Acetic acid sclerotherapy for treatment of biliary leak from an isolated right posterior sectoral duct after cholecystectomy.

Bile duct injury is one of the most serious complications of both laparoscopic and open cholecystectomy. Isolated bile duct injury can occur from the misidentification of aberrant right hepatic ducts, and it is troublesome because the early diagnosis is easy to miss and the definite treatment is controversial. We report a case of an isolated right posterior sectoral duct injury following cholecystectomy managed successfully with acetic acid sclerotherapy combined with coil embolization for a fistula tract.

Ubiquitin specific protease 19 involved in transcriptional repression of retinoic acid receptor by stabilizing CORO2A.

Deubiquitination via deubiquitinating enzymes (DUBs) has been emerged as one of the important post-translational modifications, resulting in the regulation of numerous target proteins. In this study, we screened new protein biomarkers for adipogenesis, and related studies showed that ubiquitin specific protease 19 (USP19) as a DUB is gradually decreased during adipogenesis and it regulates coronin 2A (CORO2A) as one of the components for the nuclear receptor co-repressor (NCoR) complex in some studies. The regulation of CORO2A through the deubiquitinating activity of USP19 affected the transcriptional repression activity of the retinoic acid receptor (RAR), suggesting that USP19 may be involved in the regulation of RAR-mediated adipogenesis.

Cholangiocarcinoma in choledochal cyst after cystoenterostomy: how a mistreated choledochal cyst can progress to malignancy.

This case report presents an unusual case of cholangiocarcinoma arising nearly 35 years after cystoduodenostomy for choledochal cyst. The patient visited our hospital with dyspepsia and studies revealed bezoar within the choledochal cyst caused by bile and food reflux. The patient underwent pancreaticoduodenectomy and a biopsy revealed adenocarcinoma, stage IIB. After 19 months, the patient has no recurrence to date and has recovered well. This case shows that proper surgical management and meticulous, long-term follow-up is imperative for patients with congenital choledochal cyst.

Correlation of 15-prostagladin dehydrogenase expression with clinicopathological factors and survival rate in gastric adenocarcinoma.

INTRODUCTION: The prostaglandin (PG) E2 level, which is associated with oncogenesis, progression and metastasis in various types of cancer, is determined by reciprocal regulation of 15-prostaglandin dehydrogenase (15-PGDH) and cyclooxygenase-2. This study investigated 15-PGDH expression in gastric adenocarcinoma, the associations between 15-PGDH expression and clinicopathological factors, and the correlation between 15-PGDH expression and the 5-year gastric-cancer-specific survival rate (5-year GCSS). METHODS: From 175 patients who underwent gastrectomy, we obtained biopsies of gastric adenocarcinoma tissues and adjacent normal tissues for preparation as formalin-fixed, paraffin-embedded specimens and conducted an immunohistochemical analysis. RESULTS: 15-PGDH expression was low in 65.1% of cases. 15-PGDH expression showed no relationship with age or gender, but was significantly correlated with the pathologic type, T stage, N stage, TNM stage, positive lymph node metastasis, metastasis to a larger quantity of lymph nodes, positive lymphatic invasion, positive vascular invasion, positive perineural invasion, and palliative gastrectomy. The 5-year GCSS of the low-expression group was 77.19% and a lower level of 15-PGDH expression correlated to a lower 5-year GCSS. 15-PGDH expression significantly influenced the 5-year GCSS on univariate but not multivariate analysis. CONCLUSION: Our findings indicate that 15-PGDH expression was low in gastric adenocarcinoma and was correlated with the clinicopathological factors associated with prognosis and a more advanced stage of gastric adenocarcinoma. Also, 15-PGDH expression was significantly associated with the 5-year GCSS, but was not an independent prognostic factor thereof.

Comparison of Hepatic Resection and Radiofrequency Ablation for the Treatment of Colorectal Liver Metastasis.

