We applied a platelet activating factor (PAF) antagonist CV-3988 to the acute gastric erosions in rats caused by water immersion and restrained stress to see if PAF is concerned in the acute gastric mucosal lesions (AGML) under stress. CV-3988 inhibited the formation of hemorrhagic gastric erosions by water immersion and restrained stress in a dose-dependent manner. The inhibitory effect of CV-3988 was also elicited in rats given indomethacin under water immersion and restraint stress. CV-3988 20mg/kg inhibited the ethanol-induced gastric erosions of rats. This inhibitory effect was not detereoratd by the depletion of prostaglandins by indomethacin. Our results indicate that PAF is concerned in the formation of AGML under stress.
With the development of genetic engineering, a large amount of recombinant interleukin 2 can be available and adoptive immunotherapy with lymphokine activated killer (LAK) cells can be easily performed. However clinical results with this treatment are not so good in stead of excellent results in vitro. We measured LAK activity in gastrointestinal cancer patients for the basis of this treatment. We could not find any LAK activity correlation between age, types of gastrointestinal cancers, tumor volume and recurrent cases except for high aged recurrent caces which showed lower LAK activity. For the LAK therapy it is one of the problems that over 1×109 cells should be necessary in a short period. In addition contamination must be avoided during the culture. Thus we developed a new high yielding cell culture system. It consists of a culture bag on a rotatar which has two compartments, separated from an outer compartment by a semipermeable membrane. With this system LAK cells can be easily induced without contamination. Furtheremore final cell counts can be 4.4 times more than intial cell counts.
The occupational investigation and the clinical study of patients with pneumatosis cystoides intestinalis (PCI) reported in Japan since 1979 were carried out. Forty three of 65 cases (66.2%) of large intestine type of PCI (pneumatosis cystoides coli, PCC) had had an occupational history of trichloroethylene (TCE) exposure. TCE associated PCC affected predominantly women in the fifth decade, involved mainly left colon, and caused various symptoms, such as abdominal fullness, pain and constipation. These symptoms were found more frequently in TCE associated cases than in cases of other 2 types, secondary and idiopathic PCC. Spontaneous healing and relapse were often recognized in TCE associated PCC, and also in the other 2 types of PCC. Patients with TCE associated PCC had worked during 5.7 years in average in the factories using TCE, where less than 600 employees were working together with them. Ten of 43 patients with TCE associated PCC (23.3%) had been worked in the same 5 factories before the development of the disease.
To investigate the role of reaginic mechanisms in ulcerative colitis, IgE positive and IgG4 positive cells (using immunoperoxidase staining), mast cells (using toluidine blue staining), eosinophils and tissue histamine levels (using fluorometric method with o-phthaldialdehyde) were studied. In active stage, there were significantly more IgE positive and IgG4 positive cells, and eosinophils as well as significantly elevated tissue histamine levels compared with ulcerative colitis in remission and normal controls. In active disease, mast cells were reduced in number suggesting degranulation and releasing histamine into the tissue. Following induction of remission, there was a decrease in the number of IgE positive and IgG4 positive cells, eosinophils and tissue histamine levels and an increase in the number of mast cells. These studies strongly suggested that the reaginic mechanisms were involved in recurrence and aggravation of ulcerative colitis, playing a role as an initiator of inflammation.
To investigate the transport process of secretory immunoglobulin A (sIgA) in active Crohn's disease (CD), colonic specimens were obtained from the site close to the ulcerative or erosive lesions of the patients with CD and were applied to immunohistochemical study for immunoglobulin A (IgA) and secretory component (SC). Light microscopic immunohistochemistry showed that the staining intensity of SC and IgA was increased on the basolateral membrane and at the base of the brush border in the whole mucosal epithelium in CD compared with control subjects. Electron microscopic immunohistochemistry revealed that the transport and secretion of SC and IgA into intestinal lumen were observed in CD as well as control subjects. Furthermore, SC was frequently demonstrated in the interstitium of the lamina propria in CD, possibly released from the basal plasma membrane of the epithelium. The sIgA content of homogenized colonic mucosa was significantly elevated in patients with CD. These results suggested that the transport and secretion of sIgA into intestinal lumen could not be impaired and elevated serum level of sIgA might result from release of excessively produced sIgA in the colonic mucosa in active CD.
