was investigated by total DNA content of the liver and flow cytometric analysis of hepatocyte nuclei using two-color staining of anti-bromodeoxyuridine monoclonal antibody and propidium iodide. Total DNA content of regenerating liver 7 days after hepatectomy showed significant suppression of regeneration by these agents (P less than 0.01). The inhibitory effects of the agents on the cell cycle of hepatocytes by flow cytometric analysis were (1) a delay of the peak or a decrease in the proportion of S phase nuclei and/or (2) polyploidization of the nuclei, demonstrated by accumulation of 8c and occasionally 16c nuclei, of which the DNA contents were four and eight times as much as that of diploid (2c) cell nuclei, respectively. The former (1) suggests G0 or G1 phase block, and the latter (2) G2 phase block. In terms of total DNA content of regenerating liver, the inhibitory effect was most prominent in the cisplatin-administered groups. The polyploidization of nuclei was most remarkable in the mitomycin-administered groups. Although the total DNA content recovered to the level of control at 6 weeks after hepatectomy, the polyploidization effect persisted in the drug-administered groups. These agents had no cytocidal action on proliferating hepatocytes as can be seen from the aspartate aminotransferase and alanine aminotransferase serum levels. We conclude that in a short-term observation, the anti-cancer agents significantly inhibit liver regeneration, although the inhibitory effect on DNA synthesis does not last long.(ABSTRACT TRUNCATED AT 250 WORDS)

The redistribution of cathepsin B, a lysosomal enzyme, from the lysosomal pellet to the zymogen pellet in the subcellular fractionation, the colocalization of cathepsin B with digestive enzyme, and increased cellular, lysosomal, and mitochondrial fragility within acinar cells have been found during the early stages of caerulein-induced acute pancreatitis in rats. In the present study, the authors investigated the protective effects of prostaglandin E1 and E2, a combined therapy of these prostaglandins, and a new, synthetic, low molecular weight protease inhibitor, ONO3307, on the exocrine pancreas in this noninvasive model of experimental pancreatitis in vivo and in vitro. Prostaglandin E2, but not E1, prevented hyperamylasemia, congestion of amylase and trypsinogen in the acinar cells, redistribution of cathepsin B, and amylase and lactate dehydrogenase discharge from the dispersed acini. It also prevented cathepsin B leakage from the lysosomes and malate dehydrogenase leakage from the mitochondria in an almost dose-dependent manner, particularly at the dose of 100 μ g/kg/hr continuous infusion. Furthermore, the combined therapy of prostaglandin E2 with ONO3307 strongly inhibited all the parameters tested in this study. This combination therapy seems to be the most effective against secretagogue-induced pancreatic injuries. These results indicate that cellular and subcellular organellar fragility seem to be closely involved in the pathogenesis of acute pancreatitis. Prostaglandin E2 seems to have important cytoprotective effects on the biologic membranes, such as a stabilizer of lysosomal or mitochondrial membranes. In addition, these findings also suggest the crucial roles of some unknown proteases in the etiology of acute pancreatitis, and indicate the clinical effectiveness of prostaglandins and this type of low molecular weight protease inhibitor for acute pancreatitis.

Through the analysis of accumulated experimental data of cell growth, a model of unrestricted cell growth is presented here. This model is based hypothetically on the cellular commitment to senescence as in the previously presented commitment theory. Cells are divided into 3 types, namely uncommitted cells, committed cells and terminal cells. The division of an uncommitted cell produces an uncommitted cell itself and a committed cell of the first generation. This committed cell goes through a limited number of cell divisions until the cells become non-dividing terminal cells. It is hypothesized that during a committed cell division, there is a very small probability that an uncommitted cell may be generated. With computer simulations, it is estimated that a committed cell of the first generation may divide around 30 times until they become non-dividing terminal cells, and that the probability of an uncommitted cell regeneration for each committed cell division may be 2 (−H) . This hypothesis may possibly clarify some aspects of the biological phenomena of cell growth, which can not be explained by the previous commitment theory, such as the growth pattern of cultured diploid cells, cancer initiation and promotion, and cancer progression and metastasis.

