Prognostic significance of L-type amino-acid transporter 1 expression in surgically resected pancreatic cancer.

BACKGROUND: The expression of L-type amino-acid transporter 1 (LAT1) is tumour-specific and has been shown to have essential roles in cell growth and survival. However, little is known regarding the clinical significance of LAT1 expression in pancreatic cancer. This study was conducted to determine the prognostic significance of LAT1 expression. METHODS: A total of 97 consecutive patients with surgically resected pathological stage I-IV pancreatic ductal adenocarcinoma were retrospectively reviewed. Tumour sections were stained by immunohistochemistry for LAT1, CD98, Ki-67 and vascular endothelial growth factor (VEGF), and microvessel density was determined by CD34 and p53. RESULTS: L-type amino-acid transporter 1 and CD98 were highly expressed in 52.6% (51/97) and 56.7% (55/97) of cases, respectively (P=0.568). The expression of LAT1 within pancreatic cancer cells was significantly associated with disease stage, tumour size, Ki-67, VEGF, CD34, p53 and CD98. L-type amino-acid transporter 1 expression was confirmed to be a significant prognostic factor for predicting poor outcome by multivariate analysis. CONCLUSION: L-type amino-acid transporter 1 expression is a promising pathological marker for the prediction of outcome in patients with pancreatic cancer.

Apical vesicles bearing inositol 1,4,5-trisphosphate receptors in the Ca2+ initiation site of ductal epithelium of submandibular gland.

In polarized epithelial cells, agonists trigger Ca2+ waves and oscillations. These patterns may be caused by the compartmentalization of inositol 1,4,5-trisphosphate (IP3)-sensitive Ca2+ pools into specific regions. We have investigated the relationship between the distribution of IP3 receptors (IP3Rs) and the spatiotemporal pattern of Ca2+ signaling in the duct cells of the rat submandibular gland (SMG). Using immunofluorescence, although labeling was somewhat heterogeneous, the IP3Rs were colocalized to the apical pole of the duct cells. Immunoelectron microscopy identified small apical vesicles bearing IP3R2 in some types of duct cells. Real-time confocal imaging of intact ducts demonstrated that, after carbachol stimulation, an initial Ca2+ spike occurred in the apical region. Subsequently, repetitive Ca2+ spikes spread from the apical to the middle cytoplasm. These apical Ca2+ initiation sites were found only in some "pioneer cells," rather than in all duct cells. We performed both Ca2+ imaging and immunofluorescence on the same ducts and detected the strongest immunosignals of IP3R2 in the Ca2+ initiation sites of the pioneer cells. The subcellular localization and expression level of IP3Rs correlated strongly with the spatiotemporal nature of the intracellular Ca2+ signal and distinct Ca2+ responses among the rat SMG duct cells.

Effects of retinoic acid on the activation and fusion of mouse alveolar macrophages induced by 1 alpha,25-dihydroxyvitamin D3.

1 alpha,25-dihydroxyvitamin D3 [1 alpha,25(OH)2D3] directly induces both fusion and cytotoxicity in murine alveolar macrophages. Unlike 1 alpha,25(OH)2D3, retinoic acid per se did not induce fusion of alveolar macrophages, but it greatly enhanced the 1 alpha,25(OH)2D3-induced fusion every time the macrophages were treated simultaneously with the two vitamins. The giant cells induced by the two vitamins were much larger than those induced by 1 alpha,25(OH)2D3 alone. The macrophages treated with 1 alpha,25(OH)2D3 started to fuse 36 h after the addition of the vitamin, whereas the macrophages pretreated with retinoic acid for 24 h began to fuse immediately after 1 alpha,25(OH)2D3 was added. 1 alpha,25(OH)2D3 and retinoic acid activated alveolar macrophages similarly, measured by the enhancement of glucose consumption and the induction of cytotoxicity against tumor cells, though 1 alpha,25(OH)2D3 was 100 times more potent than retinoic acid on a molar basis. Simultaneous treatment with physiological concentrations of 1 alpha,25(OH)2D3 (0.12 nM) and retinoic acid (10 nM) induced cytotoxicity additively. Morphological examinations revealed that the treated cells were enlarged and flattened with numerous filopodia. These results clearly indicate that both 1 alpha,25(OH)2D3 and retinoic acid similarly activate alveolar macrophages, and the activated state is prerequisite to the fusion of macrophages induced by 1 alpha,25(OH)2D3.

