[Age-related changes and sex differences on the serum chemistry values in Sprague-Dawley rats--I. 6-30 weeks of age].

Age-related changes of 27 items in serum chemistry were investigated in Sprague-Dawley rats of both sexes from 6 to 30 weeks of age. The following 12 items were shown as an increase in those values during growth and maturity, i.e., total protein, albumin (female only), glucose, total cholesterol, triglyceride, phospholipid (female only), beta-lipoprotein, cholinesterase (female only), asparate aminotransferase (female only), creatinine, direct-bilirubin and total-bilirubin. However, the following 4 items decreased with aging, i.e., asparate aminotransferase, alkaline phosphatase, creatine phosphokinase and inorganic phosphorus. No age-related changes were found in the values for calcium, sodium and chloride in both sexes and for alanine aminotransferase, cholinesterase and albumin in males. The sex differences were shown in the following 12 items: higher values in males were alkaline phosphatase, creatinephosphokinase, glucose and inorganic phosphorus, and higher values in females were cholinesterase, albumin, phospholipid, non-esterified fatty acid, urea nitrogen, direct-bilirubin, total-bilirubin and serum iron. No sex-related differences were found in the values for calcium, sodium, chloride and total cholesterol.

Spectral analysis of body weight, food and water consumption and spontaneous motor activity in male Sprague-Dawley rats.

Spectral analyses by maximum entropy method were performed on the time-series data of body weight, food and water consumption and spontaneous motor activity in Sprague-Dawley rats. These parameters were recorded by a monitoring system collected every 30 min for 10 days. All the parameters had circadian (24 +/- 4 hr) and ultradian (less than 20 hr) rhythms. From the logarithmic transformation of spectral frequencies and their power spectral densities, the characteristics of the spectrum of body weight showed 1/f fluctuation and those of food and water consumption were white noises. The spectrum of the spontaneous motor activity showed complex characters. The range of frequencies lower than 0.1 Hz showed white noise while the range of those higher than 0.1 Hz were 1/f or 1/f2 fluctuations, especially in the range between 0.5 and 1.0 Hz (1-2 hours cycle). The auto-correlation coefficients of body weight did not declined suggesting that 24 hours cycle is only periodic phenomenon in body weight changes. Though the auto-correlation coefficients of food and water consumption also did not declined, they had not enough auto-correlations. This was consistent with the appearance of white noise in spectral analyses. The auto-correlation coefficients of spontaneous motor activity declined remarkably, and this suggests that there are various periodic changes in spontaneous motor activity including one to two hours cycles. These results suggest that the possibility of further extension of research is expected by using rhythmicity. It is also indicated that the time of measurement of body weight must be taken into consideration in experiments because of the presence of the circadian rhythmic changes.

Diurnal variations in blood chemical items in Sprague-Dawley rats.

Diurnal variations in 23 blood chemical items were investigated in Sprague-Dawley rats of both sexes at 6 weeks of age. Blood samples were obtained every two hours. One to three periodic components detected by the maximum entropy method from the observed time-series and the optimum fitting parameters were calculated by the least squares method. The fitting curves were compared with the observed data for the evaluation of systematic and fluctuating parts of the time domain as circadian variations. The periodic components were detected from all the blood chemical items examined. This suggests that all items have systematic parts as basic rhythmic changes. The optimum fitting curves corresponded to the observed time-series in the following items: glucose, total protein, A/G ratio, alkaline phosphatase, Na, K, Cl and BUN of both sexes and total cholesterol, triglyceride, cholinesterase, CPK, inorganic phosphorus, direct and total bilirubin, creatinine and uric acid in males. Others showed insufficient correspondence to many fluctuating parts. This randomness was more frequently seen in females than in males.

Correlation of body weight gain with food and water consumption and spontaneous activity in rats.

