Capturing drug responses by quantitative promoter activity profiling.

Quantitative analysis of cellular responses to drugs is of major interest in pharmaceutical research. Microarray technologies have been widely used for monitoring genome-wide expression changes. However, this approach has several limitations in terms of coverage of targeted RNAs, sensitivity, and quantitativeness, which are crucial for accurate monitoring of cellular responses. In this article, we report an application of genome-wide and quantitative profiling of cellular responses to drugs. We monitored promoter activities in MCF-7 cells by Cap Analysis of Gene Expression using a single-molecule sequencer. We identified a distinct set of promoters affected even by subtle inhibition of the Ras-ERK and phosphatidylinositol-3-kinase-Akt signal-transduction pathways. Furthermore, we succeeded in explaining the majority of promoter responses to inhibition of the upstream epidermal growth factor receptor kinase quantitatively based on the promoter profiles upon inhibition of the two individual downstream signaling pathways. Our results demonstrate unexplored utility of highly quantitative promoter activity profiling in drug research.CPT: Pharmacometrics & Systems Pharmacology (2013) 2, e77; doi:10.1038/psp.2013.53; published online 25 September 2013.

Beneficial effect of interferon-beta treatment in patients with multiple sclerosis is associated with transient increase in serum IL-6 level in response to interferon-beta injection.

In order to predict the clinical benefit of interferon-beta (IFN-beta) to patients with multiple sclerosis (MS), the following markers were investigated; (1) chronological change of cytokines (IFN-gamma, TNF-alpha, IL-6, IL-10, and TGF-beta) after administration of IFN-beta, (2) untoward effects of IFN-beta such as headache and arthralgia, (3) backgrounds of the patients such as age and relapse rate, (4) efficacy of IFN-beta therapy assessed by the change of relapse rate and progression of disability. Chronological blood sampling was performed 0, 10, and 24 h after injection of IFN-beta. The increase of serum IL-6 level in response to IFN-beta administration was associated with headache, arthralgia, relapse rate before treatment, and disability score at the initiation of the therapy. Significant association of change of serum TNF-alpha with age and headache was also observed. The important finding in this study was that patients with a transient increase in IL-6 in response to IFN-beta showed a slow disease progression. This result suggests that this transient increase in the serum IL-6 predicts favorable response to IFN-beta treatment.

Altered expression of beta-catenin without genetic mutation in intrahepatic cholangiocarcinoma.

beta-catenin which has a role in E-cadherin mediated cell-to-cell adhesion, and is also involved in Wnt signaling pathways as a downstream signaling molecule accumulating in the cytoplasm and nucleus constitutively activates Tcf/LEF-associated transcription of oncogenic genes. We examined the expression pattern and the genetic alteration of beta-catenin to determine the role of beta-catenin in cancer formation and/or progression in intrahepatic cholangiocarcinoma (ICC). beta-catenin expression was immunohistochemically examined in 71 surgically resected ICC samples, and correlation between the expression pattern and clinicopathologic factors was investigated. Mutation analysis of beta-catenin exon 3, which included the responsible element for Wnt signaling was done in 55 samples, using PCR-SSCP and direct sequence methods. Immunohistochemical analysis revealed the reduced membranous expression of beta-catenin in 58 (82%) ICCs and aberrant nuclear expression in 11 (15%) ICCs. The membranous expression was preserved in 62% of the papillary adenocarcinomas, and was frequently reduced in tumors with a poorer histological differentiation (84%), with a significant difference (P =.01). Genetic analysis showed that none of the 55 ICCs examined carried mutations in beta-catenin exon 3. The present study indicates that reduced membranous expression of beta-catenin is associated with non-papillary ICCs which have a more malignant behavior, and that nuclear translocation of beta-catenin results in oncogenic events. Mutations in beta-catenin exon 3 do not appear to be responsible for nuclear translocation of beta-catenin in ICCs.

The role of overexpression and gene amplification of cyclin D1 in intrahepatic cholangiocarcinoma.

