The safety and efficacy of Ringer's solution loading with rectal diclofenac for prevention of post-endoscopic retrograde cholangiopancreatography pancreatitis: The RESOLUTION-PEP study.

Objectives: We evaluated the safety and efficacy of aggressive hydration with rectal non-steroidal anti-inflammatory drugs for the prevention of post-endoscopic retrograde cholangiopancreatography (ERCP) pancreatitis (PEP). Methods: This prospective, single-arm, multicenter trial was conducted at 12 institutions between October 2020 and August 2021. We enrolled 231 patients who had intact papillae and were scheduled to undergo ERCP. All patients were administered rectal diclofenac before ERCP. They received aggressive hydration with intravenous lactated Ringer's solution in an initial bolus of 5 ml/kg at the start of ERCP, followed by 3 ml/kg/h for 8 h after the procedure. The primary outcome was the occurrence of PEP. Secondary outcomes included PEP severity, hyperamylasemia, and adverse events. Results: The mean age of the patients was 68.8 ± 13.7 years, and 81 patients (35.1%) were 75 years or older. Thirteen patients developed PEP (5.6%, 95% confidence interval 3.0%-9.4%). There were 11 cases (4.8%) of mild pancreatitis and two cases (0.9%) of severe pancreatitis. Forty-five patients (19.5%) developed hyperamylasemia and one patient developed non-severe peripheral edema. Conclusions: Aggressive hydration combined with rectal diclofenac may be a promising strategy for the prevention of PEP. Furthermore, it is safe even for older individuals.

Second and third-look endoscopy for the prevention of post-ESD bleeding.

The efficacy of 2nd-look esophagogastroduodenoscopy (EGD) with endoscopic hemostatic therapy (EHT) for the prevention of postendoscopic submucosal dissection (ESD) clinical bleeding remains controversial. The aim of this study was to estimate post-ESD bleeding rate using 2nd and 3rd-look strategy, and to determine risk factors for clinical bleeding, and for EHT at 2nd and 3rd-look EGDs.Three hundred forty-four consecutive patients with early gastric cancer or adenoma underwent ESD from January 2006 through March 2012. Second and 3rd-look EGDs were performed on day 1 (D1) and day 7 (D7), respectively, with EHT as needed.Post-ESD clinical bleeding rate was 2.6% (95% confidence interval [CI] 1.2%-4.9%). For clinical bleeding, adjusted odds ratios (ORs) for age <65 years and antithrombotic drug uses were 4.40 (95% CI 1.07-19.93) and 7.34 (95% CI 1.80-32.48), respectively. For D1 EHT, adjusted ORs of tumor location in the lower part of the stomach and maximum tumor diameter ≥60 mm were 2.16 (95% CI 1.35-3.51) and 2.20 (95% CI 1.05-4.98), respectively. For D7 EHT, adjusted OR of D1 EHT was 4.65 (95% CI 1.56-20.0).Post-ESD clinical bleeding rate was relatively low using 2nd and 3rd-look strategy. Age <65 years and antithrombotic drug use are significant risk factors for clinical bleeding. Regarding EHT, tumor location in the lower part of the stomach and maximum diameter of resected specimen ≥60 mm are significant predictors for D1 EHT. D1 EHT in turn is a significant risk factor for D7 EHT. The efficacy of sequential strategy for preventing post-ESD bleeding is promising.

Corrigendum: Human ß-defensin-3 inhibits migration of colon cancer cells via downregulation of metastasis-associated 1 family, member 2 expression.

In this article, Fig. 2 is incorrect. The corrected Fig. 2 is shown using data from the tissue array samples. The new figure demonstrates the same findings as the original figure. Accordingly, in the paragraph of Materials and methods, the sentence '...surgically resected colon cancer tissues' and 'This study was...research committee' and in the paragraph of Results, the sentence 'Similar results were...tissue array samples' should be deleted. The above changes do not alter the original conclusions of this study. [the original article was published in the International Journal of Oncology 45: 1059-1064, 2014 DOI: 10.3892/ijo.2014.2507].