The in vitro uptake of bromodeoxyuridine (BrdU) was studied in 17 patients with CAH and 12 patients with L, C, using liver biopsy specimens. The labeling index of BrdU in L, C (2.49±1.90%) was significnatly higher than in CAH (0.93±0.16%). We already reported that the L.I. in portion of H.C.C was highest, and was second highest in noncancerous cirrhotic portion of H.C.C. Considering the above mentioned results, it is likely that L, C is nearer to pre-cancerous state than CAH.
Leukotrienes are chemical mediators which induce allergy and inflammatory reactions. In order to examine the liver metabolism of leukotrienes, we have investigated the existence of leukotrien B4 and C4 (LTB4 and LTC4) in rat liver tissue. Normal rat liver tissue was obtained after laparotomy. LTB4 and LTC4 in bile were assayed by using high performance liquid chromatography and radioimmunoassay. As a result, LTB4 and LTC4 were detected in normal liver tissue in rat. Seven days after intravenous injection of heat-killed Propionibacterium acnes (P. acnes) into normal rat, levels of LTB4 and LTC4 decreased in liver tissue. When both P. acnes and lipopolysaccharide was injected into rat tail vein, LTB4 and LTC4 in rat liver tissue increased. These results suggest that leukotrienes may exist in liver tissue, and may be associated with the induction of liver cell injury.
This study was designed to elucidate the effect of endotoxin (ET) on hemodynamic changes, plasma eicosanoid levels and hepatic adenine nucleotide content in thioacetamide-induced cirrhotic rats (TAA-rats), and to evaluate prophylactic treatment against ET shock. The following results were obtained. 1) In TAA-rats portal blood flow (PBF) and liver blood flow (LBF) decreased significantly in comparison with the control, indicating deterioration of hemodynamics and reduction of blood flow to liver parenchyma. 2) In both groups blood pressure and LBF decreased gradually after ET injection. However, these values in the TAA-rats were lower than those in the control after ET injection; ET injection induced in shock in TAA-rats. 3) Plasma TxB2 levels in both groups were markedly elevated 15 min after ET injection, which was associated with an intermittent reduction in the ratio of 6-keto-PGF1α to TxB2. 4) Hepatic ATP content and energy charge decreased gradually until 120min after ET injection. A significant decrease was observed in the TAA-rats compared to the control. These results suggest that ET exacerbated the impaired liver energy metabolic activity. 5) Pretreatment with 16, 16-dimethyl-PGE2 improved these ET depressed parameters, indicating a prophylactic effect against ET shock and a reduction of mortality.
We investigated the influence of the transcatheter arterial mbolization (TAE) to the cellular and humoral immunity in 19 cases with hepatocellular carcinoma (HCC).1) After TAE, leucocytes, mainly neutrophil increased and lymphocytosis was mild. 2) In HCC, decrease of T4 (helper/inducer T cell) and increase of T8 (suppressor/cytotoxic T cell) were seen and TAE tended to make improve those abnormalities. B1 (B cell) and T11 (Pan T cell) tended to increase after TAE. 3) NK activity and the titer of blastogenesis tended to be reduced after TAE, but those alterations were occurred within the normal range. 4) Immunoglobulin G, A, and M showed high level before TAE and decreased significantly after TAE. 5) Complement (C3c) was low before TAE and the serial changes after TAE was inconsiderable. 6) Immune complex was found to be high before TAE and decreased after TAE significantly. Those changes (1-6) had begun two days after TAE and returned to the pre-treated level one month after TAE.
