Shift of hepatitis E virus RNA from hepatocytes to biliary epithelial cells during acute infection of rhesus monkey.

Hepatitis E virus (HEV) has been considered to be the major cause of enterically transmitted non-A, non-B hepatitis in developing countries. However, little is known about viral replication and localization in the liver. The aim of this study was to examine the distribution of HEV-infected cells in experimentally infected animals. Seven captured wild rhesus monkeys were inoculated intravenously with faecal extract derived from a Myanmar strain of HEV. Animals were killed at different time-points of clinical illness: during early infection, during prehepatitis with viral-like particles in bile, during acute hepatitis and during convalescence. Intrahepatic localization of HEV was analysed using non-isotopic thymine dimer in situ hybridization (NITDISH). Both plus and minus strands of HEV RNA were found in hepatocytes during the early infection period. Staining in the submembranous cytoplasmic region of hepatocytes was observed. In the prehepatitis period, both plus and minus strand HEV RNAs appeared in the canalicular side of isolated bile epithelial cells. Subsequently, HEV RNA became universally distributed in the cytoplasm of medium-size bile epithelial cells. After recovery, HEV RNA disappeared.

Chronic Japanese schistosomiasis and hepatocellular carcinoma: ten years of follow-up in Yamanashi Prefecture, Japan.

In a preliminary study carried out in the study area we found that 19.1% (173/907) of patients with chronic liver disease and 51% (35/68) of hepatocellular carcinoma cases were infected with Japanese schistosomiasis. Analysis of data from 571 autopsies revealed a similarly high incidence of schistosomiasis among cases of hepatoma and other liver diseases. A prospective case-control study conducted over 10 years showed that hepatoma developed in 5.4% (26/484) of chronic schistosomiasis cases and in 7.5% (23/307) of patients with chronic liver disease (hepatitis, cirrhosis, etc). The difference was not statistically significant (P = 0.228). A high incidence of hepatitis C virus (HCV) antibody (HCVAb) was found in the schistosomiasis group (36.5%; 95% CI = 44.9-28.1%) and in the chronic liver disease group (56.0%), 39% of whom had chronic hepatitis (P = 0.028). Various factors that might have contributed to the development of hepatoma and schistosomiasis were investigated, but no evidence of a significant correlation between schistosomiasis and hepatoma was found. The high incidence of HCVAb was considered to have been responsible for the development of hepatocellular carcinoma in chronic schistosomiasis patients. The role of HBV infection in the development of hepatoma in schistosomiasis patients was not confirmed after an assay for HCVAb was included in the study.

Genotyping of the CYP1A1 and GSTM1 genes in esophageal carcinoma patients with special reference to smoking.

BACKGROUND: Aromatic hydrocarbons, including benzol[a]pyrene, in tobacco smoke first require metabolic activation by phase I enzymes, cytochrome P450s (CYP450s), and then are subjected to detoxification by phase II enzymes, the glutathione-S-transferases. A high risk lung carcinoma group has been reported to have specific polymorphisms of the cytochrome P450 (CYP1A1) gene and the glutathione-S-transferase (GSTM1) gene. In this study, the authors investigated whether such genotypes were also risk factors for esophageal carcinoma. METHODS: Subjects were comprised of 89 esophageal carcinoma patients and 137 noncancer controls. Forty-nine of the patients and 60 of the control subjects were smokers. Genotypic studies of both CYP1A1 and GSTM1 were performed in the cancer tissues of all 89 patients. Genotypes of peripheral blood leukocytes taken from the control subjects were also determined. Genotypes of the CYP1A1 and GSTM1 genes were determined by the polymerase chain reaction. RESULTS: Patients who were heavy smokers with the genotypes Val/Val (V/V) for CYPIAI and the combined genotype of V/V for CYP1A1 and GSTM1- were a statistically high risk group compared with control subjects (P < 0.01, chi-square = 10.6 vs. P < 0.01, chi-square = 11.0). The association of V/V for CYPIAI with a smoking index > or = 600 in esophageal carcinoma patients was estimated at 6.63 (95% confidence interval [CI], 1.86-23.7). The association of combined genotypes of V/V of CYP1A1 and GSTM1 with a smoking index > or = 600 in esophageal carcinoma patients was estimated at 12.7 (95% CI, 1.97-81.8) CONCLUSIONS: Specific genotypes of the CYP1A1 and GSTM1- genes are related to the incidence of esophageal carcinoma, especially in heavy smokers.