An ultrastructural study on the multinucleation process of mouse alveolar macrophages induced by 1 alpha,25-dihydroxyvitamin D3.

The multinucleation process of isolated alveolar macrophages induced by 1 alpha,25-dihydroxyvitamin D3 [1 alpha,25(OH)2D3] was examined using a scanning electron microscope (SEM) and a transmission electron microscope (TEM). At the beginning of culture, most of the macrophages were spherical in shape. During incubation with 1.2 X 10(-8) M 1 alpha,25(OH)2D3, spreading macrophages appeared among the spherical macrophages, and they increased in number. Spreading macrophages extended many cytoplasmic processes toward adjacent macrophages, and interdigitations of these processes between those of neighboring cells were often seen. Two types of cell contact have been observed in the 1 alpha,25(OH)2D3-treated cells. In some, cytoplasmic processes were put into the cytoplasm of the adjacent cells, where clathrinlike structures were observed at the inner membrane of the concave portion. In others, spreading macrophages occasionally came in contact with adjacent cells by a peripheral rim of their cytoplasm with gap junctions. Cytoplasmic continuity was rarely observed at the boundaries between the closely associated cells. The two types of cell contact were also found, though not frequently, in the untreated cells. These results indicate that 1 alpha,25(OH)2D3 promotes multinucleation of alveolar macrophages through spreading forms with the formation of gap junctions and the coated membrane invagination.

Exocytosis in living salivary glands: direct visualization by video-enhanced microscopy and confocal laser microscopy.

Although exocytosis is widely believed to involve granule movement, membrane fusion and the emptying of granule content, direct study of these processes has been difficult in living cells because of the limited resolution of conventional light microscopy. Using video-enhanced microscopy and confocal laser microscopy, we have now studied these processes in living rat parotid and submandibular gland acinar cells. Under a differential interference contrast (DIC) microscope equipped with a CCD camera and a high speed image processor, secretory granules were in general stationary even after secretory stimulation with isoproterenol (IPR). Following IPR stimulation, however, there were abrupt changes in light intensity of secretory granules, and many granules disappeared. Confocal microscopy was then performed to confirm whether the observed changes in granules were related to membrane fusion and content release. For this, cells were perfused with the fluid-phase tracer Lucifer Yellow; confocal images thus obtained clearly demonstrated the appearance of fluorescence in omega-shaped invaginations of the apical plasma membrane which corresponded to the sites at which changes were observed in DIC images. The time sequence analyses of confocal images showed that there was a repetitive appearance and disappearance of omega-shaped fluorescent foci at the apical plasma membrane until most of the granules were depleted. During this time, there did not appear to be any significant expansion of the apical plasma membrane and if endocytic uptake of the tracer occurred, it was below the limit of detection. These observations provide new insights into the exocytotic process in salivary glands and are at variance in some respects with previous interpretations made from electron microscopy.

Quantitative comparison of anti-fading mounting media for confocal laser scanning microscopy.

Fading is one of the major obstacles to reliable observation in fluorescence microscopy. Using a confocal laser scanning microscope (CLSM) coupled to a computer, we quantitatively measured fading of fluorescence to formulate an equation, evaluated the anti-fading ability of several anti-fading media, and restored the faded images to the original level according to this equation. NIH 3T3 cells were stained with fluorescein isothiocyanate (FITC)-phalloidin, mounted with several commercial and homemade anti-fade media, and observed with CLSM under repeated illumination. With any mounting medium, attenuation of fluorescence intensity at a certain pixel occurred stepwise and the decrease was proportional to the intensity of the previous scan. From these results, we formulated an equation that has three coefficients: anti-fading factor (A), indicating the ability to retard fading; fluorescent intensity at the first scan (EM(1)); and background fluorescence (B). The fluorescent intensity at a certain point following nth scan is given as EM(n) = EM(1) * A ((n-1)). This equation was available for restoring faded images to their original states, even after the image had faded to only 60% of its original intensity.

Cyclic nature of poly(3-hydroxyalkanoate) metabolism in Alcaligenes eutrophus.