To evaluate the role of Kupffer cells (KCs) in hepatic fibrosis, KCs isolated from rat liver by a collagenase perfusion method were cultured with and without a stimulant, lipopolysaccharide (LPS). Supernate from the culture of LPS-stimulated KCs contained a fibroblast growth factor (FGF) that caused a significant increase of 3T3 fibroblast DNA synthesis. Furthermore we estimated the possible role of KCs in transient hepatic fibrogenesis induced by acute carbon tetrachloride (CCl4) treated rats stimulated fibroblast proliferation to a greater degree than that of control rats. Enhanced FGF production of KCs occured within 24hr and increased 4-fold at 48hr after injury. These findings suggest that KCs modulate fibroblast growth by releasing a growth factor and play an important role in the progression of hepatic fibrosis.
Biopsied liver specimens from 69 patients carrying HBsAg positive chronic hepatitis in whom 27 patients were treated with interferon because of HBeAg positive were used to investigate prognostic significance of HBcAg in chronic hepatitis. There was a quantitative correlation between HBcAg in liver tissue and serum HBV markers (HBeAg, DNA polymerase, HBV-DNA). In addition, expression of HBcAg in the liver was characteristic in accordance with histological activity of chronic hepatitis. That is, cytoplasmic localization of HBcAg was observed predominantly in the livers from chronic active hepatitis. Effect of interferon was seen in the patients whose livers showed lesser quantitative but cytoplasmic distribution of HBcAg. Therefore, it is very helpful for assuming the prognosis of chronic hepatitis and indication of interferon therapy to study the types of HBcAg distribution in the livers.
We studied the changes of serum PH in hepatocellular carcinoma (HCC) and gastrointestinal cancer in terms of liver metastasis and also examined a possible mechanism of serum PH elevation from stand point of immunohistochemical localization of tissue PH. 1) The activity of serum PH was elevated according to the growth of HCC, and decreased with effective anticancer therapy. 2) Serum PH remained within normal range in almost all cases of various gastrointestinal cancer without liver metastasis, however the level increased remarkably with liver metastasis. 3) Immunohistochemical PH was stained at the marginal HCC cells, and hepatocytes bordering both HCC and metastatic liver cancer. And primary site in various gastrointestinal cancer was stained negative. The elevation of serum prolyl hydroxylase may be derived from those immunohistochemically stained cells with PH.
Serum CA125 levels in 33 patients with hepatocellular carcinoma were studied and compared with serum levels of AFP, CA19-9 and CEA. Mean (±SEM) serum CA125 levels and positivity in patients with hepatocellular carcinoma were 406±97U/ml and 75%, and significantly higher than those in liver cirrhosis. Elevation of CA125 was found in patients with ascites, and/or with advanced disease stage. The sensitivity, specificity and accuracy of CA125 in hepatocellular carcinoma were not different from those of AFP, but significantly higher than those of CA19-9 and CEA. However, in the patients with hepatocellular carcinoma without ascites, those of CA125 were significantly lower than those of AFP. We conclude that serum CA125 is useful in diagnosing the stage of hepatocellular carcinoma.
Primary culture or mucosal epithelial cells of rabbit gallbladder was attempted in collagen gel matrix and their morphological features were examined for up to six weeks. Singly isolated epithelial cell and epithelial cell clots began to proliferate and various sized cysts were formed within them. These cysts were lined by cuboidal-columnar epithelia and mucus was secreted into the cystic cavities. Mucus was also demonstrated within cytoplasmic vacuoles. Transmission electron microscopy showed good preservation of the original gallbladder epithelia; cultured epithelia showed well formed microvilli facing cyst, desmosomes, tight junction and interdigitation. In their cytoplasm there were small-sized mitochondria, numerous intermediate filaments, well developed Golgi's apparatus and mucus droplets. Cytokeratin was demonstrated by immunofluorescence method. This culture method seems valuable for the in vitro study of gallbladder epithelia.