BACKGROUND/AIMS: Intrahepatic cholangiocarcinoma (ICC) is a primary liver malignant tumor with an extremely poor prognosis, but less attention has been directed to factors related to molecular carcinogenesis, including cell cycle proteins. We examined the expression and gene amplification of cyclin D1, the cell cycle regulating protein. Our objective was to evaluate correlations with clinicopathological factors in ICC. METHODS: Cyclin D1 overexpression and cellular proliferative activity (Ki-67 labeling index) were investigated immunohistochemically, and 20 cases were further investigated for cyclin D1 gene amplification, using differential PCR. We examined the correlation between the expression and gene amplification of cyclin D1 and clinicopathological factors, including overall survival in patients with ICC. RESULTS: Immunohistochemical analysis revealed an overexpression of cyclin D1 protein in 28 of 66 subjects with ICCs (42%). The cyclin D1 overexpression was associated with poor histological differentiation (P = 0.04), high cellular proliferative activity (P < 0.01), and a poor prognosis (P = 0.02) by univariate analysis, although it is not an independent prognostic factor by multivariate analysis. Cyclin D1 gene amplification was confirmed in five of the 20 patients. Of those five cases of ICC, all had poor histological differentiation, and four of the five ICCs (80%) showed evidence of cyclin D1 immunoreactivity. CONCLUSIONS: Overexpression and gene amplification of cyclin D1 are frequent and contribute to dedifferentiation and cellular proliferative activity of ICCs, and overexpression also indicates a poor prognosis for patients with ICC.

Clinicopathological characteristics of patients with extrahepatic recurrence following a hepatectomy for hepatocellular carcinoma.

BACKGROUND/AIMS: None of the previous studies have compared the prognosis or clinicopathological factors between the patients with extrahepatic recurrence and those with intrahepatic recurrence of hepatocellular carcinoma after a hepatic resection. METHODOLOGY: The clinicopathological features and prognoses of patients with extrahepatic recurrence after a curative hepatectomy for hepatocellular carcinoma were investigated. RESULTS: Twenty-three patients with extrahepatic recurrence had more advanced-stage hepatocellular carcinoma at the primary operation compared to 186 patients with intrahepatic recurrence. After adjusting for tumor size, the prognosis of the 2 groups were comparable. However, among the patients with hepatocellular carcinoma exceeding 5 cm in diameter, the number of patients whose plasma levels of des-gamma-carboxy prothrombin was higher than 2.0 AU/mL in the patients with extrahepatic recurrence (62.5%) was significantly more (P < 0.05) than that in the patients with intrahepatic recurrence (20.0%). On the other hand, the prognosis of the 13 patients with extrahepatic recurrence alone was significantly better than in the 10 patients with both intrahepatic and extrahepatic recurrences. The prognoses of the 3 patients who underwent a surgical resection for isolated extrahepatic recurrence were markedly better than that of the remaining 10 patients only treated palliatively. CONCLUSIONS: If patients have tumors exceeding 5 cm in diameter and their plasma levels of des-gamma-carboxy prothrombin are higher than 2.0 AU/mL, more careful follow-up examinations than usual may thus be necessary in order to detect extrahepatic recurrence as early as possible. Furthermore, a surgical resection for the isolated extrahepatic recurrence of hepatocellular carcinoma is also recommended to produce long-term survivors.

Lymphocytic hypophysitis: its expanding features.

Lymphocytic hypophysitis is classically defined as an inflammatory disorder confined to adenohypophysis. However, it has recently been indicated that infundibuloneurohypophysitis underlies some subsets of central diabetes insipidus (DI). Therefore, lymphocytic hypophysitis can be considered a syndrome including disorders of both the anterior pituitary (lymphocytic adenohypophysitis) and the posterior pituitary (lymphocytic infundibuloneurohypophysitis). We describe a 77-yr-old woman with lymphocytic hypophysitis presenting with headache, diplopia, general malaise and appetite loss. Head magnetic resonance imaging (MRI) demonstrated pituitary swelling and dura mater thickening on the dorsum sella. Endocrinological investigations revealed both anterior and posterior pituitary dysfunction associated with primary hypothyroidism due to Hashimoto's thyroiditis. Headache and diplopia spontaneously disappeared, and anterior pituitary dysfunction, general malaise and appetite loss improved after taking 10 mg hydrocortisone daily, although ACTH hyposecretion persisted. Pituitary swelling was thereafter reduced but the dura mater thickening persisted. We suggest that this case may represent a variant of lymphocytic hypophysitis in which chronic inflammatory process involves both the anterior and the posterior pituitary gland, infundibulum, dura mater on the dorsum sella and cavernous sinus. Regarding expanding features of lymphocytic hypophysitis, it may be considered a syndrome including heterogeneous disorders, of which the pathogenesis remains to be elucidated.