We developed a data acquisition system which measure the body weight, food and water consumption and spontaneous activity of a rat automatically. The correlation of body weight gain with other parameters obtained by this system were analysed. Each parameter showed rhythmic circadian changes. The multiple regression analysis with stepwise method brought the high coefficiency on the body weight gain with water consumption in the dark period, whereas only low correlation was observed among the body weight gain, food consumption and spontaneous activity in the light period. Since the body weight is affected by various factors, food and water consumption, spontaneous activity and/or other parameter(s) with body weight should be monitored simultaneously.

[Combined effect of glucocorticoid and TJ-114 (Tsumura Sairei-to)].

TJ-114 (Tsumura Sairei-to) is a powdered extract made from 12 Chinese herbal drugs. TJ-114 is used against various nephrotic diseases and used as an inhibitory treatment for the side effects of glucocorticoids. We expected that TJ-114 would increase the survival ratio of rats given a high dose of glucocorticoid. Therefore, in this study, we investigated the combined effects of glucocorticoid and TJ-114 in rats. An ointment containing fluocinolone acetonide (FA) was applied on the back of 7-week-old Wistar male rats for various periods. TJ-114 was administered orally at the doses of 0.5, 1.0 or 2.0 g/kg simultaneously. The combination with TJ-114 suppressed the loss of body weight by FA. The survival ratio of the FA group was 50%, but it was 83% for the group treated with FA in combination with 0.5 g/kg TJ-114 and no deaths were observed in the other drug combined group. With a lower dose of FA, we investigated its hematological effects and determined the white blood cell (WBC) count. Although the lymphocytes were decreased by FA, the combination with TJ-114 depressed this decrease of lymphocytes significantly. Furthermore, TJ-114 significantly suppressed the insulin increase elicited by FA. Macroscopic observations showed atrophy and decreases in the weights of the thymus, spleen and adrenal, all being target organs of glucocorticoid. The combination with TJ-114 decreased these effects of FA. Moreover, microscopic examinations revealed that FA induced the degeneration of lymphocytes and lymphocyte depletion in the cortex of the thymus, caused atrophy of the white pulp, decreased the extramedullary hematopoiesis of the spleen, and caused the atrophy of zona fasciculata in the adrenal cortex. The combination with TJ-114 depressed these effects significantly. These results suggest that TJ-114 suppresses the adverse side effects of gucocorticoids.

Predictive factors for intrahepatic recurrence of hepatocellular carcinoma after partial hepatectomy.

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.

Automatic monitoring system for the measurement of body weight, food and water consumption and spontaneous activity of a mouse.

The simultaneous recording system for body weight, food and water consumption and behavior (spontaneous motor activity and drinking and feeding behavior) of a mouse was developed. The body weight and food consumption were measured by force transducers. Food and water consumption and drinking and feeding behavior were measured by an infrared luminous diode and a phototransistor. Spontaneous motor activity was measured by photosensors. The system control and data acquisition were performed by using a personal computer. Every parameter could be monitored with a desired time interval. All the data collected by this system revealed apparent circadian rhythm. In conclusion, this system would be a powerful tool for pharmacological and/or toxicological research.

[Effects of Shosaiko-to-go-keishikashakuyaku-to (TJ-960) on the valproic acid induced anomalies of rat fetuses].

Shosaiko-to-go-keishikashakuyaku-to (TJ-960) is an extract of nine herbal drugs (Paeoniae radix, Cinna momi cortex, Bupleuri radix, Zingiberis rhizoma, Glycyrrhizae radix, Ginseng radix, Scutellariae radix, Pinelliae tuber and Zizyphi fructus) that has a potent anticonvulsant action. The rat fetuses treated orally with TJ-960 during the organogenesis period (days 7-17 of gestation) revealed no anomalies (up to 3000 mg/kg/day). When TJ-960 was co-administered with sodium valproate (VPA, 400 mg/kg) during the organogenesis period, embryonic resorption, fetal body weight, ossification and skeletal variation or anomalies induced by VPA were markedly reduced. The maternal plasma and embryonic concentration of VPA with TJ-960, however, were not significantly different from VPA alone. These results suggest that TJ-960 has protective effects against the teratogenicity of VPA.