Immunohistochemical studies of vascular volume and proliferative activity in squamous cell carcinoma of the esophagus.

For immunohistochemical investigation and clarification of the relationship between the vascular volume in esophageal carcinoma and the proliferative activity of its tumor cells, we examined surgical specimens of squamous cell carcinoma (SCC) of the esophagus from 15 patients. The vascular volume was evaluated by immunostaining for platelet-endothelial cell adhesion molecule-1 with monoclonal antibody JC70, and the proliferative activity of the carcinoma cells was evaluated by immunostaining for proliferating cell nuclear antigen (PCNA) with antibody 19A2. The ratio of the vascular area to the tumor area and the labeling index (LI) for PCNA in the carcinoma cells was then calculated. The antibody JC70 was useful for immunohistochemically detecting blood microvessels in esophageal carcinoma. The vascular volume, expressed as the ratio mentioned above, was higher in well- and moderately differentiated SCCs than in poorly differentiated SCC (P < 0.01), and the PCNA LI did not depend on the degree of differentiation. However, there was a significantly inverse relationship between the ratio of the vascular area to that of carcinoma and the PCNA LI of the carcinoma cells (P < 0.01). These findings show that angiogenesis is greater in esophageal carcinomas with little proliferative activity.

Expression of cytochrome P450s and glutathione S-transferases in human esophagus with squamous-cell carcinomas.

In order to clarify the expression of cytochrome P450 and glutathione S-transferase in human esophagus, 41 samples of human esophagus with squamous-cell carcinoma were investigated by immunoblot analysis and enzyme assays. Cytochrome P450 1A2/1 was clearly expressed in microsomes, and the amount in samples with tumorous tissue was significantly greater than that in samples without tumourous tissues or in liver; cytochrome P450 2B6 and 3A4/3 were expressed polymorphically. Aryl hydrocarbon hydroxylase activity was detected in microsomes and was greater in samples from smokers than non-smokers. Patients who both smoked and drank alcohol, however, had activity similar to that of patients without these habits. Glutathione S-transferase M1 and A1/2 protein existed polymorphically in cytosol, and glutathione S-transferase P1-1 was detected in all samples. The frequency of expression of the glutathione S-transferase A1/2 protein was greater in patients with M1 protein than in those without; no difference in the expression was seen for glutathione S-transferase P1-1. Neither smoking nor drinking influenced the expression or activity of glutathione S-transferase. Our data support the idea that some carcinogens can be directly activated or inactivated in human esophageal epithelium.

Clinicopathologic characteristics and postoperative outcome in Japanese and Chinese patients with thoracic esophageal cancer.

We evaluated the clinicopathologic findings and surgical results of 140 patients with thoracic esophageal cancer treated at Shinshu University, Japan (Shinshu group), and compared them with those from 1164 patients treated at Hebei Medical College, China (Hebei group) to determine if the two groups showed any differences. The Shinshu group had significantly higher incidences of elderly patients (>70 years of age), male patients, and tumors located at the lower esophagus (p < 0.01). In the Hebei group, although the depth of tumor invasion was more advanced, the incidence of nodal metastasis was significantly lower (p < 0.01). Operative death and postoperative complications were more frequent in the Shinshu group. Comparison of the postoperative survival curves revealed significantly longer survival of patients with pT2 or pT3 tumor in the Hebei group (p < 0.01), but there were no significant differences between the two groups when the lesions were classified by pTNM stage. This study demonstrated several differences between the patients in the two areas in regard to the clinicopathologic characteristics of thoracic esophageal cancer. The most important characteristic of the esophageal cancer in the Hebei group appears to be the low incidence of nodal metastasis.

