[A case of aortic thrombus during chemotherapy for esophageal cancer].

A 59-year-old man presented to our hospital with a chief complaint of epigastric pain. Pertinent history included a distal gastrectomy for gastric cancer and alcohol dependence. He underwent contrast-enhanced computed tomography (CT) and esophagogastroduodenoscopy, which led to a diagnosis of esophageal cancer (cT2N2M1, stage IVb). Subsequently, he underwent chemotherapy using 5-fluorouracil and cis-diamminedichloroplatinum and radiotherapy. A total of 44 days after treatment initiation, the patient experienced nausea and hepatobiliary enzyme elevation. CT and abdominal ultrasonography were performed, and he was diagnosed with an abdominal aortic thrombus. Intravenous heparin was administered as an anticoagulant therapy. Twenty-two days after treatment initiation, the thrombus was no longer visible on abdominal ultrasonography. The patient was then treated with warfarin. It cannot be ruled out that the patient's hepatobiliary enzyme elevation was induced by the anticancer drugs. However, enzyme elevation improved with the disappearance of the abdominal aortic thrombus, suggesting that the aortic thrombus may have contributed to the hepatobiliary enzyme elevation. No thrombus recurrence was observed until the patient's death after an initial treatment with antithrombotic agents. This case indicates that malignant tumors and chemotherapy can cause aortic thrombi, and thus, care should be exercised in monitoring this potential complication.

[A case of ischemic proctitis caused by impaired venous blood flow 11 months after surgery for sigmoid colon cancer].

A Japanese male in his 50s was presented to our hospital with the chief complaint of positive fecal immunochemical test. He had a history of hypertension. He underwent colonoscopy and was diagnosed with sigmoid colon cancer. He also underwent laparoscopic sigmoid colectomy with D3 lymph node dissection for sigmoid colon cancer. The inferior mesenteric artery and inferior mesenteric vein were amputated at the root of the vessels. The patient received adjuvant chemotherapy and was recurrence-free. Eleven months after the surgery, lower abdominal pain during defecation appeared. Contrast-enhanced computed tomography (CT) and colonoscopy showed marked rectal mucosal edema and increased fatty tissue density (dirty fat sign) around the anorectal side of the anastomosis. Intestinal blood flow was maintained. There were many fine blood vessels around the rectal wall, and the amputated distal part of the superior rectal artery was retrogradely contrasted. Amputated superior rectal artery and superior rectal vein were dilated than before. Colonoscopy revealed mucosal redness, edema, and easy bleeding on the anorectal side of the anastomosis. Abdominal contrast-enhanced 3D-CT showed increased arterial blood flow and increased fine blood vessels around the rectal wall. It suggested the presence of an arteriovenous fistula and venous congestion. Conservative treatment with total parenteral nutrition and prednisolone infusion did not improve the patient's condition, and a colostomy was performed. After colostomy, the pain improved, and the CT scan of the abdomen showed improvement in arterial blood flow and venous congestion. Colostomy was closed after 10 months. There has been no relapse since the closure of the colostomy. There are few reports on ischemic proctitis on the anorectal side of the anastomosis after colon cancer resection due to impaired venous blood flow.

[A case of young hepatocellular carcinoma with non-B, non-C, non-cirrhotic background].