Highly selective one-pot synthesis of spirophosphoranes exhibiting reversed apicophilicity by oxidation of dianions generated from P-H spirophosphorane.

[see reaction]. Mild and highly selective one-pot procedures for obtaining phosphoranes that exhibit reversed (O-cis) apicophilicity are described. On the basis of the procedures, O-cis phosphorane bearing an aryl group (R = 2,4,6-tri-i-propylphenyl) could be isolated for the first time; the procedure is also applicable for alkyl derivatives. Particularly effective was the use of I2 as an oxidizing reagent.

The role of p27kip1 protein expression on the biological behavior of intrahepatic cholangiocarcinoma.

Recent studies have reported that a decreased p27kip1 protein (p27) expression is associated with aggressive tumor behavior in several human malignancies. In this study, we examined the role of p27 expression in intrahepatic cholangiocarcinoma (ICC) and the noncancerous intrahepatic bile duct epithelium and assessed its association with clinicopathologic features, especially the influence of decreased p27 expression by the tumor on its prognosis as determined by immunohistochemistry. Almost all noncancerous bile duct epithelia show positive nuclear staining and reveal a faintly positive reaction in the cytoplasm. In 62 surgically resected ICCs, a variable degree of positive p27 nuclear staining is recognized. A subset (13 of 62 cases; 21.0%) of the carcinomas displayed cytoplasmic staining. We classified 62 cases into 2 groups, a low expression group (< 50% of p27 nuclear positive cells) and a high expression group (> or = 50% of p27 nuclear positive cells), and tested for the association between positive expression and clinicopathologic features. We found that a low expression of nuclear p27 was correlated with positive vascular invasion (P < .05). The survival curve of the group with low p27 nuclear expression was significantly lower than that of the group with high p27 nuclear expression (P < .005). According to the multivariate analyses, low p27 expression can be considered an independent prognostic factor whose occurrence indicates a worse prognosis. We conclude that a decreased p27 expression adds an independent parameter that can be used when determining the prognosis of patients with ICC.

Evaluation of the modified maxillary protractor applied to Class III malocclusion with retruded maxilla in early mixed dentition.

The purpose of this study was to evaluate the effects of orthodontic treatment with a maxillary protraction bow appliance on anterior crossbite patients with Class III malocclusion in the mixed dentition. The 29 patients treated with a maxillary protraction bow appliance (11 boys, 18 girls) were compared with 25 matched, untreated controls with anterior crossbite (10 boys, 15 girls). The mean age before treatment was 8 years 7 months (range, 6 years 3 months to 11 years 6 months). The mean treatment period to achieve a normal overjet was 10.2 months (range, 5 to 18 months). Fifty-nine cephalometric angular and linear parameters were compared between the treated group and the untreated controls using the analysis of variance and the paired t test to evaluate the effect of gender and the maxillary protraction bow appliance treatment. Skeletal and dentoalveolar advancement of the maxilla and retrusion of the mandible contributed significantly to the improvement of Class III malocclusion in the treated group. These results suggest that a maxillary protraction bow appliance is effective for correcting anterior crossbite with a retruded maxilla in the early mixed dentition.

Characteristics of sarcomatous cholangiocarcinoma of the liver.