Assessment of proliferative activity of glandular cells in hyperfunctioning parathyroid gland using flow cytometric and immunohistochemical methods.

The proliferative activity of epithelial cells in hyperfunctioning parathyroid glands was estimated by flow cytometric and immunohistochemical procedures. A total of 30 parathyroid glands, 29 hyperfunctioning glands, and 1 normal gland were studied. The pathology of the 29 glands was determined to be hyperplasia in 19 and adenoma in 10. The S-phase cell population was expressed in terms of the S-phase fraction (%SPF) calculated from the histogram by DNA flow cytometry and in terms of the BrdU immunostaining labeling index (BrdU LI). Cells in all stages of the cell cycle were studied by Ki-67 immunostaining and expressed in terms of the labeling index (Ki-67 LI). Both BrdU LI and Ki-67 LI values were low, ranging from 0% to 0.58% and from 0.21% to 2.62%, respectively. The BrdU LIs were lower than the Ki-67 LIs, ranging from one-sixth to one-twelfth of the values depending on the disease. There were significant correlations between the two indices (p < 0.001). The %SPF determined by flow cytometry was consistently higher than both the BrdU LI and the Ki-67 LI. This discrepancy cannot be explained precisely, and further improvements are required for the flow cytometric analysis of %SPF. The cell cycle study by BrdU and Ki-67 immunohistochemistry suggested that the glandular cells of the hyperfunctioning parathyroid were characterized by low proliferative activity. No evidence of rapid cell turnover rate assumed from the flow cytometric study could be observed in the hyperfunctioning parathyroid gland.

[Immunohistochemical expression of sialyl-Lewis antigens in lung cancer].

We investigated the immunohistochemical expression of three carbohydrate antigens, sialyl-Lewisa, sialyl-Lewisx and sialyl-Lewisx-i, in human lung cancer tissues using monoclonal antibodies, 2D3, SNH3 and FH6, respectively, and compared the expression rate of these three antigens with clinical and pathologic findings. The expression rate of all the three antigens in adenocarcinoma was higher than that in squamous cell carcinoma and that of sialyl-Lewisx was highest in adenocarcinoma. Sialyl-Lewisx antigen was expressed in all cases of positive nodal metastasis or postoperative distant metastasis in adenocarcinoma. In squamous cell carcinoma, however, there was no relationship between the expression rate of sialyl-Lewisx antigen and nodal or distant metastasis. These results suggest that expression of sialyl-Lewisx antigen in adenocarcinoma of the lung may be correlated with nodal or distant metastasis.

Management of tracheal wall resection for thyroid carcinoma by tracheocutaneous fenestration and delayed closure using auricular cartilage.

BACKGROUND: In the case of advanced thyroid carcinoma invading the trachea, surgical procedures for tracheal resection are selected in relation to the degree of invasion. We created a tracheocutaneous fenestration that was later closed with a free auricular cartilage autograft. We present a simple and effective method for permanent closure of tracheostoma. METHODS: The surgical procedure for delayed closure consists of three steps: (1) preparing the hinge flap with additional undermining of the subcutaneous layer and the strap muscles; (2) removing a portion of the conchal cartilage from the auricle, which is easily performed with a semicircular skin incision along the anthelix of the ear; and (3) placing the conchal cartilage with its convex surface upward just over the sutured hinge flap layer. RESULTS: Under local anesthesia, we successfully employed this reconstructive procedure for 5 patients. Unfortunately, however, in one patient, we had to reopen the closed tracheocutaneous fenestration on the second postoperative day because of an acute problem that was independent of the closure surgery. Four patients remain in satisfactory condition for more than 2 years since the closure of tracheostoma. CONCLUSIONS: The auricular cartilage free graft is a relatively simple procedure which supplies tracheal wall skeletal support for the permanent closure of the tracheocutaneous fenestration.