We developed a local AIT using PEL cultured with TCGF combined with preadministration of OK-432. Twenty-six patients of breast cancer with pleural effusion have been treated with this therapy since 1983. PEL expanded and tumor cells collapsed by day 9 in culture with TCGF. Cultured PEL possessed significantly higher cytotoxic activity against autologous tumor cells than PBL cultured in the same condition (p less than 0.05), but there was no difference between their cytotoxic activities against K562. The proliferation rate of PEL obtained after intrapleural administration of OK-432 was higher than that obtained before OK-432 (p less than 0.01). Moreover, the cytotoxic activities against both autologous tumor and K562 of cultured PEL obtained after OK-432 administration was significantly (p less than 0.05) higher than those cultured PEL obtained before. Cultured PEL (1 x 10(8)-6 x 10(9)) were transferred into the pleural cavity after the intrapleural administration of OK-432 (1-5 KE). The volume of pleural effusion increased temporarily after the administration of OK-432 but significantly (p less than 0.01) decreased after AIT. Tumor cells disappeared cytologically in 22 patients at the last puncture of pleural effusion. Pleural effusion disappeared completely in 19 of 26 patients and decreased by more than 50% in volume in 6 patients. Performance status improved in 22 patients. The response rate for OK-432-combined AIT in the present study was 96%. The survival period of the patients treated by OK-432-combined AIT in this trial was significantly (p less than 0.002) prolonged compared to that of the patients receiving chemotherapy alone.(ABSTRACT TRUNCATED AT 250 WORDS)

To remove regionally recurrent cancer of the rectum completely, more radical extensive resections were attempted than the conventional ones in 35 patients. Twenty-two patients underwent total pelvic exenteration. In 23, sacral resection was done, combined with other procedures including excision of the entire circumference of the lower pelvic ring. There were two hospital deaths. Delayed wound healing was seen in ten. Preoperative severe pain was alleviated in 18. Also, perineal ulcer, vaginal bleeding, bloody stools, and tenesmus disappeared. Of the four 5-year survivors (estimated rate, 23%), only one was disease-free. A second recurrence was seen in 27 patients, of whom 23 had regional failure. It was concluded that so-called local recurrence is a lesion more widespread than indicated by imaging and biopsy. Therefore, even the most radical resection rarely can cure this disease. However, such a surgical procedure can afford more effective palliation than other treatments and may prolong life in selected patients significantly.

Immunosuppressive effects of cyclosporin A (CsA) and FK 506 (FK) on peripheral blood lymphocytes were studied in dogs in respect to mixed lymphocyte reaction, proliferative responses to recombinant interleukin-2 (rIL-2), phytohemagglutinin (PHA) and concanavalin-A (Con-A); phenotypes of OKIa1, CD3, CD8 and surface IgM; cytotoxic activity against xenogeneic tumor cells. CsA (2.0 or 5.0 mg/kg, intravenously) or FK (0.16 mg/kg, intramuscularly) was given to mongrel dogs every morning for serial 21 days. The blood concentrations of CsA, measured as trough levels by fluorescence polarization method, ranged from 37 to 350 ng/ml in dogs administered at 2.0 mg/kg and from 170 to 894 ng/ml in dogs administered at 5.0 mg/kg during treatment, respectively. In dogs treated with FK at a dose of 0.16 mg/kg, the drug concentrations in the plasma during treatment ranged from 0.16 to 1.8 ng/ml. Mixed lymphocyte reaction and proliferative responses to rIL-2, PHA and Con-A, which were declined by CsA, were not affected by FK. In contrast, the proportion of OKIa1+ cells was not affected by CsA, whereas FK decreased the proportion of OKIa1+ cells progressively during the course of treatment. Cytotoxic activity was suppressed by both CsA and FK. These results possibly indicate that CsA and FK exert their immunosuppressive effects via different mechanisms.

