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Bile duct injury can occur more frequently during laparoscopic cholecystectomy than in open cholecystectomy. Three cases of common bile or hepatic duct injuries occurred in a series of eighty laparoscopic cholecystectomies; In case 1, the common bile duct was misidentified as the cystic duct. In case 2, bile peritonitis occurred on the fourth postoperative day caused by necrosis of the common hepatic duct involving the cautery surrounding it. In case 3, a bile leak occurred due to an incision at the confluence of the cystic and common duct. Dissection of the cystic duct at the infundibulum of the gallbladder, blunt dissection of the Calot's triangle, the handling of clips with special attention for safety were thought to be necessary in order to lower the risk of bile duct injury. Preoperative endoscopic retrograde cholangiography (ERCP) is recommended to avoid bile duct injury.

Reduced indocyanine green (ICG) uptake is one of the functional changes of human hepatocellular carcinoma (HCC). To clarify the mechanisms of loss of ICG uptake, and determine which subunit of glutathione-S-transferase (GST), alpha or pi, plays a role in ICG transport in hepatocytes, an experimental HCC model was developed that used nodules induced by 2-acetylamino-fluorene (2-AAF) administration. Many of the ICG stained nodules, which consisted of benign and borderline lesions, were GST-alpha positive. However, the percentage of GST-alpha positive cells tended to decrease according to the disappearance of ICG staining in the process of hepatocarcinogenesis. HCCs unstained by ICG were also GST-alpha negative. GST-pi, not detected in normal rat hepatocytes, appeared in an earlier stage of hepatocarcinogenesis before the disappearance of GST-alpha, and was not observed in HCCs. No significant relationship between ICG staining and GST-pi was recognized. These results suggest that GST-alpha synthesis is disturbed in the process of hepatocarcinogenesis and results in loss of ICG uptake in HCCs, and also indicate that GST-pi may be useful for early diagnosis of preneoplastic hepatocytes showing no roles in ICG transport.

The negative strand RNA of hepatitis C virus, supposed to be a replicative intermediate of the virus appears to indicate viral replication. In this study, we detected the negative strand RNA by using reverse transcription-polymerase chain reaction with RNase A digestion to degrade the remaining positive strand genomic sequence of the virus after complementary DNA (cDNA) synthesis. In vitro transcribed positive-stranded mutant RNA was not detected by this method. Sample sera and liver tissues of 16 patients with chronic hepatitis C virus infection (liver fibrosis, 1; chronic hepatitis, 13; liver cirrhosis, 2) were analysed for negative strand RNA of hepatitis C virus. The negative strand RNA sequence was detected in 15 (93%) of 16 liver tissues and in 11 (78%) of 14 sera. The study demonstrated that negative strand RNA of hepatitis C virus in serum and liver tissue could be specifically detected.

Three cases of successful pregnancies in renal transplant recipients who had undergone transplantation in the Okayama University Medical School Hospital are reported. Two of the women had received an organ from a living relative and one woman received a cadaveric organ graft. These patients, aged 28-37 at the time of the delivery, had received their transplants 2-5 years prior to their conception. The periods of gestation ranged between 35 and 40 weeks. The weight of the babies at birth ranged from 2,380g to 2,500g and the apgar score at 1 min was 8 or 9. None of the infants showed any congenital abnormalities. Lower-segment cesarean section was performed in all of three cases. Serum creatinine levels, an indicator of renal graft function, did not deteriorate during the pregnancy or after delivery. Although further work is needed to solve problems regarding pregnancy in renal transplant recipients, these results encouraged us to meet their hope for a baby.

In order to clarify the mechanism of retinal tissue damage in diabetes mellitus, the effects of the inhibition of aldose reductase on the pathologic changes in the retina of streptozotocin-induced diabetic (STZ-diabetic) rats were examined histologically and histochemically. The STZ-diabetic animals were maintained with and without peroral administration of an aldose reductase inhibitor, SNK-860, and their retinas were examined microscopically after 12 months. Several abnormal changes observed; folding and edema in the retina, loss of pericytes in the retinal capillary walls, and thickening of basement membranes in the retinal capillaries, were significantly inhibited by SNK-860. Some of these changes were similar to those that had been previously noted in diabetic and galactosemic rats. These data suggest that the enhanced polyol metabolism may be involved in the diabetic changes of the retina.