Management of chylothorax after pulmonary resection.

BACKGROUND: Conservative management with intrapleural drainage and total parenteral nutrition (TPN) has been the first choice of treatment for postoperative chylothorax. With this approach, however, it usually takes several weeks for the chylothorax to resolve and it is sometimes unsuccessful. In this study, we reviewed seven patients who had chylothorax develop after pulmonary resection for primary carcinoma of the lung. STUDY DESIGN: The patients were treated according to a "one-week trial" that consisted of one week of observation with intrapleural drainage and maximum parenteral nutritional support followed by operative intervention if the effect of the conservative therapy was not adequate. When the chylous leak was decreased to less than 100 mL/day or less than 15 percent of the maximum daily drainage volume after the "one-week trial," the conservative management was continued for two more weeks. After observation for three weeks, oral intake was begun and a final evaluation of the treatment was made. RESULTS: One patient did not consent to the "one-week trial" and underwent operative treatment on the third postoperative day. Two patients had chylous leaks less than 100 mL/day or less than 15 percent of the maximum daily chylous leak after one week observation. Conservative management with TPN was continued in these patients for two more weeks and operation was performed in one on the 20th day and in the other on the 22nd postoperative day. The remaining four patients underwent operative treatment on the seventh or eighth postoperative day. All of the operations for chylothorax were successful, and chest tubes were removed promptly. These results show that operative management of chylothorax was reliable and safe. The "one-week trial," however, offered few advantages in determining the therapeutic strategy for postoperative chylothorax.

Histopathological features of atrophic thyroiditis with blocking type-TSH binding inhibitor immunoglobulins.

To investigate the histopathological features of atrophic thyroiditis (AT) with blocking type-TSH binding inhibitor immunoglobulins (TBII), the present morphological observations were carried out employing additional immunohistochemical procedures. Moreover, these were compared with examples of goitrous Hashimoto's thyroiditis showing negative TBII (HT). There exist apparent differences between AT and HT. In particular, significant follicular atrophy with epithelial flattening including decreased positive staining of the follicular epithelial cells for thyroglobulin in AT was characteristically observed. These results suggest that the mechanism for the development of hypothyroidism in AT with blocking type-TBII might be due to suppression of thyroid cell function through the inhibition of endogenous TSH stimulation by the blocking antibody with subsequent epithelial degenerative destruction.

Thyroid abnormalities among children in the contaminated area related to the Chernobyl accident.

Gradual increases of various radiation-related diseases including thyroid cancer following the Chernobyl accident have been recently reported. We have carried out systematic thyroid surveys in school children aged 10-15 years in the highly contaminated area and compared the results with a similar survey in a nonaffected neighboring area as a control. Systematic examinations of the thyroid gland were performed in 888 schoolchildren (408 males and 480 females) in the districts of Chechelsk city with 5 to over 40 Ci/km2 of radiation level for 137Cs. In the control area (Bobruisk city), 521 children with the same age range (229 males and 292 females) were examined. Thyroid surveys were carried out as follows: palpation, ultrasonography, blood examinations of thyroid function, and measurements of daily urinary excretion of iodine. Certain thyroid abnormalities were observed in the high radioactive fallout area more frequently than in the control region. In particular, the prevalence of multiple micronodular lesions in diffuse goiter in the contaminated area was significantly higher than in the control area. However, endemic goiter due to iodine deficiency was present in both districts. The environment factor of iodine deficiency might have resulted in the early occurrence of ultrasonographic thyroid abnormalities attributed to radioactive contamination.

Evaluation of imaging examination for hepatic invasion of carcinoma of the gallbladder and postoperative patient outcome.