A man in his 20s visited a local physician because of upper abdominal pain, and an abdominal ultrasonography revealed hepatic tumors. He was then referred to our hospital. The patient had no history of blood transfusion, tattoos, habitual alcohol consumption, or narcotic drug use. Physical examination revealed abdominal fullness. Biochemical tests were negative for hepatitis virus markers and autoantibodies. Liver enzyme levels were high;further, the levels of alpha-fetoprotein and protein induced by vitamin K absence or antagonist-II (PIVKA-II) were elevated. Chest and abdominal dynamic enhanced computed tomography and magnetic resonance imaging scans showed multiple lung tumors and multiple liver tumors. An arterial phase contrast-enhanced computer tomography image showed multiple nodular heterogeneous hyperattenuating masses with washout in the equilibrium phase. A huge mass in the right hepatic lobe had a large area of central necrosis. We suspected hepatocellular carcinoma or undifferentiated mesenchymal tumor. Liver biopsy showed moderately differentiated hepatocellular carcinoma without fibrosis in the background liver. This patient was diagnosed with hepatocellular carcinoma that developed in a normal liver. The patient was treated with molecular-targeted drugs. Tumor enhancement decreased;however, the tumor size remained unchanged. The patient lived for 9 months. A search using the retrieval terms "non-hepatitis B virus/non-hepatitis C virus", "non-cirrhotic", "young adult", and "hepatocellular carcinoma" revealed 12 case reports in the Igaku Chuo Zasshi database. Many cases had multiple tumors that were large in size as well as had venous invasion, and surgeries were performed because liver functions were normal. The present case is noteworthy because hepatocellular carcinoma with a non-hepatitis B virus/non-hepatitis C virus and non-cirrhotic background in a young patient is rare.

[A case of corrosive gastritis caused by accidental ingestion of calcium chloride that resulted in gastric stenosis and penetration].

A Japanese woman in her 40s came to our emergency room with vomiting and upper abdominal pain after drinking a bottle of milk tea at home. She had a history of bipolar disorder. Blood tests revealed hypercalcemia (calcium level of 18.6mg/dl). Abdominal computed tomography depicted thickening of the gastric wall and hyperabsorbed material in the stomach. Upper gastroduodenal endoscopy showed extreme mucosal redness from the gastric body to the pylorus. The hypercalcemia improved with intravenous infusion of zoledronic acid. The patient had not been taking any medication that could have caused hypercalcemia. Later, her father drank the same bottle of milk tea at home and developed upper abdominal pain. He was admitted to the hospital because of vomiting, and computed tomography showed hyperabsorbed material in the stomach, as in his daughter's case. Computed tomography of the bottle of milk tea revealed a highly absorbent substance. The bottle was sent to the forensics laboratory for testing, and it was found to contain calcium chloride. Thus both patients had consumed a beverage containing calcium chloride, and corrosive gastritis was diagnosed. Despite fasting and intravenous drip therapy, the first patient underwent a total gastrectomy because of severe stenosis and perforation of the gastric lumen.

[A case of leptomeningeal carcinomatosis involving loss of eyesight after total gastrectomy for gastric cancer].

A 78-year-old woman had undergone total gastrectomy and chemotherapy for gastric cancer (pT4N3bM0 Stage IIIC, poorly differentiated adenocarcinoma). She received S-1 monotherapy 3 times weekly (S-1 at 80mg twice daily for 14 days, every 3 weeks). She underwent routine examinations, including tumor markers and computed tomography. She had no signs of recurrent disease, but she suffered from a loss of eyesight 2 years and 8 months after the operation. A choked disc was found, but she had no headaches, nausea, or unconsciousness, which indicated high intraventricular pressure. Enhanced T2-weighted magnetic resonance imaging showed high intensity around the optic nerve. We performed cerebrospinal fluid cytological analysis, which showed poorly differentiated adenocarcinoma. She was diagnosed as having leptomeningeal carcinomatosis of gastric cancer. The patient chose best supportive care and died 2 months after symptoms appearance. Histological analysis during the autopsy showed moderately to poorly differentiated adenocarcinoma. The carcinoma had also infiltrated the spinal cord, peritoneum, and adrenal glands. Histologically, the carcinoma had infiltrated the optic nerve, which caused loss of eyesight. We have not yet established effective therapies for leptomeningeal carcinomatosis, and the prognosis is poor. Leptomeningeal carcinomatosis of gastric cancer that appears by loss of eyesight is very rare. This case illustrates that the possibility of leptomeningeal carcinomatosis should be considered when we treat patients with loss of eyesight of an unknown cause after surgery.