Although disorders in bone metabolism have long been recognized as typical sequelae of gastrectomy, there has until now been no reliable method of providing precise assessment of bone mass, resulting in a variation of reported incidence. In this study, metabolic bone disease was evaluated in a well characterized population: 34 men 2–5 years after gastrectomy; 11 men 6–10 years after gastrectomy; eight men 2–5 years after colonic resection; and 115 healthy men. The age range was 50–69 years. An innovative reliable method of dual energy X-ray absorptiometry was used which allows quantitative assessment of bone mineral content. Measurement of lumbar spine bone mineral density revealed that the mean(s.e.m.) bone mineral density in patients who had had a gastrectomy (2–5 years 0.84(0.03) g/cm2, 6–10 years 0.85(0.05) g/cm2) was significantly lower than that of patients who had undergone colonic resection (0.96(0.04) g/cm2) or healthy men (0.96(0.03) g/cm2). This study demonstrates that a high incidence of bone atrophy is induced at the relatively early period of 2–5 years after gastrectomy.

The effect of biological response modifiers, such as interleukin-2 (IL-2) and streptococcal preparation OK432, on the functions of hepatic macrophages was investigated. The macrophages, even with no exogenous stimulation, produced superoxide anion (O2-) and tumor necrosis factor (TNF), displayed cytotoxicity against K562 cells and cytostasis against P815 cells and expressed immune-region-associated antigen (Ia). IL-2 administered in vitro or in vivo enhanced O2- production by hepatic macrophages and the intravenous injection of OK432 also enhanced O2- production. Furthermore, IL-2 added to the culture medium of hepatic macrophages isolated from OK432-injected rats augmented O2- production even more. The TNF production and Ia expression of the macrophages were also increased by the intravenous injection of OK432. As with O2- production, the cytotoxicity of the cells was enhanced by OK432 injection or by IL-2 added to the culture medium and the combination of OK432 and IL-2 augmented their cytotoxicity even more. Thus, the present study suggested that IL-2 and OK432 induce the augmentation of the antitumor activity of hepatic macrophages, partly as a result of the increase in production of O2- and TNF and Ia expression.

To establish useful predictors of the intrahepatic recurrence of hepatocellular carcinoma (HCC) after partial hepatectomy, retrospective analyses of clinical and pathologic factors were done in 112 of 206 patients treated by partial hepatectomy. The absence or presence of intrahepatic recurrence was confirmed by a follow-up study. Cancer-free survival rates after 1, 2, 3, and 5 years were 54.8%, 36.7%, 32.5%, and 25.6%, respectively. The significant factors affecting recurrence were tumor size, number of tumors, cancer cell infiltration of the fibrous capsule of the tumor, portal involvement, and stage of the tumor, but the grade of anaplasia according to Edmondson-Steiner's classification and the severity of associated liver cirrhosis did not show a correlation with the incidence of recurrence. According to Akaike's Information Criteria (AIC), tumor number is useful for predicting early prognosis, and capsular infiltration is a good indicator of long-term survival. However, portal involvement gives much prognostic information throughout the entire postoperative period. Cancer 1992; 69:913–919.