BACKGROUND: Hepatic invasion is an important factor in the selection of an operative procedure for patients with carcinoma of the gallbladder. Imaging modalities to detect the hepatic invasion were evaluated for accuracy, and postoperative patient outcome was analyzed. STUDY DESIGN: Thirty-six patients who were treated for carcinoma of the gallbladder at our institution between 1984 and 1993 were studied. The imaging results of ultrasonography (US), computed tomography, and angiography were compared with histologic findings regarding intrahepatic invasion of the carcinoma. RESULTS: Histologic examination of the operative specimens removed from 21 patients revealed that the detection of hepatic invasion with preoperative US was superior to that with the other two modalities. The preoperative US showed a diagnostic value that was almost the same as that of the intraoperative US. The postoperative patient outcome was not satisfactory, although extended radical operation was performed using image guidance in the 21 patients. The patients died as a result of nodal recurrence rather than hepatic recurrence. CONCLUSIONS: Preoperative US should be performed initially for detecting hepatic invasion from carcinoma of the gallbladder and can be substituted for intraoperative US. Considering the operative treatment for patients with carcinoma of the gallbladder, lymph node dissection is as important as the extent of hepatectomy.

The safe and complete removal of a carotid body tumor with elements suggestive of a malignant potential by employing an intraluminal shunt: report of a case.

The carotid body tumor is a relatively rare neoplasm arising from the intercarotid paraganglions. We report herein the case of a patient with a Shamblin group 2 type carotid body tumor for which safe and successful removal was facilitated by the insertion of an intraluminal shunt. The patient was a 54-year-old man who presented with a spherical elastic mass, 4.8 x 3.7 cm in size, on the right anterior aspect of his neck, which was diagnosed as a carotid body tumor by diagnostic imaging. Because the tumor was suspected to be malignant based on the operative findings, which included invasive adhesion to the carotid artery, an intraluminal shunt was inserted, allowing for safe and complete removal. Histologically, this tumor showed malignant potential with sporadic mitoses and incomplete capsular invasion. Thus, it is recommended that an intraluminal shunt be employed for the removal of a carotid body tumor when it is found to be tightly adherent to the carotid artery during the operation.

The influence of excess body weight on the surgical treatment of patients with gastric cancer.

Sixty-two overweight gastric cancer patients were compared with 201 normal-weight patients to clarify the influences of excessive weight on the surgical treatment of gastric cancer. The frequencies of hypertension and diabetes mellitus were significantly higher in the overweight group (P < 0.01), but no pathologic differences in the resected tumor were found between the two groups. The operative times were longer (P < 0.01) and the number of lymph nodes extirpated and examined was smaller (P < 0.01) in the overweight group. The incidence of postoperative complications was not higher in the overweight group. The postoperative survival rate of patients with nodal metastasis was statistically lower in the overweight group (P < 0.05). Regarding the causes of death in patients with nodal metastasis, 61.1% of overweight patients and 43.8% of normal-weight patients died of recurrence of gastric cancer. In conclusion, surgical treatment of overweight patients with gastric cancer was found to be technically more difficult and the prognosis of such patients with nodal metastasis may thus be worse than that of their normal-weight counterparts.

Preoperative intraperitoneal chemotherapy for gastric cancer, with special reference to delayed peritoneal complications.

Preoperative intraperitoneal (IP) chemotherapy was performed in 23 patients with gastric malignancies to inhibit peritoneal recurrence. Cis-diamminedichloroplatinum (CDDP) and mitomycin C (MMC) were administered intraperitoneally 3 days prior to surgery, at which time a very viscid peritoneum and mucinous intraperitoneal fluid were found in 100% and 83% of the patients, respectively. Inflammatory changes were microscopically observed in the subserosal layer of the resected stomachs and in the intraperitoneal fluid, but degenerative changes characteristic of cancer cells could not be seen. The 3-year survival rate of the stage III patients was 55.6%, and peritoneal recurrence was found in three of six patients with recurrence. Extensive adhesions were found in eight patients (34.8%) as a delayed peritoneal complication, and chronic bowel obstruction resulting from the adhesion developed in five patients (21.7%). Thus, we conclude that the administration of this IP chemotherapy demonstrated no definite antitumor effects or survival benefits, but was frequently associated with delayed peritoneal complications.