A case report of adenosquamous cell carcinoma of the duodenal papilla.

Adenosquamous carcinoma of the duodenal papilla is rare. A 73-year-old man was referred to the Saiseikai-Matsusaka General Hospital with upper abdominal pain and liver dysfunction. Computed tomography (CT) revealed dilatation of the common bile duct (CBD) and intrahepatic bile duct along with a tumor in the distal CBD. The tumor showed enhancement in the arterial phase on contrast-enhanced CT. We performed endoscopic retrograde cholangiopancreatography and noted a red, erosive, bleeding mass in the duodenal papilla with obstruction of the distal CBD, and dilatation of the CBD. Histopathological inspection of a biopsy of the duodenal papilla showed a mixture of adenocarcinoma and squamous cell carcinoma, suggesting the presence of adenosquamous cell carcinoma in the duodenal papilla. Abdominal examinations including positron emission tomography/CT showed no metastasis or lymph node swelling. The clinical stage was determined to be cT2N0M0 Stage IB. We performed subtotal stomach-preserving pancreaticoduodenectomy. Histopathological inspection of the specimen showed a mixture of adenocarcinoma and squamous cell carcinoma, and squamous cell carcinoma accounted for 40% of the tumor. The tumor was defined as pathological Stage IIA, AcbBd, mixed type, med, pT3b, sci, INFb, ly2, v1, ne2, pN1, HM0, PM0, EM0, PV0, A0, R0, pT3N0M0. We suggested adjuvant chemotherapy, but the patient declined adjuvant chemotherapy and wished to be discharged. Abdominal ultrasonography revealed multiple liver metastases 3 months postoperatively. The patient opted for best supportive care and died 9 months postoperatively. Examination of 23 reports of adenosquamous cell carcinoma of the duodenal papilla in Japan suggested that adenosquamous cell carcinoma of the duodenal papilla has a poorer prognosis compared with adenocarcinoma of the duodenal papilla. Some reports have stated that the growth rate is faster for squamous cell carcinoma than for adenocarcinoma. In our case, the tumor was enhanced in the arterial phase and this represents a feature of adenosquamous cell carcinoma of the duodenal papilla. Chemotherapy has not been established for adenosquamous cell carcinoma of the duodenal papilla. We are confident that we can establish effective chemotherapies in the future.

A case of ulcerative colitis with a variety of autoimmune diseases including ankylosing spondylitis, type 2 autoimmune pancreatitis, and primary sclerosing cholangitis.

Ankylosing spondylitis (AS), primary sclerosing cholangitis (PSC), and autoimmune pancreatitis (AIP) are known as extraintestinal manifestations (EIMs) of ulcerative colitis (UC). A 74-year-old Japanese man visited our hospital because of white stool. He had been diagnosed with AS when he was 30 years old, and he was HLA-B27-positive. Based on various examination results, it was suspected that AIP had caused bile duct stricture. During the clinical course, he was diagnosed with UC and PSC. Then, AIP was diagnosed because he had localized pancreatic enlargement, irregular stenosis of the main pancreatic duct, PSC, and no tumor cells of pancreas. A patient with all four of these diseases, AS, AIP, PSC, and UC, is very rare. Therefore, we report a quite rare case with three EIMs (AS, PSC, and AIP) of UC.

Role of poly (A) tail as an identity element for mRNA nuclear export.