The liver is the major site of metastasis of primary colorectal cancer. Hepatic resection (HR) is considered the standard treatment for colorectal liver metastasis. In high-risk cases, radiofrequency ablation (RFA) can be attempted as an alternative treatment. This study compared the clinical profiles and overall and disease-free survival rates of patients with colorectal liver metastasis undergoing HR and RFA. From 1995 to 2009, we retrospectively analyzed clinical experiences of 43 and 17 patients who had undergone HR and RFA for primary colorectal cancer, respectively. To compare outcomes, we investigated the 3-year overall and disease-free survival rates. The 3-year overall survival rates of patients treated with HR and RFA were 53.5 and 47.1 %, respectively (p = 0.285); the disease-free survival rates were 35.0 and 26.9 %, respectively (p = 0.211). In the HR and RFA groups, 30 (60.2 %) and 13 (76.5 %) patients developed recurrence, respectively (p = 0.604). In the HR group, 1 patient died from postoperative liver failure, and 9 (20.9 %) developed postoperative complications, including wound infection, biliary leakage, intra-abdominal abscess, and pneumonia. In the RFA group, 1 patient (5.9 %) required prolonged inpatient care because of a procedure-related liver abscess. Although HR should be considered the first option for colorectal liver metastasis, RFA can be regarded as a primary treatment modality depending on the patient's characteristics, especially when a patient refuses surgery or has comorbidities.

Recurrence of gastric cancer in the jejunal stump after radical total gastrectomy.

This is a very rare case of the recurrence of gastric cancer in the jejunal stump after radical total gastrectomy with Roux-en-Y reconstruction. In January 2008, a 65-year-old man underwent radical total gastrectomy with Roux-en-Y reconstruction for stage IB gastric cancer of the upper body. At a follow-up in December 2011, the patient had a recurrence of gastric cancer on gastroduodenal fibroscopy. The gastroduodenal fibroscopic biopsy specimens show a well-differentiated tubular adenocarcinoma. Computed tomography showed no lymphadenopathy or hepatic metastases. At laparotomy, there was a tumor in the jejunal stump involving the pancreatic tail and spleen. Therefore, the patient underwent jejunal pouch resection, distal pancreatectomy and splenectomy. The patient was diagnosed with gastric cancer on histopathological examination.

Neuroendocrine carcinoma of the stomach: A case report.

Neuroendocrine carcinoma (NEC) is a rare tumor, comprising < 1% of stomach cancers. A 55-year-old woman was referred to our hospital with biopsy-proven gastric cancer. A shallow ulcerative lesion was detected in the lesser curvature of the lower body. It was suspected to be early gastric cancer IIA + IIC type. Thus, endoscopic submucosal dissection was performed. She was subsequently diagnosed with NEC, which is aggressive and carries a poor prognosis. We conducted a radical resection and a laparoscopic-assisted distal gastrectomy. The tumor had infiltrated the subserosal layer and 6/42 lymph nodes were involved. The mitotic index was 16/10 high power fields and the Ki-67 labeling index was 26%-50%. The final diagnosis of NEC was made according to the World Health Organization 2010 criteria. She was suspected of having jumping metastasis to the proximal margin. The patient was treated with an oral anticancer drug (5-flurouracil based drug) for 2 years. The patient has been followed up for 3 years without recurrence.

Chrysanthemum zawadskii extract protects osteoblastic cells from highly reducing sugar-induced oxidative damage.