BACKGROUND/AIMS: The aim of this study was to clarify the clinicopathological characteristics of intrahepatic cholangiocarcinoma with sarcomatous changes. METHODOLOGY: Four cases of cholangiocarcinoma with sarcomatous change were identified and investigated. The clinicopathological findings, including the results of immunohistochemical staining, were investigated in comparison with those of ordinary cholangiocarcinoma. RESULTS: Two of them exhibited pyrexia as the initial symptom. The serum alkaline phosphatase level in sarcomatous cholangiocarcinoma was significantly lower than that in ordinary cholangiocarcinoma. Both the serum carcinoembryonic antigen and carbohydrate 19-9 level in sarcomatous cholangiocarcinoma also tended to be lower than those in ordinary cholangiocarcinoma. The carcinomatous component of all tumors was mostly poorly differentiated adenocarcinoma. The associated microscopic findings were as follows: lymphocyte infiltration in the tumor, accompanied by both necrosis and extensive lymph node metastases. Three of them predominantly exhibited spindle-shaped sarcomatous changes, whereas the other case predominantly demonstrated pleomorphic-type sarcomatous changes. The sarcomatous area was positive for both vimentin, a mesenchymal marker, and for epithelial markers, furthermore, in 3 of 4 cases, the carcinoma portions were also positive for vimentin. The overall survival curves were not significantly different between the 2 groups, however, no long-term survivor was found in sarcomatous cholangiocarcinoma. CONCLUSIONS: The main characteristics of cholangiocarcinoma with sarcomatous changes are considered to be as follows: 1) often demonstrating pyrexia as a symptom; 2) not always demonstrating remarkable abnormal findings in the laboratory data including tumor markers; 3) histologically showing poorly differentiated adenocarcinoma; and 4) showing a very poor prognosis (especially, in a pleomorphic-type).

Small hepatocellular carcinoma with minute satellite nodules.

BACKGROUND/AIMS: To investigate the clinicopathologic characteristics of small hepatocellular carcinoma with minute satellite nodules. METHODOLOGY: We investigated the clinicopathologic characteristics of 131 solitary small (< or = 2.0 cm in diameter) hepatocellular carcinomas including 105 hepatocellular carcinomas without minute satellite nodules and 17 hepatocellular carcinomas with minute satellite nodules smaller than 5 mm, and also discuss the clinical significance. RESULTS: None of the clinical backgrounds of the patients and pathologic features of the main tumor, except for the average of preoperative serum alpha-fetoprotein, were significantly different between the two groups. Firstly, minute satellite nodules demonstrated that the maximum diameter of all minute satellite nodules was 1.5-4.0 mm, secondly, the moderately to poorly differentiated hepatocellular carcinomas had 4 or more minute satellite nodules within 1 cm from the main tumor, while well differentiated hepatocellular carcinomas may have 1 or 2 minute satellite nodules 6 cm or more away, and thirdly, 4 or more minute satellite nodules may present within 1 cm in intrahepatic metastasis cases, while 1 or 2 minute satellite nodules may be present 6 cm or more away from the main tumor in multicentric occurrence cases. CONCLUSIONS: At least 13% of solitary small hepatocellular carcinomas had preoperatively undetectable minute satellite nodules. In case of moderately to poorly differentiated hepatocellular carcinomas, hepatic resection as well as percutaneous ethanol injection should be performed including the surrounding liver tissue at least 1.0 cm from the main nodule. On the other hand, in well-differentiated hepatocellular carcinomas, which may indicate multicentric occurrence, closer observation and careful follow-up after therapy are recommended.

A resected case of combined hepatocellular carcinoma and cholangiocarcinoma associated with cystic formation.

Combined hepatocellular carcinoma and cholangiocarcinoma is a rare tumor. In addition, both hepatocellular carcinoma and cholangiocarcinoma are rarely associated with cystic lesions. We herein present a 62-year-old Japanese woman with combined hepatocellular carcinoma and cholangiocarcinoma which was associated with a rapidly enlarging cystic lesion. Both abdominal ultrasonography and computed tomography revealed a cyst with a solid portion in the left hepatic lobe. A partial hepatectomy was performed on the basis of a tentative diagnosis of a cystadenocarcinoma of the liver, while the diagnosis based on immunohistochemical studies was combined hepatocellular carcinoma and cholangiocarcinoma with cystic formation. The patient died of tumor recurrence, such as intrahepatic metastases and extensive lymph node metastases, 6 months after the operation. The prognosis of this entity, which has never been reported in the English medical literature and is difficult to preoperatively differentiate from hepatic cystadenocarcinoma, therefore seems to be extremely poor.