Clinicopathological features of malignancy in hepatocellular carcinoma.

The clinicopathological features showing the malignancy of hepatocellular carcinoma were investigated by retrospectively analyzing the postoperative prognosis after hepatic resection. The long-term prognosis was strongly affected by the existence of portal tumor invasion or intrahepatic metastasis as indicated by the following results. The 3-year cumulative survival rates were 61% and 38% for patients in portal vein tumor invasion groups (Vp0 and Vp1 (P less than 0.05). No patients with Vp2 or Vp3 could survive beyond 3 years after hepatic resection. Similarly, patients with intrahepatic metastasis IM0 showed a better prognosis, compared to those with IM2 or IM3 (P less than 0.05). In addition, the grade of tumor cell anaplasia to some extent affected the prognosis, but not the tumor growth pattern at the tumor/non-tumor boundary. The tumor growth rate, estimated by the alpha-fetoprotein doubling time, was not connected with venous invasion or intrahepatic metastasis, but it became shorter at the time of a recurrence. It is concluded that, from the standpoint of a long-term prognosis, the pathological features showing malignancy appear in venous invasion and intrahepatic metastasis.

[Clinicopathological evaluation of angiograms in hepatocellular carcinoma].

Hepatocellular carcinoma (HCC) was classified into the three types of expansive type "Exp", mixed type "Mix" and infiltrative type "Inf" according to the preoperative angiographic characteristics, referred to clinicopathological features. "Exp" type had less incidence of accompanied portal venous invasion and daughter nodules in comparison to "Inf" type. And "Exp" type had a better prognosis in the cumulative survival rate than "Inf" type (p less than 0.001). Thus, the present typing essentially classified by tumor vascular architecture represents "malignancy grade" of HCC. In addition, "Inf" type may be recommended more extensive resection or multidisciplinary therapy (for example TAE) for better prognosis.

[Indications for limited hepatic resection in hepatocellular carcinoma].

Indications for limited hepatic resection in patients with hepatocellular carcinoma were evaluated by retrospective and clinicopathologic analysis of postoperative cases. Particular attention was paid to hepatic functional reserve and long-term prognosis. In our department, in order to estimate the hepatic functional reserve, indocyanine green clearance test (ICG), Child's classification and the oral glucose tolerance test have been applied. For instances, the incidences of hospital deaths were 17% and 19% in cases with an ICG-K value below 0.06 and between 0.06 and 0.08, which were high, compared with 6% and 3% in cases ICG-K values of with 0.08-0.10 and 0.10-0.12. On the other hand, by clinicopathologically analyzing 18 surviving patients or those surviving without recurrence for over 3 years after hepatic resection and 41 cases of small hepatocellular cancer less than 3 cm in diameter, those with capsule formation and without portal tumor invasion, intrahepatic metastasis and capsular invasion had a good long-team prognosis without recurrence even following limited resection. Particularly, patients with a surgical margin of over 1cm between the tumor and cut surface were candidates for long, non-recurrence survival. Form these results, the indications for limited hepatic resection for hepatocellular carcinoma were considered to be on ICG-K value of less than 0.08 in hepatic function and cases without venous invasion and extracapsular tumor invasion. In other cases, it is recommended that limited resection should be followed by multidisciplinary treatment.

Usefulness of selective arterial secretin injection test for localization of gastrinoma in the Zollinger-Ellison syndrome.

Secretin was injected into a feeding or nonfeeding artery of a gastrinoma and blood samples were taken from the hepatic vein (HV) or a peripheral artery (PA) to measure the changes of serum immunoreactive gastrin concentration (IRG). The IRG in the HV rose within 40 seconds and in the PA rose within 60 seconds after the injection of secretin into a feeding artery, but not after secretin was injected into a nonfeeder. These results indicated that secretin directly stimulates a gastrinoma to release gastrin in vivo. The selective arterial secretin injection test (SASI test) was applied in three patients in whom gastrinomas could not be located by computed tomography, ultrasonography, or arteriography, and functioning gastrinomas were located in all three patients. In one patient, malignant gastrinomas in the head of the pancreas and in the duodenum could be resected radically with the help of this test.