Different RNA species are rigorously discriminated and exported by distinct export factors, but this discrimination mechanism remains largely unknown. We previously showed, by RNA microinjection experiments, that intronless mRNAs are discriminated from U snRNAs based on their difference in RNA length. However, it was unclear how they are discriminated in the natural situation in which their nascent transcripts emerge progressively during transcription. We hypothesized that transcription from the corresponding promoters is important for this discrimination. Here we show that contrary to our hypothesis, the discrimination process was not significantly influenced by whether transcription occurred from an mRNA- versus a U snRNA-type promoter. Rather, the features of transcribed RNAs determined the RNA identity, consistent with our previous results of RNA microinjection. Moreover, we found that the poly (A) tail can function as an identity element for mRNA export. The presence of a poly (A) tail of an appropriate length committed otherwise short Pol II transcripts to the mRNA export pathway in a dominant manner, indicating that the poly (A) tail either contributes to increasing the RNA length or functions as a platform to recruit mRNA export factors. Our results reveal a novel function of the poly (A) tail in mRNA export.

Radiofrequency ablation combined with chemoembolization for the treatment of hepatocellular carcinomas larger than 5 cm.

PURPOSE: To evaluate survival, recurrence-free survival, technical success, technique effectiveness, and safety of radiofrequency (RF) ablation combined with chemoembolization in patients with hepatocellular carcinomas (HCCs) larger than 5 cm. MATERIALS AND METHODS: Patients with Child-Pugh class A or B cirrhosis and three or fewer HCCs with a maximum tumor diameter of 5.1-10 cm were included. Twenty patients with 32 HCCs were included. There were 16 men and four women with mean age of 69 years +/- 7.4 (range, 46-79 years).The maximum mean tumor diameter was 6.2 cm (range, 5.1-9.5 cm). RF ablation was performed under computed tomographic (CT) fluoroscopic guidance 1-2 weeks after chemoembolization. The primary endpoint of this study was survival. RESULTS: RF electrodes were placed in the planned sites, and RF ablation was completed with a planned protocol (technical success rate, 100%). Tumor enhancement was eradicated in all patients after 32 RF sessions. The primary and secondary technique effectiveness rates were 40% and 100%, respectively. There were two major complications in the 32 RF sessions (6%)--hepatic abscess and diaphragm perforation. Local tumor progression developed in five of the 20 patients (25%) during the mean follow-up of 30 months. The overall and recurrence-free survival rates were, respectively, 100% and 74% at 1 year, 62% and 28% at 3 years, and 41% and 14% at 5 years. The serum bilirubin level of 1.0 mg/dL (17.1 micromol/L) or less was a significantly better prognostic factor in the univariate analysis. CONCLUSIONS: This combination therapy may enhance survival in patients with HCCs larger than 5 cm.

Early-stage hepatocellular carcinoma: radiofrequency ablation combined with chemoembolization versus hepatectomy.

PURPOSE: To retrospectively evaluate the long-term results of radiofrequency (RF) ablation combined with chemoembolization (combination therapy) as compared with hepatectomy for the treatment of early-stage hepatocellular carcinoma (HCC). MATERIALS AND METHODS: The study was approved by the institutional review board, and informed consent was waived. Patients with early-stage HCC were included if they underwent either combination therapy or hepatectomy and met the following inclusion criteria: no previous treatment for HCC, three or fewer tumors with a maximum diameter of 3 cm or less each or a single tumor with a maximum diameter of 5 cm or less, Child-Pugh class A liver profile, no vascular invasion, and no extrahepatic metastases. The primary endpoint was overall survival, and the secondary endpoint was recurrence-free survival. RESULTS: One hundred four patients (mean age, 66.5 years +/- 8.7 [standard deviation]; 79 men, 25 women) underwent combination therapy, and 62 patients (mean age, 64.5 years +/- 9.6; 51 men, 11 women) underwent hepatectomy. The 1-, 3-, and 5-year overall survival rates following combination therapy (98%, 94%, and 75%, respectively) were similar (P = .87) to those following hepatectomy (97%, 93%, and 81%, respectively). The 1-, 3-, and 5-year recurrence-free survival rates were also comparable (P = .70) for combination therapy (92%, 64%, and 27%, respectively) and hepatectomy (89%, 69%, and 26%, respectively). CONCLUSION: RF ablation combined with chemoembolization in patients with early-stage HCC provides overall and disease-free survival rates similar to those achieved by hepatectomy.