Esthetic management of an unerupted maxillary central incisor with a closed eruption technique.

In cases of orthodontic traction of unerupted teeth, gingival recession and long clinical crowns are often seen. Satisfactory esthetic demands are not always met. A case of an unerupted maxillary central incisor is presented, in which traction was used, with emphasis on the surgical technique and the direction of the pull. The result was clinically favorable. The appearance of gingival margin was restored.

Clinicopathological characteristics of surgically resected minute hepatocellular carcinomas.

BACKGROUND/AIMS: The multistep development of overt hepatocellular carcinoma from very well-differentiated early hepatocellular carcinoma, and of early hepatocellular carcinoma from adenomatous hyperplasia has been strongly suggested. The clinicopathologic and immunohistochemical characteristics of solitary minute hepatocellular carcinomas smaller than 1 cm in size have yet to be clarified. METHODOLOGY: Fourteen minute hepatocellular carcinomas were divided into 2 groups consisting of: 1) hepatocellular carcinoma of hepatitis B surface antigen positive patients (B-HCC) (n = 5), and 2) hepatocellular carcinoma of hepatitis C virus antibody positive patients (C-HCC) (n = 9), then they were all analyzed histopathologically and clinicopathologically. Immunohistochemical studies were also performed using the antibodies against p53 protein. RESULTS: Six of the 14 minute hepatocellular carcinoma were demonstrated to be moderately or poorly differentiated tumors. Among the 8 well-differentiated minute hepatocellular carcinomas, 2 tumors already contained less differentiated components. B-HCC tended to be less differentiated than C-HCC (P < 0.05). Adenomatous hyperplasia was detected in only 2 cases of C-HCC. Small cell liver dysplasia was detected significantly more frequently in C-HCC than in B-HCC (P < 0.05). The prognosis of the 14 minute hepatocellular carcinomas varied considerably. Immunohistochemically, some tumor cells were positive for p53 in 3 cases. CONCLUSIONS: Our study suggests that 1) the multistep carcinogenesis through adenomatous hyperplasia may not be so frequent, 2) De novo carcinogenesis from not only well-differentiated hepatocellular carcinoma, but also from less differentiated hepatocellular carcinoma, especially B-HCC, may be present, 3) the carcinogenesis in the B-HCC cases may behave differently from that in C-HCC cases, and 4) minute hepatocellular carcinomas demonstrate varying prognoses after hepatectomy.

Expression of matrix metalloproteinase-9 in surgically resected intrahepatic cholangiocarcinoma.

BACKGROUND: Matrix metalloproteinase-9 (MMP-9) has recently been reported to be related to cancer cell invasion. This study was performed to clarify the expression of MMP-9 in surgically resected intrahepatic cholangiocarcinoma (IHCC). METHODS: In 37 patients with IHCC who underwent a surgical resection, the expression of MMP-9 and the clinicopathologic characteristics of MMP-9-positive IHCC were investigated. The expression of MMP-9 was immunohistochemically detected in 16 (43%) of 37 IHCC. The patients were divided into MMP-9 (-) IHCC (n = 21), MMP-9 (+) IHCC (n = 12), and MMP-9 (++) IHCC (n = 4). RESULTS: The survival rate after surgical resection in patients with MMP-9 (-), (+), and (++) IHCC, was 66%, 39%, and 0% at one year, 50%, 32% and 0% at 3 years, respectively (P = .001). The incidence rate of lymph node metastasis was 6 (28%) of 21 in MMP-9 (-) patients, 7 (58%) of 12 in MMP-9 (+) patients and 4 (100%) of 4 in MMP-9 (++) patients. The incidence rate of lymph node metastasis increased in proportion to an increase in the expression of MMP-9 in IHCC (P = .02). Recurrence in the lymph node was more common in patients with MMP-9 (+) and (++) cancers than in those with MMP-9 (-) cancers. CONCLUSIONS: The expression of MMP-9 in IHCC was a prognostic factor related to lymph node metastasis.

Vascular changes in hepatocellular carcinoma: correlation of radiologic and pathologic findings.