The cyclic process of biosynthesis and degradation of poly(3-hydroxyalkanoate) (PHA) was studied in Alcaligenes eutrophus under conditions of nitrogen-limitation of growth. A. eutrophus cells, which had accumulated poly(3-hydroxybutyrate) (PHB) of 55 wt% content within cells from butyric acid, were transferred into a nitrogen-free medium containing pentanoic acid as the sole carbon source and cultivated at 30 degrees C and pH 7.5. The content of PHB in A. eutrophus cells decreased with time, whereas a copolyester of 3-hydroxybutyrate (HB) and 3-hydroxyvalerate (HV) units, P(HB-co-HV), was accumulated in the presence of pentanoic acid. Conversely, when A. eutrophus cells with 50 wt% content of P(HB-co-56% HV) were incubated in a nitrogen-free medium containing butyric acid, the content of P(HB-co-56% HV) decreased with time, whereas PHB was accumulated. These results indicate the cyclic nature of PHA metabolism in A. eutrophus under these conditions.

Confocal laser scanning microscopic and immunoelectron microscopic studies of the anatomical distribution of fibrillar IgA deposits in dermatitis herpetiformis.

BACKGROUND AND DESIGN: The fibrillar immunofluorescent pattern of IgA deposition in dermatitis herpetiformis is considered by most authorities to be a variant of the granular IgA pattern. It has been hypothesized that the fibrillar vs the granular pattern is related to longitudinal vs transverse sectioning of affected dermal microfibril bundles. However, direct evidence for this possibility has yet to be presented. Confocal laser scanning microscopy and immunoelectron microscopy were performed to determine the anatomical distribution of fibrillar IgA deposits, using skin specimens from a patient with typical fibrillar IgA deposition. OBSERVATIONS: Confocal laser scanning microscopy showed numerous fibrils stained with anti-IgA extending from the dermoepidermal junction to a depth of 50 to 110 microns in the dermis. They crossed each other at various angles to form a three-dimensional network. Immunoelectron microscopy demonstrated a diffuse dispersion of immune deposits on the surface of microfibrils of dermal microfibril bundles, with sporadic distribution of small aggregates, 0.1 to 0.3 micron in diameter. CONCLUSIONS: To our knowledge, this is the first article to present evidence for the actual distribution of fibrillar IgA. Insofar as the present case is concerned, the distribution of fibrillar IgA is greatly at variance with that indicated in previous reports on granular IgA. However, studies on more cases should be conducted to determine whether this is a distinctive feature of the fibrillar type of IgA deposition.

Biosynthesis and characterization of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) in Alcaligenes eutrophus.

Copolyesters of 3-hydroxybutyrate (3HB) and 4-hydroxybutyrate (4HB) were produced by Alcaligenes eutrophus at 30 degrees C in nitrogen-free culture solutions containing gamma-butyrolactone alone or with fructose or butyric acid as the carbon sources. When gamma-butyrolactone was used as the sole carbon source, the 4HB fraction in copolyester increased from 9 to 21 mol% as the concentration of gamma-butyrolactone in the culture solution increased from 10 to 25 g/l. The addition of fructose to the culture solution of gamma-butyrolactone resulted in a decrease in the 4HB fraction in copolyester. The copolyesters produced from gamma-butyrolactone and fructose by A. eutrophus were shown to have random sequence distribution of 3HB and 4HB units by analysis of the 125 MHz 13C n.m.r. spectra. In contrast, a mixture of random copolyesters with two different 4HB fractions was produced by A. eutrophus when gamma-butyrolactone and butyric acid were used as the carbon sources. These results are discussed on the basis of a proposed biosynthetic pathway of P(3HB-co-4HB). The copolyester films became soft with an increase in the 4HB fraction, and the elongation to break at 23 degrees C increased from 5 to 444% as the 4HB fraction increased from 0 to 16 mol%. The P(3HB-co-10% 4HB) film was shown to be biodegradable in an activated sludge.

High levels of GM(1)-ganglioside and GM(1)-ganglioside beta-galactosidase in the parotid gland: a new model for secretory mechanisms of the parotid gland.

A new model for the subcellular basis of parotid secretion is presented in this article. GM(1)-ganglioside, typically found in neural tissues, is shown to be abundant in the parotid gland. This ganglioside may play a central role in membrane turnover mechanisms underlying exocytosis/endocytosis in its role as a promoter of membrane fusion or a fusogen. The lysosome and lysosomal hydrolases also play a central role in this model in catabolism of GM(1)-ganglioside. Consequently, high levels of the lysosomal hydrolase acidic beta-galactosidase are demonstrated in the salivary gland. GM(1)-gangliosidosis of the parotid glands, as described in mice, appears to be the first single-gene heritable disease found so far in the salivary glands.