Human ß-defensin-3 inhibits migration of colon cancer cells via downregulation of metastasis-associated 1 family, member 2 expression.

The innate immune system plays an important role as the first line of defense against many types of microbes. Accumulating reports suggest that human ß-defensins (hBDs) are expressed by and have certain roles in some cancer cells. In this study, we investigated the roles of hBD-3 in colon cancer cells. The expression of hBD-3 was examined by reverse transcriptase-polymerase chain reaction analysis of colon cancer cell lines and immunohistochemical staining of colon cancer tissues. The effect of hBD-3 on proliferation of colon cancer was assessed using the MTT assay and a real-time cell analyzer, and the effect of hBD-3 on the migration of colon cancer cells was also examined. The results showed that hBD-3 is not expressed in colon cancer cells but is produced by tumor-infiltrating monocytes. Migration of colon cancer cells was significantly inhibited by hBD-3 in a dose-dependent manner, although proliferation of colon cancer cells was not affected by administration of hBD-3. Moreover, reduced expression of metastasis-associated 1 family, member 2 (MTA2) mRNA in colon cancer cells was associated with exposure to hBD-3. In conclusion, progression of colon cancer was inhibited by hBD-3 in a paracrine fashion. Therefore, hBD-3 may be a potent new agent for treating colon cancer.

The long-term effects of splenectomy and subsequent interferon therapy in patients with HCV-related liver cirrhosis.

Partial splenic embolization (PSE) or splenectomy is widely performed to increase platelet counts for interferon (IFN) therapy. The aim of the present study was to evaluate the long-term effects of splenectomy and subsequent IFN therapy in patients with hepatitis C virus (HCV)-related liver cirrhosis (LC). The present study included 19 patients with HCV-related LC who underwent splenectomy for thrombo-cytopenia caused by hypersplenism. IFN therapy was performed in all 19 patients. The effects of splenectomy and subsequent IFN therapy on peripheral blood counts, liver function, carcinogenesis and survival rates were evaluated. Splenectomy was safely performed in all patients without major complications with the exception of portal thrombosis, which, however, it did not affect liver function when treated appropriately. Thrombocytopenia improved and IFN therapy could be performed in all the patients. A sustained virological response (SVR) was not observed in patients with genotype 1 although it was observed in 75% of patients with genotype 2. Due to severe side effects, five patients did not undergo scheduled IFN therapy. Over 5 years, the mean platelet number increased from 5.2 x 10(4) to 16.8 x 10(4)/mm3 (P<0.01) and liver function improved following splenectomy (albumin, Alb: 3.5­3.8 g/dl; total bilirubin, T-Bil: 1.0­0.7 mg/dl; prothrombin time, PT: 74.1­97.7%; total cholesterol; T-cho: 140­168 mg/dl; P<0.05). Hepatocellular carcinoma (HCC) occurred in only one patient during long­term observation and follow­up of the patients not presenting with HCC at entry. The results of the present study demonstrate that splenectomy followed by interferon therapy could be beneficial in patients with HCV-related LC.

The expression and function of Toll-like receptors 3 and 9 in human colon carcinoma.