OBJECTIVE: Our objective was to analyze the hemodynamic properties and vascular supply changes in the carcinogenesis of hepatocellular carcinoma. MATERIALS AND METHODS: Ten nodules (nine patients) (one early, three early-advanced, and six advanced cases of hepatocellular carcinoma) less than 3 cm in diameter were selected from 45 patients (50 nodules) who underwent CT arteriography and CT during arterial portography. These images were correlated with histopathologic findings. Ratios of all microscopically counted (normal hepatic and abnormal) arteries, normal hepatic arteries, and portal veins in each nodule to those in the surrounding liver were calculated. RESULTS: Early hepatocellular carcinoma (one early case and early areas in three early-advanced cases) had low attenuation on CT arteriography and isoattenuation on CT during arterial portography. Advanced hepatocellular carcinoma (six advanced cases and advanced areas in three early-advanced cases) had high attenuation on CT arteriography and low attenuation on CT during arterial portography. In early hepatocellular carcinoma, the ratios of all arteries, normal hepatic arteries, and portal veins were 1.21 +/- 0.07, 0.60 +/- 0.07, and 0.73 +/- 0.06, respectively. In advanced hepatocellular carcinoma, the ratios were 2.66 +/- 0.26, 0.08 +/- 0.04, and 0.07 +/- 0.03, respectively. CONCLUSION: In early hepatocellular carcinoma, the combination of normal hepatic artery degeneration and preserved portal veins results in low attenuation on CT arteriography and isoattenuation on CT during arterial portography. In advanced hepatocellular carcinoma, the combination of neoplastic (abnormal) arterial development by angiogenesis and obliteration of portal veins results in high attenuation on CT arteriography and low attenuation on CT during arterial portography. These findings are a characteristic difference between early and advanced hepatocellular carcinoma.

The significance of stromal desmoplasia in intrahepatic cholangiocarcinoma: a special reference of 'scirrhous-type' and 'nonscirrhous-type' growth.

Fifty-eight surgically resected cases of intrahepatic cholangiocarcinoma (ICC) were examined grossly and microscopically. We microscopically categorized the ICC into two groups, "scirrhous-type" ICC (SICC) and "nonscirrhous-type" ICC (NSICC), according to the degree of the stromal desmoplasia in the tumor. We next assessed the histologic characteristics including proliferative activities and prognoses. Additionally, the prognostic factors of ICC were examined by univariate and multivariate analyses. Histologically, the incidences of lymphatic permeation and perineural invasion in SICC (85.7% and 100.0%, respectively) were significantly higher than those in NSICC (47.7% and 45.6%, respectively). The mean MIB-1 labeling index of SICC (mean+/-standard deviation, 29.9%+/-14.3%) was significantly higher than that of NSICC (mean+/-standard deviation, 19.1%+/-14.3%). Additionally, the 1- and 3-year survival rates of the SICC were 64.3% and 22.0%, respectively, and those of NSICC were 72.8% and 55.8%, respectively. The survival rate of SICC was significantly lower than that of NSICC. Overall, lymphatic permeation, intrahepatic metastasis, and lymph node metastasis were all considered to be independent prognostic factors for a worse prognosis based on the multivariate analyses, whereas our new category based on stromal desmoplasia or other variables were not. Our study thus indicates that lymphatic permeation, perineural invasion, and a higher proliferative activity of tumor cells carry a poorer prognosis in SICCs than in NSICCs. Although the overall independent prognostic factors were lymphatic permeation, intrahepatic permeation, and lymph node metastasis based on a multivariate analysis, a detailed histologic examination related to the stromal desmoplasia in the tumor is also considered to be useful for evaluating the biologic behavior of the ICC.

Cell proliferation and cell loss in nodule-in-nodule hepatocellular carcinoma.