The response and survival of 26 patients with liver metastases from breast cancer, who received OK-432-combined adoptive immunotherapy from 1984 to 1990, were evaluated. OK-432-combined adoptive immunotherapy was comprised sequential treatment via the hepatic artery with a streptococcal preparation, OK-432 (1-5 KE), and adoptive transfer of lymphocytes expanded in T-cell growth factor and sonicated tumor extract antigen. Seventeen (65%) patients responded to the therapy. The median survival time of all patients after treatment was 13 months (range, 2-63 months). Of the 20 prognostic factors analyzed, performance status (PS) alone was related to response (P less than 0.01). The response rate of the patients with a PS of 0-2 was 83% but only 25% in those with a PS of 3 or 4. In univariate analysis, 11 factors significantly influenced the survival: tumor response; size of primary tumor; menopausal status; PS; serum bilirubin, albumin, lactate dehydrogenase and glutamate-oxalate transaminase (aspartate aminotransferase); the extent of liver involvement; and the number and the proliferation rate of transferred lymphocytes. The MST was 22.8 months for the responders versus 2.8 months for the nonresponders (P less than 0.01). In multivariate analysis, the most important factor associated with survival was the tumor response, as well as PS, liver involvement, lactate dehydrogenase and albumin. These results suggest that OK-432-combined adoptive immunotherapy can be considered a candidate for a randomised control study and these factors should be used for stratification.

To define the role of free radicals and of lipid peroxide involvement during the progress of cerulein-induced acute pancreatitis in mice, we evaluated the effect of a novel free radical scavenger, 2-octadecylascorbic acid (CV-3611), on pancreatic edema formation, and the levels of serum enzymes (amylase, lipase) and of lipid peroxide in pancreatic tissue. Mice were divided into three groups: control group, intraperitoneal injection of saline only; pancreatitis group, cerulein 50 micrograms/kg injected intraperitoneally six times at 1-hr intervals; treatment groups, CV-3611 10 mg/kg subcutaneously just after intraperitoneal cerulein injection. After the cerulein injection, the degree of pancreatic edema formation, serum amylase and lipase levels, and the amount of lipid peroxide in pancreatic tissue increased significantly during the observation period of 12 hr. Treatment with CV-3611 resulted in significant reduction in pancreatic edema formation at 3.5 hr (P less than 0.05) and 9 hr (P less than 0.05), serum amylase and lipase levels at 3.5 hr (P less than 0.05) and 12 hr (P less than 0.05), and lipid peroxide levels at 3.5 hr (P less than 0.05), 6 hr (P less than 0.05) and 12 hr (P less than 0.05). These results indicate that a novel free radical scavenger, CV-3611, has a strong therapeutic effect during the development of acute pancreatitis and suggest that oxygen-derived free radicals play an important role in the pathogenesis of acute pancreatitis.

Twenty-one esophageal cancer cell lines (KYSE series) have been established from the resected specimens of patients with esophageal cancer. Three lines, KYSE-30, KYSE-50, and KYSE-70, were derived from the implanted tumor of nude mice (initial passage); others were derived from resected specimens. Each cell line was morphologically distinct. Detailed cytogenetic analysis indicated that each cell line was karyotypically unique, and DNA fingerprint analysis showed no cross-contamination among cells. Doubling time ranged from 13.7 to 75.5 hours, and modal chromosome numbers ranged from 46 to 120. Most cell lines grew in monolayer, but two cell lines (KYSE-50 and KYSE-360) grew as floating cell aggregates. No correlation was demonstrated between the establishment of cell lines and cell differentiation. These cell lines are the first reported to be homogeneous and individually unique and may provide a useful model for the study of human esophageal cancer.

In order to learn more about how human hepatic macrophages function, we analyzed the effect of exogeneous PGE2 on the amounts of interleukin-1 (IL-1) and superoxide (O2−) released from primary-cultured human hepatic macrophages (HHMΦ). When endogenous PGE2 production was blocked by indomethacin, exogenous PGE2 reduced IL-1 release from HHMΦ in a dose-dependent manner, whereas it tended to increase O2- release from HHM Φ. These results may suggest the probable contribution of PGE2 in regulating HHMΦ mediator release in vivo.