Toll-like receptors (TLRs) are pattern-recognition receptors that are important in immune signaling. TLR recognition of various viral components including double-stranded RNA (TLR3) and unmethylated CpG-DNA (TLR9) plays a crucial role in cell survival. However, TLR expression and function in colon carcinoma cells are not well clarified. We investigated the expression of TLR3 and TLR9 in colon carcinoma cells using immunohistochemical methods. The function of TLR3 and TLR9 signaling in carcinoma cell lines was studied by direct cell stimulation with, or by cell transfection of, polyinosinic-polycytidylic acid (Poly I:C), a synthetic form of dsRNA, and by cell stimulation with CpG-oligodeoxynucleotides (ODNs), respectively. Positive TLR3 and TLR9 immunohistochemical staining was observed in 91 and 86% of human hepatocellular carcinoma (HCC) tissues, respectively. Cell surface stimulation of TLR3 with Poly I:C did not affect cell viability but it did activate NF-κB activity. By contrast, stimulation of intracellular TLRs with transfected Poly I:C significantly induced apoptosis. Cell surface stimulation of TLR9 with CpG-ODNs promoted cell proliferation, and, furthermore, these CpG-ODN TLR9 agonists reduced the cytotoxicity of the anticancer drug adriamycin. Cell surface expression of TLR3 and TLR9 in colon carcinoma cells plays an important role in cell survival. In addition, the proapoptotic activity of intracellularly expressed TLR3 may provide the possibility of using TLR3 agonists as novel clinical cytotoxic agents against colon carcinoma cells.

Sorafenib and TRAIL have synergistic effect on hepatocellular carcinoma.

A multi-kinase inhibitor, sorafenib, was recently approved and is currently recommended for the treatment of advanced hepatocellular carcinoma (HCC). However, HCC treatment outcomes are still poor and necessitate improvement. Therefore, we investigated the influence of sorafenib in combination with each of cytotoxic chemotherapy agents, hypoxia or tumor necrosis factor (TNF)-related apoptosis­inducing ligand (TRAIL), on cytotoxicity to determine which is the better adjuvant. Additive cytotoxicity of sorafenib to chemotherapy agents, hypoxia and TRAIL, to HCC cells was assessed using cell viability assay. Intracellular levels of anti-apoptotic proteins were determined using western blot analysis. Activation of Wnt/ß-catenin signaling was assessed using a luciferase reporter gene assay. Sorafenib significantly and synergistically enhanced the cytotoxicity of TRAIL to HCC cells and 4',6-diamidino-2-phenylindole (DAPI) staining showed increased apoptosis among cells treated with sorafenib and TRAIL. This augmentation in cytotoxicity was derived from sorafenib-mediated downregulation of anti-apoptotic proteins. However, sorafenib did not enhance the cytotoxicity of chemotherapy agents (cisplatin, 5-FU or doxorubicin) or hypoxic treatment to HCC. Moreover, hypoxic treatment induced Wnt/ß-catenin signaling activation. Our data showed that in combination TRAIL and sorafenib had a synergistic cytokilling effect on HCC cells and that this effect derived from sorafenib-mediated downregulation of anti-apoptotic proteins.

Clinical utility of transarterial infusion chemotherapy using cisplatin-lipiodol emulsion for unresectable hepatocellular carcinoma.

BACKGROUND: We evaluated the clinical efficacy of transarterial infusion chemotherapy using a cisplatin-lipiodol emulsion for unresectable hepatocellular carcinoma (HCC). PATIENTS AND METHODS: Fifty-seven patients with advanced HCC, with no indications for surgical resection or local ablative therapy, such as percutaneous ethanol injection and radiofrequency ablation, were enrolled in this retrospective study. RESULTS: Twelve patients were treated with cisplatin-alone at a dose of 65 mg/m(2) by infusion into the artery. Forty-two patients were treated with the same dose of cisplatin suspended in 1-10 ml of lipiodol (C/LPD). Cumulative survival rates in the cisplatin-treated group were 46.2% at one year, and 18.5% at two years, whereas these in the C/LPD group were 81.6% and 44.4%, respectively, with a significant difference between the two groups (p<0.01). In the cisplatin-treated group (n=13), no (0%) patients had a complete response (CR), two (15%) a partial response (PR), three (23%) no change (NC), and eight (62%) progressive disease (PD). In the C/LPD group (n=44), four (9%) patients had CR, 16 (35%) PR, 12 (26%) NC, and 12 (26%) PD. CR and PR were seen in 15% of the cisplatin-treated group and in 44% of the C/LPD group. C/LPD was significantly more effective than cisplatin-alone (p=0.039). Some patients showed tumor response to C/LPD after intra-arterial infusion of low-dose 5-fluorouracil. CONCLUSION: C/LPD produced superior effects compared to cisplatin-alone for unresectable HCC, causing no major side-effects, and increasing the survival rate.