Dual topology of functional Toll-like receptor 3 expression in human hepatocellular carcinoma: differential signaling mechanisms of TLR3-induced NF-kappaB activation and apoptosis.

Toll-like receptor 3 (TLR3) is a pattern-recognizing receptor that is involved in immune signaling and plays a crucial role in survival by being able to recognize various viral components including double-stranded RNA (dsRNA). TLR3 expression and function in cancer cells are not well understood. We investigated the expression of TLR3 in hepatocellular carcinoma (HCC) cells and the function of TLR3 signaling by stimulation and transfection with polyinosinic-polycytidylic acid (Poly I:C), a synthetic form of dsRNA. TLR3 mRNA was expressed in HCC tissues as well as in non-tumor tissues. Positive immunohistochemical staining for TLR3 was observed in 52.7% of HCC tissues, and in HCC cells we found both membranous and cytoplasmic expression of TLR3. While cell surface stimulation of TLR3 with Poly I:C did not affect cell viability, it did activate NF-kappaB levels. In contrast, cytoplasmic stimulation with transfected Poly I:C significantly induced apoptosis accompanied by the down-regulation of anti-apoptotic protein. Transfected Poly I:C also synergistically augmented TRAIL-induced apoptosis, but only with low levels of transfected Poly I:C was IFN-beta production not observed. In conclusion, our results indicate that TLR3 expression in HCC plays an important role with regard to cell survival and proapoptotic activity. Endogenously expressed TLR3 may provide new clinical prospects for TLR3 agonists as cytotoxic agents in HCC.

Protective role of interleukin-18 against Fas-mediated liver injury.

Interleukin (IL)-18 plays an important role in the pathogenesis of several liver diseases as well as Fas-mediated apoptosis. However, the effects of IL-18 on Fas-mediated liver injury have not been well elucidated. Therefore, we examined the effects of IL-18 on Fas-mediated apoptosis in in vitro and in vivo experiments. We found that recombinant IL-18 protected mouse hepatocellular carcinoma cell lines, BNL5, from Fas-mediated apoptosis in a dose-dependent manner with up-regulation of both nuclear factor (NF) kappaB and X-linked inhibitors of apoptosis (XIAP). IL-18 transgenic (Tg) mice were also protected from Fas-mediated liver injury and this was further confirmed by histological study and TUNEL staining. In IL-18 Tg mice, up-regulation of XIAP and down-regulation of caspase 3 were observed after injection of anti-Fas, which was consistent with the in vitro findings. These results suggest that IL-18 suppresses Fas-mediated apoptosis of hepatocytes by up-regulation of NFkappaB and XIAP, following inhibition of caspase-3 activity. This observation raises the possibility that IL-18 could be a therapeutic strategy for Fas-mediated liver injury as a negative regulator of XIAP.

Adenovirus-mediated transfection of caspase-8 sensitizes hepatocellular carcinoma to TRAIL- and chemotherapeutic agent-induced cell death.

Caspase-8 belongs to the cysteine protease family and is known to be activated at the initial step in the cascade of TRAIL-induced apoptosis. The activation of procaspase-8 can be blocked by a relatively large amount of c-FLIP, which renders resistance to death receptor-mediated apoptosis in many types of cancer cells. To ask if extrinsic over-expression of caspase-8 contributes to the induction of apoptosis, we introduced the caspase-8 gene into HCC cells using an adenoviral (Adv) vector (Adv-Casp8). We demonstrated that Adv-Casp8 increased expression of active forms of caspase-8 in MOI-dependent manner. A large amount of Adv-Casp8 (MOI of 50) induced apoptosis significantly in HCC cells and resulted in downregulation of c-FLIP (in SK-Hep1, HLE, and HepG2 cells), XIAP, survivin, and Bcl-xL (in HLE cells) and dynamic release of cytochrome c and Smac from the mitochondria into the cytosol. On the other hand, a small amount of Adv-Casp8 (MOI of 10) causes a slight but detectable increase in the level of apoptosis with only a small effect on anti-apoptotic proteins and mitochondrial activation. However, small amounts of Adv-Casp8 augmented TRAIL- or chemotherapeutic agent-induced cell death (with an MOI of 10 or 20, respectively). These results suggest both that exogenous over-expression of caspase-8 by Adv-Casp8 may be essential for induction of HCC cell death and that the combination of Adv-Casp8 and TRAIL or chemotherapeutic agents could provide a useful strategy for treatment of HCC.