胸部食道癌切除術前の胃と術後再建胃管の胃底部と前庭部の2点で24時間pHモニタリングを行った.全測定期間中pH頻度分布曲線を, 1型 (高酸型), 2型 (中間型), 3型 (低酸型), 4型 (前庭部高酸型) に分類し, 各型群間で胃内酸度を術前後で比較検討した.深夜, 食事中, 食後の各期間pH中央値は各群間で特徴を有し, この分類法が酸分泌の解析上有用と考えられた.深夜胃底部pHは1, 2型群で低値となり, 1, 2型群において, 深夜胃底部pH中央値の平均, 深夜胃底部pHが3以下となる時間の割合には術前後で有意差はなかった.術後長期経過例でも深夜胃底部は低pHを示す症例が多く, 再建胃管の夜間酸分泌が保たれており消化性潰瘍発生に注意すべきと考えられた.また, 術後1, 2型群深夜前庭部のpHが3以下となる時間の割合と空腹時血清gastrin値の間の有意な逆相関 (p

遠隔転移のみられない膵癌において, 治癒切除を行うためには術前の画像診断, とくにCT, 血管造影所見から膵後面浸潤, 血管浸潤の程度を適格に把握する必要がある.上腸間膜動脈などの主要動脈浸潤がなけれぼ切除可能と判断されるが, その際は門脈系浸潤の有無を問わず, 膵に接する門脈系は周囲神経, リンバ組織を含む結合織とともに一塊として切除する拡大切除を行う.とくに血管周囲郭清は大動脈周囲, 上腸間膜動脈周囲を含め徹底して行い, 併せて局所再発遠隔転移に対する予防策が必要である.

膵管結紮家兎にて膵腺房細胞の変化と新しい蛋白分解酵素阻害剤E3123の保護効果を検討した.3時間の短期間膵管閉塞とsecretin (0.2CU/kg・hr) 投与にて, 門脈血中amylase (46±4U/ml) (p

教室の過去10年間の手術施行上部胆道癌133例を対象として, 在宅生存率を, stage, 治癒度, 術式別に検討した.在宅生存率は, stageの進行と一致して低下し, stage IVの1年在宅生存率は, 胆嚢癌7%, 胆管癌4%と低かった.治癒度からは, 絶対非治癒群では, 胆管癌0%, 胆嚢癌4%と低いが, 切除により, 前者で30%前後と上昇, 後者では相対非治癒8%, 治癒切除群で65%前後に上昇し.StageIII, IVの進行胆道癌に対する術式別では, 胆管癌, 胆嚢癌の姑息, 縮小, 拡大手術群の1年在宅生存率は, それぞれ, 0%, 25%, 22%;4%, 42%, 22%であった.以上, 切除により, quaiity of life (QOL) の改善は認められるものの, 拡大手術は必ずしも, 現段階では, QOLの改善には結びつかず, 今後, 拡大手術の適応について検討すべきであると考えられた.

An en bloc resection of esophageal cancer is one of the most radical forms of esophagectomy, and includes the resection of the thoracic duct, but a relatively high hospital mortality rate has been reported. There is very little knowledge on the pathophysiological changes after resection of the thoracic duct. We examined 24 patients who underwent en bloc resection. Some patients developed severe tachycardia or shock postoperatively which subsided after a massive infusion of plasma. Analysis of the fluid balance revealed that much more fluid was necessary during surgery and the postoperative 24 h than in patients treated by a standard esophagectomy. Postoperative lymphangiography or CT revealed abnormal collateral lymphatics around the kidneys or in the pelvic cavity. This suggests the development of the lymphaticovenous shunts, which differed depending on the anatomy of each patient. One patient with chronic hepatitis developed uncontrollable ascites. These are important findings which can hopefully reduce the high rate of hospital death after this operation.