A case of rhabdomyosarcoma of the vagina in an elderly woman.

Most rhabdomyosarcomas of the vagina (RMSV) occur in infants and children up to six years old. RMSV in elderly patients is extremely rare. We report a case of a 70-year-old woman with RMSV. She had received surgery for uterine endometrial cancer one year before and a vaginal polypoid tumor was noted during routine follow-up vaginal examination. She was referred to our department for radiation therapy following partial tumorectomy of the lesion. She was given three sessions of intra-vaginal radiation therapy, once a week with 6 Gy at 7.5 mm below the vaginal surface and external irradiation of 50 Gy to the pelvis. However, paraaortal lymph node metastasis developed during initial radiation therapy. Furthermore, multiple bone metastases appeared at the completion of the radiation therapy. Six months after initial treatment the patient died from progression of the disease. Autopsy demonstrated small residual tumor at the primary site as well as multiple systemic metastases.

Dynamics of salivary secretion studied by confocal laser and scanning electron microscopy.

Plasma membrane events during secretion of the parotid and submandibular glands of humans and rats were observed in living cells by confocal microscopy and from the cytoplasmic side by scanning electron microscopy after removal of the cell organelles by the OsO4 maceration method. These new microscopic techniques revealed in acinar cells two distinct exocytosis-endocytosis coupling mechanisms elicited in response to different secretory stimuli. Beta-adrenergic stimulation with isoproterenol and its second messenger analog dibutyryl cyclic AMP evoked exocytosis after which fused granule membranes were individually removed from the luminal plasma membrane within several minutes. On the fused membrane area, small endocytotic vesicles about 100-150 nm in diameter were abundant. Muscarinic stimulation with carbachol also induced exocytosis, but in this case the fused granule membranes coalesced to form enlarged invaginations which stayed on the luminal membrane for more than 30 min. These invaginations, almost devoid of small endocytotic vesicles, were then pinched off from the luminal membrane and dispersed into the cytoplasm in the form of light microscopically detectable vesicles. After treatment with isoproterenol and carbachol, acinar cells exhibited different distributional changes of F-actin around the fused membrane area, suggesting the involvement of microfilaments in regulating the membrane events following exocytosis.

Relationship between amylase and fluid secretion in the isolated perfused whole parotid gland of the rat.

Whole gland perfusion technique was applied to rat parotid glands to assess whether amylase affects fluid secretion. Control perfusion without any secretagogue evoked no spontaneous secretion. Carbachol (CCh 1 microM) induced both amylase and fluid secretion with distinctive kinetics. Fluid secretion occurred constantly at 40-120 microliter/g-min (average plateau was 60 microliter/g-min), whereas amylase secretion exhibited an initial peak (10 mg maltose/30 s per g wet w. of the gland), followed by a rapid decrease to reach a plateau level of 1 mg maltose/30 s later than 1.5-2 min. Isoproterenol (Isop 1 microM) alone did not induce fluid secretion although it evoked amylase secretion as measured in isolated perfused acini. Addition of Isop during CCh stimulation evoked a rapid and large rise in amylase secretion to 15 mg maltose/30 s accompanied by the increase in oxygen consumption. However, the fluid secretion exhibited a rather gradual decrease. These findings suggest that control of salivary fluid secretion is independent of the amylase secretion system induced by CCh and/or Isop. Morphological observations carried out by HR SEM and TEM revealed exocytotic profiles following Isop stimulation. CCh stimulation alone seldom showed -exocytotic profiles, suggesting a low incidence of amylase secretion during copious fluid secretion. Combined stimulation of CCh and Isop induced both vacuolation and exocytosis along intercellular canaliculi. During washout of secretagogues, lysosomal digestion of excess membrane took place.

Human salivary gland parenchymal cells seen by SEM from the cytoplasmic side using a new osmium maceration method.