[Selective arterial secretin injection test for localization of gastrinoma].

We have already shown that gastrinomas release gastrin when stimulated with secretin in vitro. In order to ascertain whether this is true in vivo and whether the reaction is clinically useful in the localization of gastrinomas, secretin was injected into a feeding artery of the gastrinoma in four patients, and blood samples were taken from a peripheral artery (PA) and the hepatic vein (HV) for determination of immunoreactive gastrin (IRG) and immunoreactive insulin (IRI) levels. When secretin was injected into a feeding artery of the gastrinoma, IRG rose within 40 seconds in the HV and within 60 seconds in the PA. When secretin was injected into a nonfeeder, IRG did not rise for 2 min in the PA. This test was performed to two of the postgastrectomized patients and a patient with hypergastrinemia due to atrophic gastritis. In these patients, IRG in the hepatic vein did not rise for two minutes. It was concluded that secretin directly stimulates gastrinomas to release gastrin in vivo and that the selective arterial secretin injection test is helpful in determining the location of the gastrinoma.

Short-term changes in blood ketone body ratios in the phase immediately after hepatic artery embolization: their clinical significance.

Changes in arterial and hepatic venous blood ketone bodies were investigated following transcatheter hepatic artery embolization (THAE) in patients with hepatocellular carcinoma. Acetoacetate/ beta-hydroxybutyrate ratio (ketone body ratio) in arterial blood was positively correlated with those of hepatic venous blood (r = 0.960, p less than 0.001), which reflects the mitochondrial redox potential in the embolized lobe. Nine cirrhotic patients were classified into three groups according to the changes in arterial blood ketone body ratio following THAE: Type A without decrease to below 0.7; Type B with a transient decrease to below 0.7, followed by its restoration within 5 hours; and Type C with decrease to below 0.7 without recovery within 5 hours. There were no serious complications in Type A and B patients. By contrast, severe sepsis and hepatic failure developed in Type C patients, possibly due to the extended embolization of both lobes. It is suggested that THAE can be successfully performed even in severely cirrhotic patients, as long as the embolized area is restricted to one lobe. In addition, changes in arterial blood ketone body ratios can give early information about the likely consequences of the THAE procedure just performed.

Portal arteriovenous fistula following partial gastrectomy.

An emergency transthoracic transection of the esophagus was performed on a 62 year old man for hematemesis caused by rupture of esophageal varices. Hepatic functional reserve of this patient was classified as Child C. Postoperative celiac arteriography revealed an arteriovenous fistula between a branch of the gastroduodenal artery and a branch of the superior mesenteric vein. He had undergone a partial gastrectomy with Billroth II reconstruction 15 years earlier. Bleeding from the esophageal varices did not recur, but he subsequently died of liver failure. In some areas of the liver the microscopic changes were similar to those occurring in cirrhosis, but in most areas they resembled those seen in congestive liver fibrosis. This case suggested that portal arteriovenous fistula is an important predisposing cause leading to deterioration of liver function.

[Evaluation of the effect of transcatheter arterial embolization as preoperative therapy of hepatocellular carcinoma].

From May 1982 till Nov. 1984, we performed 90 hepatic resection for primary or metastatic liver tumor. Preoperative transcatheter arterial embolization (TAE) was performed on 14 of them. All resected specimens of 14 cases were examined histologically. Effectiveness of TEA was estimated by the ratio of the necrotic area to the whole area of the tumor. Necrosis of the main tumor more than 90% was found in 10 of 14 cases, though necrosis of the daughter nodules was detected in only one of six cases. In hepatectomized patients, two-year-survival of the preoperative TAE cases had not been statistically improved compared with the non-TAE cases.