Two-color flow cytometric (FCM) analysis using anti-bromodeoxyuridine (BrdU) monoclonal antibody (MoAb) was used to investigate the cell kinetics of regenerating liver after 70% partial hepatectomy in rats. Three peaks were seen in DNA histograms of rat hepatocyte nuclei, corresponding to diploid(2c), tetraploid(4c), and octaploid(8c). These proportions changed in the course of regeneration which were clearly demonstrated by DNA histograms using flow cytometry. The proportion of diploid, tetraploid, and octaploid nuclei in control liver were 49.3 +/- 1.6%, 45.0 +/- 7.4%, and 1.7 +/- 0.7%, respectively. A significant change occurred at 24 hours after hepatectomy, as FCM revealed 25.9 +/- 1.1% diploid, 54.5 +/- 1.2% tetraploid, and 9.0 +/- 0.9% octaploid. This shift to polyploid nuclei persisted until 72 hours, and then gradually returned to the pattern of control liver. The S-phase nuclei which incorporated BrdU increased rapidly at 24 hours to a peak of 11.3 +/- 0.9%, and gradually decreased to 5.8 +/- 0.8%, 5.3 +/- 0.8%, 2.4 +/- 0.6%, 2.9 +/- 1.1%, and 1.2 +/- 0.6%, at 48, 72, 96, 120, and 168 hours, respectively. This 2-color FCM analysis made a detailed analysis of the cell kinetics in regenerating hepatocytes possible, and may be applied in investigations of various aspects of liver regeneration.

This study was designed to evaluate the acute toxic effects of cyclosporin A on the exocrine pancreas and the protective effects of a new potent protease inhibitor, 4-(2-succinimidoethylthio) phenyl 4-guanidinobenzoate methanesulfonate, E3123. Cyclosporin A in a dose of 5 mg/kg.h caused a significant increase in serum amylase levels, pancreatic amylase content, and interstitial edema, suggesting that it induces exocrine pancreatic injury. Cyclosporin A also caused redistribution of cathepsin B from the lysosomal fraction to the zymogen fraction, indicating colocalization of lysosomal enzymes with pancreatic digestive enzymes. E3123 in a dose of 2 mg/kg.h prevented almost completely these changes caused by cyclosporin A. These results indicate that exocrine pancreatic injury is induced by cyclosporin A and that lysosomal enzymes play important roles in the pathogenesis of this injury and also suggest that E3123 might protect the exocrine pancreas of patients receiving cyclosporin A after organ transplantation.

In order to decrease the perioperative complications by preoperative cisplatin chemotherapy, the preoperative single administration of cisplatin (30 mg/m2) was performed weekly from one to six times in 36 consecutive patients with esophageal cancer classified as higher than Stage II. The survival curve of 17 patients in Stage III was significantly better (P < 0.05) than that of patients who had been treated without preoperative cisplatin treatment. In 3 of the 12 patients who had locally invasive cancer, either the main tumors or the metastatic lymph nodes, which had invaded the trachea or the left main bronchus, sufficiently receded, so that a curative esophagectomy became possible; 2 of them have survived over 33 months while 1 died of pneumonia 33 months after surgery. The number of perioperative complications was minimal, and thus, we consider that the postoperative use of cisplatin and fluorouracil is indicated in patients in whom a histological response is noted in the resected specimens.

The effect of Mitomycin-C (MMC) and Adriamycin (ADM) on the antitumor-associatted function of Kupffer cells was examined. MMC and ADM enhanced the production of superoxide by Kupffer cells in cultures at low concentrations likely to occur in clinical use. The expression of interleukin-2 receptor, Ia antigen and asialoGM1 antigen, measured by flowcytometry, was increased by contact with MMC. Growth inhibition of AH130, rat ascites hepatoma, and P815, murine mastocytoma, by Kupffer cells treated with anticancer drugs was greater than that by Kupffer cells alone or anticancer agent alone. These results show that MMC and ADM activate Kupffer cells, leading to synergistic antitumor activity. The results suggest that some anticancer agents act through immunological mechanisms as well as through direct antineoplastic activity.