By removing all or most organelles, we have exposed the cytoplasmic side of the plasmalemma and its specializations in serous cells and in cells of striated and excretory ducts of human major salivary glands. The areas of plasmalemma located beneath the lumen and those bordering the intercellular canaliculi are covered by evenly distributed particles arranged in a continuous band and, below it, in regularly spaced clusters. A similar pattern of particles is seen on the internal aspects of the juxtaluminal plasmalemma of cells of both striated and excretory ducts. Small isolated clusters of particles are seen in other regions of serous and ductal cells as well, being particularly numerous along the basal processes of cells of striated ducts. A distribution of particles resembling that present along intercellular canaliculi of serous cells also is seen on the plasmalemma bordering the biliary canaliculi where, however, the clusters look smaller and farther apart. Large clusters of particles, matching those seen on salivary glands and on liver, are present at the base of the short processes of cells of the stratum spinosum of squamous stratified epithelia. Since the sites of location of the clusters closely correspond to the areas where transmission electron microscopy (TEM) reveals the presence of desmosomes, we believe that the clusters may be related to these cellular junctions. Of more difficult interpretation are the particles present on the juxtaluminal band corresponding both to the zonula occludens and to the zonula adhaerens.

Cell biology of human salivary secretion.

We treated surgical specimens of human parotid and submandibular glands in vitro to manipulate the receptor-signaling cascade pharmacologically and analyzed cellular responses by light microscopy on epoxy embedded sections. Treatment of specimens with the b-agonist, isoproterenol, and with the second messenger analog, dibutyryl cyclic AMP, stimulated serous acinar cells to engage in exocytosis and degranulation. The muscarinic agonist, carbachol, and the calcium ionophore, A23187, on the other hand, elicited formation of "vacuoles" in the cytoplasm of serous acinar cells. Taking previous in vivo human and animal studies into account, these changes are suggested as the morphological expression of enzyme release and fluid secretion, respectively. Specimens obtained from patients over 70 years old exhibited poor response even though their morphological appearance remained intact. Aged salivary glands are thus suggested to experience a decline in their secretory activity at the cellular level, probably by impairment of the signaling processes downstream to the receptor activation and second messenger production.

Cytoskeletal regulation of human salivary secretion studied by high resolution electron microscopy and confocal laser microscopy.

To study the cell regulation mechanisms of human salivary secretion, surgical specimens of human parotid and submandibular glands were treated in vitro with isoproterenol (beta-agonist), carbachol (muscarinic agonist), and cytochalasin D (microfilament disruptive agent), and morphological changes occurring in serous acinar cells were observed. Control acinar cells treated without secretagogues exhibited only occasional examples of exocytosis. Microfilaments, revealed by transmission electron microscopy (TEM) and confocal laser scanning microscopy (CLSM) of F-actin fluorescence stained by rhodamine-phalloidin, were localized underneath the luminal membrane to separate the secretory granules from the luminal membrane. Following isoproterenol treatment, secretory granules made direct contact with the luminal membrane and many omega-shaped exocytotic profiles appeared. TEM and scanning electron microscopy (SEM) showed these profiles to be of granule size or somewhat smaller and to be provided on their cytoplasmic surface with coated pits. Furthermore, CLSM detected the appearance of F-actin fluorescence around the exocytosed granule membranes. Carbachol treatment also evoked the formation in acinar cells of omega-shaped exocytotic profiles some of which were larger than the granules and which exhibited neither coated pits nor associated F-actin fluorescence. To determine if microfilaments regulate the post-exocytotic process of membrane retrieval, we combined isoproterenol treatments with cytochalasin D or carbachol. Following these treatments, F-actin fluorescence surrounding the exocytosed membrane was dispersed or diffused and the exocytotic profiles enlarged remarkably. These results led to the hypothesis that exo/endocytotic processes in human salivary serous acinar cells are regulated differently under autonomic receptor control mediated by microfilaments.

[The treatment of staghorn calculi].

Between May 1989 and November 1991, 19 staghorn calculi were treated by extracorporeal shock-wave lithotripsy (ESWL) with a Dornier MFL 5000 or Northgate SD-3. The 19 calculi were evaluated. Treatment was with monotherapy by ESWL in 9, combination percutaneous nephrolithotomy (PNL)-ESWL in 9, and nephrostomy in 1. Of the patients, 14 had a cross stent catheter pre-ESWL treatment to improve fragment evacuation. Radiologic follow up in 19 kidneys revealed that 57.9% were stone free. We arbitrarily separated our cases into 3 groups: struvite renal calculi, calcium carbonate calculi and others. Result of stone-free rate was 100% for stones consisting of struvite, and 14. 3% for stones consisting of calcium carbonate. In our opinion, the best indication of monotherapy by ESWL is for staghorn calculi, which consists of struvite, without marked dilation of pelvis and calyces.