In this study, Chrysanthemum zawadskii extract (CZE) was investigated to determine its effects on 2-deoxy-D-ribose (dRib)-induced oxidative damage and cellular dysfunction in the MC3T3-E1 mouse osteoblastic cell line. Osteoblastic cells were treated with the highly reducing sugar, dRib, in the presence or absence of CZE. Cell viability, apoptosis and reactive oxygen species (ROS) production were subsequently examined. It was observed that dRib reduced cell survival, while it markedly increased the intracellular levels of ROS and apoptosis. However, pre-treatment of the cells with CZE attenuated all the dRib-induced effects. The antioxidant, N-acetyl-L-cysteine (NAC), also prevented dRib-induced oxidative cell damage. In addition, treatment with CZE resulted in a significant increase in alkaline phosphatase (ALP) activity and collagen content, as well as in the expression of genes associated with osteoblast differentiation [ALP, collagen, osteopontin (OPN), osteoprotegerin (OPG), bone sialoprotein (BSP), osteocalcin (OC) and bone morphogenetic protein (BMP)2, BMP4 and BMP7]. In mechanistic studies of the antioxidative potential of CZE, we found that CZE reversed the dRib-induced decrease in the expression of phosphatidylinositol 3-kinase (PI3K) and protein kinase B (AKT)1 and AKT2 genes, which are master regulators of survival-related signaling pathways. CZE also upregulated the gene expression of the antioxidant enzymes, superoxide dismutase (SOD)2, SOD3 and glutathione peroxidase 4 (GPx4), which was inhibited by dRib. Taken together, these results suggest that CZE attenuates dRib-induced cell damage in osteoblastic cells and may be useful for the treatment of diabetes-associated bone disease.

Glutamate-ammonia ligase and reduction of G0 population in PANC-1 cells.

In our previous study, we screened and isolated genes that were up-regulated after partial pancreatectomy using transcriptomic analysis and glutamate-ammonia ligase (GLUL) was selected for further study based on its effect on differentiation and proliferation. In the immunohistochemical analysis, GLUL was highly up-regulated in the acinar cells and the ductal cells in the pancreas damaged through partial pancreatectomy. Overexpression of GLUL enhanced the proliferation of PANC-1 cells and INS-1 cells. GLUL overexpression shifted the major population of PANC-1 cells from the G0/G1 phase to G2/M phase. In the double thymidine blocking analysis, similar cycle duration was observed between mock cells and GLUL-overexpressing cells while GLUL-overexpressing cells were partially resistant to thymidine blocking. In the FACS analysis of cells stained with Pyronin Y and Hoechst 33342, GLUL-overexpressing cells showed lower population of cells in the G0-quiescent phase than mock cells (5-12%). In addition, GLUL-overexpressing cells had high activation levels of AKT, ERK1/2, JNK, PCNA, c-FOS, and P70S6K in PANC-1 cells. Taken together, these results suggest that GLUL contributes to pancreatic regeneration.

Long-term survival after resection of hepatocellular carcinoma.

BACKGROUNDS/AIMS: Although recent advances in surgical techniques and alternative treatment, the long-term survival >5 years after liver resection for hepatocellular carcinoma (HCC) is still unsatisfactory due to the high recurrence rate compared with other solid organ cancers. This study was conducted to analyze long-term survival after HCC resection and to develop an optimal strategy to achieve long-term survival. METHODS: A retrospective review was performed for HCC patients who underwent liver resection between 1996 and 2006. The survival rates and prognostic factors were assessed. The clinical and pathological factors of patients who survived more than 5 years were compared with those of patients whose survival was less than 5 years. The clinicopathological features characterizing long-term survivors were also reviewed. RESULTS: The overall and disease-free 5-year survival rates of 87 cases were 38.5% and 29.4%, respectively. Twenty-seven of 87 patients survived longer than 5 years after liver resection. The univariate analysis revealed that hepatitis C, the serum aspartate sminotransferase (AST) level, liver cirrhosis, Edmondson-Steiner grade, AJCC stage, and vascular invasion were significant factors for overall survival, and serum AST level, liver cirrhosis, Edmondson-Steiner grade, AJCC stage, and vascular invasion were the affecting factors for disease-free survival. In multivariate analysis, serum AST level, hepatitis C and vascular invasion were related with the overall survival, liver cirrhosis and vascular invasion which were associated with disease-free survival. Vascular invasion, AJCC stage, and the Edmondson-Steiner grade were significant factors in long-term survivors. CONCLUSIONS: Patients without liver cirrhosis, vascular invasion and normal liver function, good differentiation and an early stage may be expected to have a long-term survival.

LCP1 up-regulated by partial pancreatectomy supports cell proliferation and differentiation.