We evaluated the viability of the cadaver lung and the effect of lung inflation with 100% oxygen using a canine allotransplantation model. Donor animals were killed by potassium chloride (KCl) injection and were kept at room temperature until lung extraction. The animals were divided into the following 3 groups: group 1 (n = 6) in which the donor lungs were retrieved 2h after sacrifice, group 2 (n = 6) in which the donor lungs were retrieved 3h after sacrifice, and group 3 (n = 6) in which the donor lungs were retrieved 3h after sacrifice as in group 2 except that they were kept inflated for 3h with 100% oxygen using a double lumen endotracheal tube. Heparin was not given and lungs were not flushed with preservation solution. After left lung transplantation, the transplanted lung function including gas exchange and pulmonary hemodynamics was assessed for 6h by ligating the right pulmonary artery of the recipient animals. All 6 animals in groups 1 and 3 survived for 6 h with excellent lung function. Only 2 of 6 animals in group 2 survived for 6h with poor lung function. These results led us to conclude the following: a) the cadaver lung kept at room temperature for 2h might be available for lung transplantation, and b) when the cadaver lung is inflated with 100% oxygen, the length of safe ischemic time could be prolonged up to 3h.

The effects of long-term glucocorticoid therapy on chemical mediator and cellular reaction in the airways were examined in 69 patients with bronchial asthma. The histamine release induced by Ca ionophore A23187 from cells in the bronchoalveolar lavage (BAL) fluid of atopic asthmatics was significantly lower in the subgroup with steroid-dependent intractable asthma (SDIA) than in non-SDIA patients (p < 0.05). In contrast, histamine release in nonatopic SDIA patients did not differ from nonatopic non-SDIA patients. The release of leukotriene C4 (LTC4) was significantly lower in atopic patients with SDIA (p < 0.02). However, there was no significant difference in LTC4 release between nonatopic patients with SDIA and without SDIA. The proportion of BAL lymphocytes was significantly lower in atopic patients with SDIA than in those without it (p < 0.05), although there was no significant difference between the nonatopic patients with and without SDIA. These results show that glucocorticoids affect humoral and cellular events in the airways of atopic asthmatics more than in those of nonatopic asthmatics.

Epiphyseal growth cartilage of the femoral head obtained from Wistar rats was investigated after fixation by a rapid-freezing and freeze-substitution. Liquid helium was used in order to achieve a fast cooling rate without ice-crystal damage during the rapid freezing. Use of the rapid-freezing and freeze-substitution procedure provided better ultrastructural preservation of the chondrocyte than conventional chemical fixation methods. This procedure allowed a more reliable approach to electron probe analysis. X-ray microanalysis of the specimens confirmed that calcium is not detected in the initial matrix vesicles as a result of the freezing process. The results suggest that calcium release from precipitates occurs in the free state without any detectable formation of hydroxyapatite at the initial stage of calcification and that calcium is not tightly bound to the matrix vesicles.

A total of 124 lesions from 1 to 6cm in diameter, including 31 cavernous hemangiomas, 32 metastases and 61 hepatocellular carcinomas (HCC) were analyzed to study the usefulness of magnetic resonance imaging (MRI) at 0.5 Tesla to differentiate focal hepatic lesions on the basis of qualitative criteria. Each focal hepatic lesion was assessed for shape, internal architecture and signal intensity relative to normal liver parenchyma. While all cavernous hemangiomas and metastases except one lesion could be detected, detection rate of HCC was significantly inferior to that of the other two diseases. A tumor capsule and a hyperintense focus on T1-weighted images were demonstrated in only HCC lesions in strong contrast with the other two diseases; however, metastases with slow-growing characteristics or subacute hematoma may appear as similar images. Cavernous hemangiomas appeared markedly hyperintense on T2-weighted images in 23 of 31 lesions, but one metastasis and one HCC had similar images. A multivariate analysis of several MRI resulted in the following mean discriminant scores: cavernous hemangioma, -1.2652; metastasis, 0.1830; and HCC, 0.7138. It appeared to be possible to differentiate the three diseases with 84.4 percent accuracy.