[Spindle cell carcinoma of the urinary bladder: a case report].

A 74-year-old man with spindle cell carcinoma of the urinary bladder is reported. He presented to our clinic with gross hematuria resulting in bladder tamponade. Transurethral resection (TUR) was performed in order to control severe hematuria intractable with conservative therapy. Histologically the tumor invaded the muscle layer and was composed of two components, small foci of transitional cell carcinoma and numerous spindle cells with severe atypia and then the transition was recognized between them. Immunohistochemically transitional cell carcinoma was intensely positive for cytokeratin (CK) and epithelial membrane antigen (EMA), but negative for vimentin (VIM). Moreover some parts of spindle cells were weakly stained for CK and EMA. Three months after TUR, multiple pulmonary metastases and moderate right hydronephrosis occurred and he died of respiratory insufficiency one month later.

[Clinical evaluation of cefmetazole in urological infections].

Clinical efficacy of Cefmetazole was evaluated at four university hospitals and their related hospitals in Nagoya. For the treatment of urinary tract infections with or without complications, 177 patients were administered Cefmetazole. Of these patients, 69 had chronic complicated urinary tract infection defined in the UTI manual and 20 had simple acute pyelonephritis. The other urological infections for which Cefmetazole was administered included prostatitis, epididymitis, urosepsis and wound infections. Fifty four patients were given Cefmetazole intravenously after urological operation to prevent wound and urinary tract infections. The overall clinical efficacy of Cefmetazole for UTI was 76.8%; 84.4% for group 1, 85.7% for group 3, 75% for group 4, 44.4% for group 5 and 66.6% for group 6. In acute pyelonephritis due to E. coli, Klebsiella, Serratia, S. aureus, alpha-Streptococcus and S. epidermidis all patients were cured by Cefmetazole administration. Clinical efficacy of Cefmetazole was assessed to be excellent in 6 cases of prostatitis and 6 cases of acute epididymitis. E. Coli, Serratia and some organisms disappeared from blood after the administration of Cefmetazole but Pseudomonas persisted even after treatment. Postoperative administration of Cefmetazole was effective for eradication of bacteria from the urine in 26 out of 30 patients and in prevention of infection in 24 cases. After the administration of Cefmetazole skin eruption was observed in one patient and nausea in another. Slight elevation of GOT, GPT and total bilirubin was noted in 3 of the 177 patients after medication.

[Clinical study of renal cell carcinoma].

Previously, we reported the effects of human lymphoblastoid interferon (HLBI), using a transplantable human renal cell carcinoma strain (AM-RC-3) in nude mice established in our laboratory. An overall anticancer effect was found from its combination with UFT (Ft-207t uracil). In the present investigation, we examined the clinical effectiveness when HLBI was administered alone or in combination with UFT to the patients. Seventy-three patients who had undergone curative surgery were divided into 3 groups, according to the type of adjuvant therapy. The HLBI group consisted of 38 patients, including those administered the agent alone over 50 times for more than six months, and or those to whom it was given in combination with UFT. The second group of 23 patients had been treated with hormones, radiotherapy, or with an anticancer drug (Fluoride pyrimidine group), while the last group of 17 patients underwent no postoperative adjuvant therapy. The survival rate calculated by the Kaplan-Meier method revealed no significant effects of HLBI on the survival period, but a significant difference (p < 0.05) was found in the HLBI group compared to the other groups in terms of much higher non-recurrence rate. When HLBI was administered alone or in combination with UFT, a definite anticancer effect was seen in 6 (complete response 3, partial response 2, minor response (MR) 1, no change 5, progression of disease 14) of the 25 treated patients. Fourteen of the 25, treated patients had postoperative recurrence, and 11 patients had distant metastases, at the time of diagnosis which were considered to be progressive and measurable lesions. In 6 patients the response was better than MR, with the effective rate being 24%. Four of the 6 patients had received HLBI in combination with UFT, which suggests a clinical effect in this combination. However, the effectiveness was limited to the lung lesions, more effective treatment of the lesions in other sites is required.