Lower esophageal sphincter pressure after subtotal gastrectomy and postoperative reflux esophagitis.

Although reflux esophagitis after subtotal gastrectomy has been noticed by surgeons, the mechanism of its development is obscure. This study was undertaken with the aim of clarifying the role of the lower esophageal sphincter in the development of this abnormality. Manometric studies were carried out on 42 patients with gastric cancer, and on 19 with cholelithiasis. The lower esophageal sphincter pressure was measured using a catheter tip pressure transducer and a rapid pull-through technique, and the results presented as the mean of three measurements. The technical error of this experiment was estimated to be within 2 mmHg by a study of the cholecystectomy patients. After gastrectomy, the lower esophageal pressure decreased in 17 patients, increased in 4 and remained unchanged in 21. A more pronounced decrease in the lower esophageal sphincter pressure was found after Billroth II. Clinical evaluation of the 42 patients revealed symptoms of postgastrectomy regurgitation in 10. Preoperatively, these 10 had lower values of the lower esophageal sphincter pressure followed by a more marked postoperative decrease, as compared with the patients with no regurgitation symptoms. Oral administration of a test meal revealed regurgitation after subtotal gastrectomy. This study suggests that a low value of the pre-operative lower esophageal sphincter pressure, a marked decrease in the pressure after gastrectomy, and Billroth II anastomosis, may be factors that predispose to regurgitation.

Protective effect of the thromboxane A2 receptor antagonist ONO 3708 on ischemia-reperfusion injury in the dog liver.

BACKGROUND: Prostaglandins are widely known to have cytoprotective effects in a variety of conditions. Thromboxane A2 has the opposite effect of prostaglandins. In this study the effects of the thromboxane A2 receptor antagonist ONO 3708 on ischemia and subsequent reperfusion in the dog liver was evaluated. METHODS: Mongrel dogs weighing from 10 to 15 kg were divided into three groups: a control group, a group with induced liver ischemia and subsequent reperfusion, and a group that received ONO 3708 and then underwent induced liver ischemia and subsequent reperfusion. Liver ischemia was induced by the Pringle procedure for 60 minutes. The concentrations of total free amino acids, aromatic amino acids, and branched-chain amino acids in blood taken from the portal and hepatic veins were examined before and after the Pringle procedure in the latter two groups and at the corresponding points in the control group. RESULTS: Disturbances in amino acid metabolism in the liver occurred 5 minutes after the declamping in the ischemic group, and prostaglandin I2 and thromboxane A2 levels and lipid peroxide production, were increased. In contrast, hepatic amino acid metabolism was unchanged, and prostaglandin I2 and thromboxane A2, and lipid peroxide production, were normalized in the group that was treated with ONO 3708. CONCLUSIONS: ONO 3708 appears to protects hepatic tissue from ischemia-reperfusion injury through free-radical scavenging, by increasing prostaglandin I2 levels, and by decreasing thromboxane A2 production.

[Combination neoadjuvant therapy with continuous intra-arterial CDDP infusion and radiotherapy for locally advanced breast cancer].

Neoadjuvant therapy combined with intra-arterial continuous infusion of CDDP and external radiation was evaluated in a patient with locally advanced breast cancer. Although only MR was observed on the primary tumor, PR was found in several metastatic lymph nodes and daughter nodules. All tumor markers elevated before treatment decreased to normal ranges only by this therapy, and histological damage was also recognized remarkably with a high concentration of free Pt (8.89 micrograms/g) in cancer tissues. Since no severe side effects appeared during the present therapy, extended radical mastectomy with wide skin resection could be performed following additional intra-arterial chemotherapy using 5-FU, epi-ADM and MMC. From this experience, it was suggested that CDDP was an useful drug in the chemotherapy for breast cancer.