直腸孤立性潰瘍症候群に対し, 経仙骨的に広範な直腸粘膜環状抜去術を施行した症例を報告する.症例は20歳男性で, 8年前から持続する血便, 粘液便, 裏急後重を訴え, 内科治療や排便時のいきみを控えても軽快しなかった.下部直腸に多発する隆起性病変を認め, 生検にて粘膜固有層のfibromuscular obliterationの所見を得, 直腸孤立性潰瘍症候群と診断した.術式は経仙骨的に直腸後壁を切開後, 粘膜のみを環状に剥離後切除し, 露出した直腸筋層を縦に縫縮後, 粘膜断端を再吻合した.術後の直腸肛門内圧測定でも異常は認められず, 患者は術後25か月現在無症状である.本症例はこの術式で治癒した第2例目である.

The present study was designed to investigate the pathophysiology of obstructive jaundice by analyzing the function of hepatic macrophages and their role in immune responses and homeostasis in rats. The phagocytic index, determined by the rate of disappearance of 51Cr-endotoxin from the peripheral blood after intravenous injection, was increased in obstructive jaundice 2 weeks after bile duct ligation. The superoxide production of isolated hepatic macrophages and peripheral blood monocytes, measured by the superoxide dismutase inhibitable ferricytochrome c reduction method, was increased. Prostaglandin E2 release, measured by RIA, was markedly increased in rats with obstructive jaundice, but there was no significant difference in interleukin-1 release between jaundiced and control rats. The flow-cytometric analysis of surface molecules of hepatic macrophages showed decreased expression of interleukin-2 receptor in rats with obstructive jaundice. Thus, the functions of hepatic macrophages in rats with obstructive jaundice were impaired. This malfunction may disturb the immunoregulatory network and metabolism, although the exact implications of the altered function of hepatic macrophages have not yet been clarified.

Five human digestive organ cancer xenograft lines were subcutaneously transplanted in nude mice, and 7 anticancer agents (ACAs) were intravenously administered singly or in combinations at 4 times the standard clinical doses. Cisplatin (P) was effective in 4 lines and 5-fluorouracil (F) was effective in 3 lines. A total of 8 combinations with 2 effective ACAs were assessed in 4 lines and only 1 combination (FP) resulted in an augmented antitumor effect in a single line. A total of 11 combinations with 1 effective ACA plus 1 ineffective ACA were assessed in 2 lines, and only 1 combination (F + carboquone [CQ; Esquinon]) resulted in an augmented effect in a single line. A total of 5 combinations with independently ineffective ACAs were assessed in 1 line, and 1 combination (FP) resulted in augmented antitumor activity. No 2-ACA combinations showed enhanced side effects. Two combinations with 3 effective ACAs were assessed in 2 lines and 1 combination (FP + mitomycin C) resulted in an augmented effect with a greater mortality rate. A total of 12 combinations with both effective plus ineffective ACAs were assessed in 2 lines, and 3 combinations (FP + Adriamycin or FP + CQ or FP + etoposide) resulted in an augmented effect in 1 line with enhanced side effects. These results indicate that the combination of independently effective ACAs does not always cause an augmented antitumor effect, and may sometimes result in augmented side effects. According to this study, the 3-ACA combinations with FP + 1 effective agent appear to be the best regimens for digestive organ cancer.

We investigated the production of chemical mediators by hepatic macrophages from rats with sepsis and the modulation of hepatocyte function by these hepatic macrophages. The chemical mediators we measured were superoxide (O 2 − ), TNF, IL-1, and PGE2. Production of these mediators by hepatic macrophages from rats with sepsis was significantly increased. Furthermore, protein synthesis by cultured hepatocytes was inhibited in a co-culture system of hepatocytes and hepatic macrophages from rats with sepsis, and it was even inhibited by the supernatant of cultured hepatic macrophages from septic rats. These results demonstrate that hepatic macrophages are activated in sepsis and may play a role in inducing hepatic dysfunction in sepsis.