Tubulointerstitial nephritis associated with IgG4-related autoimmune disease.

Autoimmune pancreatitis is a chronic fibroinflammatory condition primarily affecting the pancreas. Recent accumulating evidence suggested that autoimmune pancreatitis is a systemic autoimmune disease (immunoglobulin G4 [IgG4]-related autoimmune disease) affecting various organs with dense infiltration of IgG4-positive mononuclear cells. Tubulointerstitial nephritis is still a mysterious disease with an unknown cause. We report 2 cases of tubulointerstitial nephritis associated with autoimmune pancreatitis. In these patients, dense infiltrations of IgG4-positive mononuclear cells were observed in renal interstitium, with high serum IgG4 levels. Furthermore, in patient 1, who had sclerosing cholangitis, serum alkaline phosphatase and serum creatinine levels changed synchronously. Steroid therapy was followed by improved renal function and serum IgG4 levels in both patients. Because tubulointerstitial nephritis associated with IgG4-related autoimmune disease shows a favorable response to steroids and the renal dysfunction and pancreatic dysfunction are reversible, awareness of this entity is necessary for early diagnosis and prompt treatment. In addition, these cases support the hypothesis that IgG4-related autoimmune disease could be one cause of tubulointerstitial nephritis.

Lamivudine for treatment of spontaneous exacerbation and reactivation after immunosuppressive therapy in patients with hepatitis B virus infection.

BACKGROUND/AIMS: We evaluated the efficacy of lamivudine therapy for treatment of spontaneous exacerbation and reactivation after immunosuppressive therapy in patients with hepatitis B virus infection. Lamivudine is an effective therapy if used in early stages of both spontaneous exacerbation and reactivation after immunosuppressive therapy. METHODOLOGY: In our study, twelve patients experienced flares of chronic hepatitis B over a three-year period. RESULTS: Three patients whose pretreatment total bilirubin levels were more than 7mg/dL died of fatal liver failure despite lamivudine therapy. CONCLUSIONS: We concluded that if pretreatment serum bilirubin levels are higher than 7 mg/dL, lamivudine therapy alone may be insufficient and more effective therapies should be considered concomitantly with lamivudine.

Corona enhancement in ultrasonographical post-vascular phase images with microbubble contrast agent: a novel specific sign for hepatocellular carcinomas.

In 1.5 Harmonic Imaging ultrasonography (1.5 HI US), images are obtained in a band intermediate between the fundamental and 2nd harmonic components, resulting in stronger contrast enhancement than in conventional harmonic imaging. We attempted to assess the hemodynamics of hepatocellular carcinomas (HCC) with special attention to blood drainage using 1.5 HI US. Forty-two HCC nodules, metastatic liver tumors and hepatic hemangiomas were studied. In contrast studies, intermittent ultrasound transmission was performed for a period of up to 45 sec after the injection of contrast agent, which was regarded as the vascular phase. The time point of 5 min later was specified as the post-vascular phase, and images were obtained by single manual transmission for comparison of contrast enhancement with surrounding hepatic parenchyma. In addition, histological examination was performed. 1.5 HI US clearly demonstrated the strong tumor vessels in most HCCs. Corona enhancement, in which the areas surrounding the tumor are enhanced, was observed in 71.4% (30/42) of HCC nodules during the post-vascular phase. This sign was not observed in any other tumors. Histological findings revealed that CD34-positive-endothelial cells were prominent in the surrounding area of HCC. In conclusion, 1.5 HI US is an effective tool for evaluating hemodynamics in both early- and post-vascular phase. Corona enhancement may be due to the trapping of contrast agent in the endothelial cells in the surround of HCC nodules and be a novel specific sign for HCC.