We report second malignant neoplasms which developed between 7 and 19 years after treatment in 3 pediatric patients with osteosarcoma. Two patients had been treated with only surgery, and another patient had been treated with a combination of surgery with chemotherapy and radiation therapy for primary lesions. Pediatric patients with osteosarcoma, in particular, require careful long-term follow-up to monitor not only metastases but also development of second malignant neoplasms.

The aim of this study is to obtain data for improving a training program for patients with diabetes mellitus. One hundred eighty-seven patients with non-insulin dependent diabetes mellitus were tested with 20 questions about their knowledge for self-management of diabetes mellitus. Then to draw out factors in their personal backgrounds relating to their correct answers, multiple regression analyses were conducted. As a result, four factors showed significant differences in the following order: Educational careers > ages > duration of disease > socioeconomic strata. The results of the present study have shown for the first time, that these four factors closely concern patients to acquire the necessary knowledge for their self-management of the disease. In addition, this study has raised some fundamental problems regarding the training program for patients: how education should be given to patients.

To confirm the formation of Charcot-Leyden crystals (CLC) in basophils, we observed basophils in sputum and peripheral blood. Sealed slide and suspension culture methods were used to observe the process of CLC formation in peripheral blood basophils and eosinophils under electron microscopy. CLC formation was observed in basophils and eosinophils, and was found to be augmented by sealed slide method. A temperature of 4 degrees C was better than 37 degrees C for promoting the formation of crystals. There was no correlation between the degranulation of these cells and the formation of CLC after stimulation with anti-IgE or anti-IgG antibodies. CLC were initially detected in the cytoplasmic granules of basophils where they continued to enlarge. No CLC were identified in mast cells under any conditions studied. These findings confirm that CLC in sputum are not exclusive to eosinophils and that CLC appear to be present in basophil-rich sites under the cell damage.

A newly introduced compound, EPC-K1, represents a phosphate diester linkage of vitamin E and vitamin C. The effect of EPC-K1 on the reperfusion injury was evaluated in a heterotopic cardiac transplantation model using syngenic combination rats. Prior to the warm ischemia, 12mg EPC-K1/kg was administered intravenously to donor rats. After 15 min of warm ischemic time, hearts were harvested and perfused with 4 degrees C saline. After completion of the transplantation, recipient rats were also treated with intravenous 12 mg EPC-K1/kg, before reperfusion. Saline was used instead of EPC-K1 for both donors and recipients in the control group. On the 7th post-transplantation day, graft survival was 7 out of 8 in EPC-K1 group, versus 1 out of 9 in the control group (p < 0.001). Thiobarbituric acid-reactive substance levels in the recipient serum, three hours after reperfusion, were significantly limited, in the group in which EPC-K1 was administered only to donors. But it was not possible to clarify whether the effect of EPC-K1 is primarily at the donor or recipient levels at this time. These results indicate that EPC-K1 may reduce reperfusion injury after cardiac transplantation. This beneficial effect may be mediated by the hydroxyl radical scavenging properties of EPC-K1.

A sustained release system for interleukin-2 (IL-2), and IL-2 mini-pellet (IL-2 mp), was developed by fusing IL-2 into a needle shaped collagen. Serum concentration of IL-2 after a single subcutaneous injection of the IL-2 mp into C57BL/6 mice remained elevated longer than after an injection of aqueous IL-2. IL-2 in the serum became undetectable by 6h after a subcutaneous injection of 1 x 10(6) unit of IL-2 in phosphate-buffered saline (PBS). In contrast, after a single subcutaneous injection of IL-2 mp containing the same amount of IL-2, the concentration of IL-2 increased to its maximum at 6h after injection, then began to decrease gradually. IL-2 was detected even on the third day after a single subcutaneous injection of one IL-2 mp. Augmentation of NK activity and generation of IL-2 activated killer cells were observed in the spleen from day 1--day 3 after a single subcutaneous injection of IL-2 mp into C57BL/6 mice. This activation was not observed following a single subcutaneous injection of the same amount of IL-2 in PBS. Adoptive immunotherapy by a single subcutaneous injection of IL-2 mp followed by intravenous injections of in vitro cultured IL-2 activated killer cells showed better results in decreasing the number of metastases of Lewis lung carcinoma in C57BL/6 mice than immunotherapy using IL-2 solution.(ABSTRACT TRUNCATED AT 250 WORDS)

A 56 year-old rectal cancer patient who developed a chronic rectoabdominocutaneous fistula postoperatively was treated with fibrin clot, and the fistula healed completely. Occlusion of chronic postoperative fistulas with fibrin clot appears to be a useful technique.