The present study was designed to evaluate the effect of rTNF alone or in combination with other BRMs on human digestive organ cancers. Six kinds of human digestive organ cancer xenografts (esophageal, stomach, colonic, pancreatic, bile duct, and liver cancers: EC-YO, GC-YN, CC-KK, PC-HN, BDC-SN and Li-7, respectively) were transplanted in nude mice, and rTNF was administered at 10(3), 5 x 10(3), or 10(4)U/head directly into the tumor 3 times a week for 2 weeks. EC-YO was the most sensitive to rTNF, and intratumoral administration of rTNF at 10(3) U/head caused tumor regression. PC-HN, CC-KK and GC-YN were relatively sensitive to rTNF, and their growth was significantly inhibited by rTNF at 5 x 10(3) U/head, however, the tumors regrew after treatment. Li-7 and BDC-SN were resistant to rTNF. The effects of rTNF in combination with recombinant interferon-gamma (rIFN-gamma), recombinant interleukin-2 (rIL-2), or streptococcal preparation OK-432 were assessed in mice transplanted with GC-YN. All combinations of rTNF at 5 x 10(3) U/head and other BRMs were more effective than rTNF alone, and GC-YN tumors were completely regressed after treatment with a combination of rTNF and rIFN-gamma or rTNF and OK-432. However in all cases, the combination of rTNF at 10(3) U/head and any other BRM did not improve the effect. Furthermore, the adverse effects of the combinations were more serious than those of rTNF alone. TNF may still be a useful cytokine, because it can induce the regression of tumors. However, for its clinical application, a method should be developed to reduce its side effects.

A murine plasmacytoma MOPC 104E (MOPC) is highly sensitive to chemotherapeutic agents such as cyclophosphamide and mitomycin C as well as to immunotherapy (OK-432-combined adoptive immunotherapy using interleukin-2-cultured killer cells). In the present study, we prepared cyclophosphamide-resistant MOPC cells (MOPC-CPA/R) by serial in vivo passage of tumor cells following cyclophosphamide treatment. The in vivo sensitivity of MOPC-CPA/R to mitomycin C or to immunotherapy (OK-432-combined adoptive immunotherapy) was significantly decreased compared to the parent MOPC. In vitro experiments showed that MOPC-CPA/R were more resistant (five-fold) to lysis by cultured immune spleen cells than MOPC. Inhibition of the lytic activity of cultured immune spleen cells against MOPC was significantly increased (P less than 0.05) by the addition of unlabeled MOPC compared to unlabeled MOPC-CPA/R. These results suggest that MOPC-CPA/R express weaker antigenicity than MOPC. However, the transfer of immune spleen cells cultured with tumor extract derived from MOPC-CPA/R significantly prolonged the survival of MOPC-CPA/R-inoculated mice. Thus, by repeated cyclophosphamide treatment, tumor cells with low-antigenicity were selected. These tumor cells had lower sensitivity to another chemotherapeutic agent and immunotherapy. Such an immunological response may play an important role in cancer therapy.

Effects of synthetic human pancreastatin-52 and human pancreastatin-29 on pancreatic secretion and blood flow were examined in rats and dogs. Synthetic human pancreastatin-52 and human pancreastatin-29 were equally potent in suppressing the release of amylase stimulated by cholecystokinin in rats in vivo. However, neither human pancreastatin-52 nor human pancreastatin-29 altered basal and cholecystokinin-stimulated amylase release from isolated dispersed rat pancreatic acini. In studies in dogs, human pancreastatin-29 suppressed releases of amylase and protein stimulated by cholecystokinin, but did not alter pancreatic blood flow. These results suggest that the inhibitory effects of pancreastatin on pancreatic secretion do not involve a direct action on pancreatic acinar cells nor alteration of pancreatic blood flow. Pancreastatin probably is important in regulating exocrine pancreatic secretions as well as endocrine pancreatic secretions.