The cyclin-dependent kinase inhibitor flavopiridol sensitizes human hepatocellular carcinoma cells to TRAIL-induced apoptosis.

Flavopiridol was one of the first cyclin-dependent kinase inhibitors demonstrated to have an antitumor effect in several cancer types. Here, we investigated the effects of flavopiridol on TNF-related apoptosis-inducing ligand (TRAIL) in the human hepatocellular carcinoma (HCC) cell lines HLE and HepG2, and evaluated the role of flavopiridol in apoptosis. To better understand the mechanism of increased TRAIL sensitivity in HCC cells, we determined the effect of flavopiridol on cell surface expression of TRAIL and TRAIL receptors using flow cytometry analysis. The levels of survivin, FLIP, Bcl-xL and X-chromosome-linked IAP (XIAP) in treated and untreated cells was also determined. Flavopiridol decreased cell viability in a dose-dependent manner in the two HCC cell lines tested. The pan-caspase inhibitor z-VAD-FMK did not inhibit the effect. However, subtoxic levels of flavopiridol dramatically enhanced TRAIL-induced apoptosis in both cells. Flavopiridol up-regulated TRAIL, TRAIL-R1 and TRAIL-R2 in both cell lines. In addition, flavopiridol down-regulated expression of survivin in both cell lines, and expression of FLIP and Bcl-xL were down-regulated in HLE cells. In summary, flavopiridol augmented TRAIL sensitivity by up-regulation of TRAIL receptors and down-regulation of survivin, FLIP and Bcl-xL. Thus, combining flavopiridol with a TRAIL agonist may prove to be an effective new strategy for treatment of HCC.

COX-2 inhibitors sensitize human hepatocellular carcinoma cells to TRAIL-induced apoptosis.

Cyclooxygenase (COX)-2 is upregulated in a variety of human cancers, including in hepatocellular carcinoma (HCC), whereas it is undetectable in most normal tissue. Evidence suggests that COX-2 is likely to be involved in hepatocarcinogenesis and, thus, COX-2 may be involved in an early process in carcinogenesis, dedifferentiation. To address this possibility, we investigated the effect of COX-2 inhibitors on TNF-related apoptosis, inducing ligand (TRAIL) sensitivity and its molecular mechanisms, with special attention to anti-apoptotic proteins. We used the highly selective COX-2 inhibitors, NS398 and CAY10404. We also used the MTT assay and cytological analysis of DAPI-stained DNA to assess viability and apoptosis in two HCC cells (SK-Hep1 and HLE). In order to ask what led to increased sensitivity to TRAIL in HCC cells, cell surface expression of TRAIL and TRAIL-receptors was investigated using flow cytometry analysis. Expression of survivin, X-chromosome-linked IAP (XIAP), Bcl-xL, AKT and phospho-AKT was also investigated using immunoblotting. COX-2 inhibitors resulted in a concentration-dependent decrease in cell viability in the two HCC cell lines tested. Subtoxic levels of COX-2 inhibitors did not significantly augment TNFalpha-induced apoptosis but did dramatically enhance TRAIL-induced apoptosis in both cell lines. TRAIL receptor 2/death receptor 5 (TRAIL-R2/DR5) expression was significantly up-regulated in SH-Hep1 and HLE cells. TRAIL receptor 1/death receptor 4 (TRAIL-R1/DR4) expression was up-regulated only in SK-Hep1. Expression of survivin and Bcl-xL was down-regulated in SK-Hep1 and HLE cells in the presence of CAY10404 but XIAP was not affected. Expression of survivin, Bcl-xL and XIAP was down-regulated in SK-Hep1 cells in the presence of NS398. Survivin expression was also down-regulated in the presence of NS398 in HLE cells. Finally, NS398 also decreased phospho-AKT in SK-Hep1 cells. These results demonstrate that COX-2 inhibitors can induce apoptosis and augment TRAIL sensitivity by up-regulation of TRAIL receptors and down-regulation of both survivin and AKT signaling.