Doppler left ventricular (LV) inflow is reportedly affected by LV diastolic properties. We evaluated 48 subjects consisting of 27 patients with chronic mitral regurgitation (MR) and 21 patients with noncardiac disorders who received echocardiographic examinations. The deceleration rate divided by diastolic dimension (DR/Dd) derived from Doppler early diastolic LV inflow was correlated with the peak diastolic velocity divided by diastolic dimension (peak DV/Dd), a conventional index of LV diastolic function derived from the M-mode echocardiogram in the 48 patients, regardless of the presence of normal sinus rhythm or atrial fibrillation. LV diastolic function was then estimated by comparing perioperative echocardiographic examination and LV micro-and ultrastructural findings of biopsy specimens from 12 patients with MR who received mitral valve replacement. Fiber diameter, volume fraction of interstitial fibrosis (int. % Fib), and volume fractions of three intracellular components; the myofibrils (% MF), the sarcoplasmic reticulum (% SR) and the mitochondria (% MT), were measured in LV transmural biopsy specimens. DR/Dd was significantly correlated with peak DV/Dd before and after operation. Peak DV/Dd and DR/Dd were inversely correlated with int. % Fib and % SR, and were positively correlated with % MF. We subdivided the 12 MR patients according to their postoperative DR/Dd values as "recovered", and "non-recovered" based on their postoperative LV diastolic function. % MF was significantly lower in the 'non-recovered' group. Thus, DR/Dd can serve as an index of LV diastolic function. A decrease in % MF may inhibit the recovery of postoperative LV diastolic function.

Formalin-fixed, paraffin-embedded tissue sections of resected tumors from 90 patients were immunohistochemically studied to assess the prognostic value of proliferating cell nuclear antigen (PCNA) expression in non-small cell lung cancer. The individual tumors were classified into groups of high, moderate or low proliferative grade, and 38 (42.2%) patients had a high grade of proliferation. No statistically significant correlations were observed between PCNA grade and TNM status, pathological stage, resectability, histological type, degree of histological differentiation. Only vascular invasion significantly correlated with proliferative grade (p < 0.05). Survival analysis showed that patients with low proliferative grade tumors survived significantly longer (a 5-year survival rate of 83.3%) than those with high proliferative grade tumors (39.4%, p < 0.005). Cox's multivariate analysis revealed that PCNA grade was a significant prognostic determinant of survival. These results suggest that PCNA expression provides an independent prognostic variable for patients with non-small cell lung cancer and that it may be useful to consider this factor in treatment planning.

The relationship between MR configuration and pathological grade was studied in 41 histologically verified supratentorial astrocytic gliomas with a 0.5T superconductive MR system. The gliomas included 13 low-grade astrocytomas (LGAs), 14 anaplastic astrocytomas (AAs) and 14 glioblastoma multiformes (GBMs). MRI configurations were classified into nine criteria which were scored and statistically analyzed. The mean values of LGAs, AAs and GBMs were 0.45 +/- 0.31, 1.18 +/- 0.20 and 1.47 +/- 0.22. In each grade, MRI score increased as pathological grades increased (p < 0.01-0.001). LGAs had significantly lower values than AAs in five of the nine criteria (55.6%); heterogeneity, cyst or necrosis, edema or mass effect, border definition, and the degree of contrast enhancement, and lower values than GBMs in eight criteria (88.9%) except for hemorrhage. Three criteria (33.3%), heterogeneity, cyst or necrosis, and flow void sign were significantly higher in GBMs than AAs. The four variables, heterogeneity, cyst or necrosis, edema or mass effect and border definition, proved to be important factors related to the pathological grade in a multiple regression analysis.