The pancreas is the critical organ controlling blood glucose levels and has been shown to rapidly regenerate after injury. In this study, 60% partial pancreatectomy (PPX) was performed on rats and the protein expression profile was acquired using 2-dimensional gel electrophoresis (2-DE)/MALDI-TOF analysis. In total, 34 proteins were shown to be up-regulated and 27 proteins were down-regulated after PPX. The up-regulated proteins were found to be involved in inflammation and the down-regulated proteins were involved in energy metabolism. Then, we compared the results from previous 4 different omics studies along with our present data and listed several genes which were found to be reproducibly regulated by PPX. The quantification of differentially regulated genes at transcriptional level by real-time PCR analysis showed that the three genes (Apoa1, Lcp1 and Lipa) were up-regulated and three genes (Gatm, Ivd and Pck2) were down-regulated. Of these, lymphocyte cytosolic protein 1 (LCP1) was highly (folds = 8.40 ± 2.57) up-regulated by PPX and found to augment cell proliferation in PANC-1 and INS-1 cells. Finally, the validation of islet markers on exogenously expressed LCP1 cells showed up-regulation of genes which are responsible for pancreatic regeneration. These data indicate that the LCP1 may play a critical role in the pancreas regeneration.

Metabolism of 1'- and 4-hydroxymidazolam by glucuronide conjugation is largely mediated by UDP-glucuronosyltransferases 1A4, 2B4, and 2B7.

Midazolam undergoes oxidative hydroxylation by CYP3A to its metabolites, which are excreted mainly as glucuronidated conjugates into the urine. In this study, we examined the glucuronidation of hydroxymidazolam in human liver microsomes (HLMs) and characterized the UDP-glucuronosyltransferases (UGTs) involved in 1'- and 4-hydroxymidazolam glucuronidation. Among the 12 UGT isoforms tested, the O- and N-glucuronidation of 1'-hydroxymidazolam was mediated by UGT2B4/2B7 and 1A4, respectively. In contrast, the glucuronidation of 4-hydroxymidazolam was mediated by UGT1A4. Consistent with these observations, the UGT1A4 inhibitor hecogenin and the UGT2B7 substrate diclofenac potently inhibited the N- and O-glucuronidation of 1'-hydroxymidazolam in HLMs, respectively. A correlation analysis of UGT enzymatic activity and the formation rate of glucuronide metabolites from 1'- and 4-hydroxymidazolam in 25 HLMs showed that hydroxymidazolam glucuronidation is correlated with UGT1A4-mediated lamotrigine glucuronidation and UGT2B7-mediated diclofenac glucuronidation activity. Taken together, these findings indicate that UGT1A4, 2B4, and 2B7 are major isoforms responsible for glucuronide conjugate formation from 1'- and 4-hydroxymidazolam, which are the two major oxidative metabolites of midazolam.

Isolation of genes involved in pancreas regeneration by subtractive hybridization.

The deterioration of ß cells in the pancreas is a crucial factor in the progression of diabetes mellitus; therefore, the recovery of ß cells is of vital importance for effective diabetic therapeutic strategies. Partially pancreatectomized rats have been used for the investigation of pancreatic regeneration. Because it was determined that tissue extract from the partially-dissected pancreas induces pancreatic differentiation in embryonic stem cells, paracrine factors were thought to be involved in the regeneration. In this study, we screened for genes that had higher mRNA levels 2 days after 60%-pancreatectomy. The genes were isolated using subtractive hybridization and DNA sequencing. Twelve genes (adipsin, Aplp2, Clu, Col1a2, Glul, Krt8, Lgmn, LOC299907, LOC502894, Pla2g1b, Reg3α and Xbp1) were identified, and RT-PCR and real-time PCR analyses were performed to validate their expression levels. Among the genes identified, three genes (Glul, Lgmn and Reg3a) were selected for further analyses. Assays revealed that Glul and Reg3α enhance cell growth. Glul, Lgmn and Reg3α change the expression level of islet marker genes, where NEUROD, NKX2.2, PAX4 and PAX6 are up-regulated and somatostatin is down-regulated. Thus, we believe that Glul, Lgmn and Reg3a can serve as novel targets in diabetes mellitus genetic therapy.