The usefulness of 1.5 harmonic imaging ultrasonography with Levovist in the diagnosis of focal hepatic tumors.

In contrast-enhanced 1.5 harmonic imaging sonography, images are obtained in a band intermediate between the fundamental and the 2nd harmonic components. In the present study, we investigated the usefulness of 1.5 harmonic imaging sonography with the use of the contrast agent Levovist for the diagnosis of hepatocellular carcinoma (HCC), metastatic hepatic tumor, and hepatic hemangioma. The subjects in this study were 64 patients with 70 nodules of hepatic tumors (42 nodules in 36 cases of hepatocellular carcinoma, 20 nodules in 20 cases of metastatic hepatic tumor, and 8 nodules in 8 cases of hepatic hemangioma). Contrast enhancement of tumors acquired in the early, portal, and late phases with 1.5 harmonic imaging sonography were compared to classify the tumors. 1.5 harmonic imaging sonography of HCC showed contrast enhancement of 36 nodules (85.7%). Hypervascular enhancement in the early phase, which was maintained in the portal phase, changed to images with no contrast enhancement with partial persistence of contrast enhancement in the late phase. 1.5 harmonic imaging sonography of metastatic hepatic tumor showed hypervascular enhancement of the margin of 20 nodules (100%) in the early and portal phases, which changed to images with no contrast enhancement in the late phase. 1.5 harmonic imaging sonography of hepatic hemangiomas maintained hypervascular enhancement on the tumor margin of 5 nodules (62.5%) in the early and portal phases. When early phase 1.5 harmonic imaging sonography did not show hypervascular enhancement in 3 nodules (37.5%), and late-phase images confirmed that these 3 nodules were hypervascular enhancement on the tumor margin. 1.5 harmonic imaging sonography of hepatic tumors (hepatocellular carcinoma, metastatic hepatic tumor and hepatic hemangioma) provided characteristic findings of contrast enhancement in the early, portal, and late phases, and will contribute to differential diagnosis.

Involvement of tumor necrosis factor-related apoptosis-inducing ligand and tumor necrosis factor-related apoptosis-inducing ligand receptors in viral hepatic diseases.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces apoptosis in tumor cells, but not in most normal cells. The role of TRAIL in hepatic cell death and hepatic diseases is not well understood. The present study investigated the expression of TRAIL and TRAIL receptors (TRAIL-Rs) in patients with hepatitis C virus infection using immunohistochemistry and examined physiological roles under viral infection in the HepG2 cell line. Staining of TRAIL or TRAIL-Rs was prominent in the cytoplasm and membrane of hepatocytes in the periportal area. Some liver-infiltrating lymphocytes also displayed positive staining for TRAIL. Staining intensity was significantly increased with disease progression, particularly in the periportal area. AdCMVLacZ (Q-BIOgene, Carisbad, Calif) infection was also found to induce apoptosis in HepG2 cells and significantly augment TRAIL-induced apoptosis. Anti-TRAIL antibody significantly inhibited apoptosis induced by AdCMVLacZ infection. Flow cytometry analysis revealed that both TRAIL-R2 and TRAIL were up-regulated on the cell surface of HepG2 cells with AdCMVLacZ infection. Transforming growth factor-beta1 also enhanced TRAIL expression in HepG2 cells. These results indicate that TRAIL/TRAIL-R apoptotic pathways play important roles in the hepatic cell death during viral infection.