BACKGROUND/AIMS: In order to clarify the significance of the balance between cell proliferation and cell loss during the progression of hepatocellular carcinoma, 16 operative specimens of nodule-in-nodule hepatocellular carcinoma were investigated. METHODOLOGY: In 16 specimens, cell proliferation was evaluated by the expression of Ki-67 nuclear antigen, and cell loss was also examined by the method of terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL). The expressions of p53 protein, bcl-2 protein and Fas antigen were also investigated to clarify the relationship between their expression and cell kinetics. RESULTS: The Ki-67 labeling index of the inner nodules was higher than that for the outer nodules (18.9% vs. 7.2%; p < 0.05) and the TUNEL labeling index of the inner nodules was also higher than that for the outer nodules (12.8% vs. 6.6%; p < 0.05). The increasing rate of the Ki-67 labeling index from Edmondson's grade I to II was 3.9 +/- 3.0, that from grade II to III was 3.9 +/- 2.4, while the increasing rate of the TUNEL labeling index from grade I to II was 2.7 +/- 0.3 and that from grade II to III was 1.7 +/- 0.2 (p < 0.05). p53 Protein was observed in 5 cases, while bcl-2 protein was found in 4 cases in the border area of the inner nodule. However, Fas antigen was found in none of the examined cases. Regarding the Ki-67 positive rate in the inner nodule, the Ki-67 positive rate in the p53 protein positive cases was significantly higher than that in the negative cases (30.3 +/- 15.4 vs. 11.9 +/- 9.2; p < 0.05). However, the TUNEL labeling index was not affected by the expression of those proteins. CONCLUSIONS: This study suggested that tumor progression depends on a disturbance in the cell kinetic balance caused not by a decrease in the absolute amount of cell loss but in the chaotic balance between cell loss and cell proliferation.

Preparation of phosphorus-containing silica glass microspheres for radiotherapy of cancer by ion implantation.

A chemically durable glass microsphere containing a large amount of phosphorus is useful for in situ irradiation of cancers, since they can be activated to be a beta-emitter with a half-life of 14.3 d by neutron bombardment. When the activated microspheres are injected to the tumors, they can irradiate the tumors directly with beta-rays without irradiating neighboring normal tissues. In the present study, P+ ion was implanted into silica glass microspheres of 25 microm in average diameter at 50 keV with nominal doses of 2.5 x 10(16) and 3.35 x 10(1)6 cm(-2). The glass microspheres were put into a stainless container and the container was continuously shaken during the ion implantation so that P+ ion was implanted into them uniformly. The implanted phosphorus was localized in deep regions of the glass microsphere with the maximum concentration at about 50 nm depth without distributing up to the surface even for a nominal dose of 3.35 x 10(16) cm(-2). Both samples released phosphorus and silicon into water at 95 degrees C for 7 d. On the basis of the previous study on P+-implanted silica glass plates, the silica glass microspheres containing more phosphorus which is desired for actual treatment could be obtained, without losing high chemical durability, if P+ ion would be implanted at higher energy than 50 keV to be localized in deeper region.

Heterotopic gastric mucosa in a gallbladder with an anomalous union of the pancreatobiliary duct: a case report.

A 25 year-old woman experienced a sudden onset of epigastralgia with nausea, and consulted our hospital. Because the abdominal pain did not subside with medication, she was hospitalized. On physical examination she had a slight tenderness of the right upper abdominal quadrant. Laboratory studies disclosed increases in the serum alkaline phosphatase, glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, and serum amylase levels. Abdominal ultrasonography, computed tomography, and endoscopic retrograde cholangiopancreatography revealed choledocholithiasis and a pancreatic duct which originated from the common bile duct. A common bile duct stone was removed with a basket catheter after an endoscopic sphincterotomy was performed. Since an anomalous union of a pancreatobiliary duct is a high risk factor of gallbladder cancer, laparoscopic cholecystectomy was perfomed. The post-operative course was uneventful and she was discharged on the twentieth post-operative day. In a microscopical examination of the resected specimen, a pyloric type gastric mucosa was clearly evident in the submucosa, while the remaining gallbladder demonstrated chronic cholecystitis. Some cases of heterotopic gastric mucosa in the gallbladder come from metaplasia, and metaplasia is also one of the most important factors in the carcinogenesis of gallbladder cancer. In conclusion, the present case is the first report of gastric mucosa with an anomalous union of the pancreatobiliary duct. Heterotopic gastric mucosa in the gallbladder may be one of the causes of gallbladder cancer, and close attention should, therefore, be paid to any occurrence of heterotopic gastric mucosa in this region.