[Expressions and clinical significances of c-met, c-erbB-2, COX-2, and IL-6 in the biliary tract cancers].

BACKGROUND/AIMS: c-met, c-erbB-2, interleukin (IL)-6, and cyclooxygenase (COX)-2 expressions are considered to be implicated in the carcinogenesis and progression of cholangiocarcinoma, but the molecular pathogenesis of cholangiocarcinoma is still poorly understood. We aimed to analyze the expressions of each marker and their relationships with clinicopathologic factors. METHODS: One hundred and fourteen tissue samples were obtained from surgically resected specimens from patients with biliary tract cancer. The expressions of c-met, c-erbB-2, COX-2, and IL-6 were examined by immunohistochemically. The expression of each marker and correlations between these markers and clinicopathologic factors were analyzed. RESULTS: The expression rates of each maker were as follows: c-met 34/112 (30.4%), c-erbB-2 5/112 (4.5%), COX-2 53/113 (46.9%), and IL-6 68/113 (60.2%), respectively. c-met expression was more frequently observed in cases with invasion through the adjacent connective tissues (p=0.0263). IL-6 overexpression was more frequently observed in cases with absent lymph node metastasis (p=0.0325). Either c-erbB-2 expression or COX-2 expression was significantly associated with lymph node metastasis (p=0.0442). CONCLUSIONS: The expression of c-met was closely related to the invasiveness of cholangiocarcinoma. Co-expression of c-met, COX-2 and, IL-6 showed a significant correlation with invasiveness and lymph node metastasis and these could be useful marker to guide clinical outcome in patients with cholangiocarcinoma.

Characterization of ebastine, hydroxyebastine, and carebastine metabolism by human liver microsomes and expressed cytochrome P450 enzymes: major roles for CYP2J2 and CYP3A.

Ebastine undergoes extensive metabolism to form desalkylebastine and hydroxyebastine. Hydroxyebastine is subsequently metabolized to carebastine. Although CYP3A4 and CYP2J2 have been implicated in ebastine N-dealkylation and hydroxylation, the enzyme catalyzing the subsequent metabolic steps (conversion of hydroxyebastine to desalkylebastine and carebastine) have not been identified. Therefore, we used human liver microsomes (HLMs) and expressed cytochromes P450 (P450s) to characterize the metabolism of ebastine and that of its metabolites, hydroxyebastine and carebastine. In HLMs, ebastine was metabolized to desalkyl-, hydroxy-, and carebastine; hydroxyebastine to desalkyl- and carebastine; and carebastine to desalkylebastine. Of the 11 cDNA-expressed P450s, CYP3A4 was the main enzyme catalyzing the N-dealkylation of ebastine, hydroxyebastine, and carebastine to desalkylebastine [intrinsic clearance (CL(int)) = 0.44, 1.05, and 0.16 microl/min/pmol P450, respectively]. Ebastine and hydroxyebastine were also dealkylated to desalkylebastine to some extent by CYP3A5. Ebastine hydroxylation to hydroxyebastine is mainly mediated by CYP2J2 (0.45 microl/min/pmol P450; 22.5- and 7.5-fold higher than that for CYP3A4 and CYP3A5, respectively), whereas CYP2J2 and CYP3A4 contributed to the formation of carebastine from hydroxyebastine. These findings were supported by chemical inhibition and kinetic analysis studies in human liver microsomes. The CL(int) of hydroxyebastine was much higher than that of ebastine and carebastine, and carebastine was metabolically more stable than ebastine and hydroxyebastine. In conclusion, our data for the first time, to our knowledge, suggest that both CYP2J2 and CYP3A play important roles in ebastine sequential metabolism: dealkylation of ebastine and its metabolites is mainly catalyzed by CYP3A4, whereas the hydroxylation reactions are preferentially catalyzed by CYP2J2. The present data will be very useful to understand the pharmacokinetics and drug interaction of ebastine in vivo.