Up-regulation of glypican-3 in human hepatocellular carcinoma.

AIM: To investigate the expression and significance of glypican-3 (GPC3) in human hepatocellular carcinoma (HCC). MATERIALS AND METHODS: DNA chips were used to measure the expression of mRNAs for members of the glypican and syndecan families of heparan sulfate proteoglycans (HSPGs) in normal liver tissue, non-tumor tissues and HCC. GPC3 protein expression was investigated by immunohistochemical staining in the tissues samples and Western blotting in human HCC cell lines. In addition, the levels of GPC3 protein in the blood were determined by ELISA. RESULTS: Only the expression of GPC3 was found to be markedly elevated in HCCs. In the human HCC cell lines, GPC3 expression was consistently observed, and was mainly located in the cell membrane and cytoplasm. Immunohistochemistry showed a tendency for overall staining of the cytoplasm of cells in the liver carcinoma tissues, but the cell membrane was preferentially stained in poorly differentiated HCC when compared with well-differentiated HCC. Moreover, the cell membrane was preferentially stained in metastatic lesions of HCC when compared with primary HCC lesions. Non-tumor tissues and cholangiocellular carcinoma tissues were not stained. In addition, using HepG2 cells, AG490 and piceatannol, which are signal transducer and activator of transcription 3 (STAT3) inhibitors, each increased the amount of GPC3 mRNA expressed. Assay of the circulating levels of GPC3 protein in chronic liver disease and HCC found that serum GPC3 protein levels were significantly elevated in the latter. CONCLUSION: GPC3 is highly expressed in HCC, and its expression pattern differs according to the degree of cell differentiation. In addition, the expression of GPC3 is regulated by Janus kinase-STAT signaling. GPC3 shows potential as a tumor biomarker for HCC that can be used for molecularly targeted therapy.

Functional cell surface expression of toll-like receptor 9 promotes cell proliferation and survival in human hepatocellular carcinomas.

Toll-like receptor 9 (TLR9) is a pattern-recognition receptor that is involved in immune signaling and plays a crucial role in cell survival through recognition of various bacterial and viral components including unmethylated CpG-DNA. TLR9 expression and function in cancer cells are not well understood. We investigated the expression of TLR9, and the function of TLR9 signaling, in hepatocellular carcinoma (HCC) cells following stimulation with CpG-oligodeoxynucleotides (ODNs). Positive immunohistochemical staining for TLR9 was observed in 85.7% of HCC tissues. Western blot analysis revealed that TLR9 was expressed both on the cell membrane and in the cytoplasm of HCC cell lines. Full-length TLR9 was predominantly expressed on the membrane rather than in the cytoplasm, whereas multiple cleaved forms of TLR9 were predominantly expressed in the cytoplasm rather than on the membrane. Cell surface stimulation of TLR9 promoted cell proliferation, and, furthermore, the TLR9 agonists, CpG-ODNs, reduced the cytotoxicity of the anti-cancer drug adriamycin (ADM) via up-regulation of apoptosis inhibitors such as survivin, Bcl-xL, XIAP and cFLIP, in HCC cell lines. Although cell surface stimulation of TLR9 did not activate either the NF-kappaB signaling pathway or the type-I IFN secretion pathway, gene chip microarray analysis indicated that TLR9 agonists closely regulated multiple oncology-related genes and transcription factors involved in tumorigenesis and cancer progression. In conclusion, our results indicate that functional cell surface expression of TLR9 in human HCC may play an important role in tumorigenesis and cancer progression.

Clinical significance of tumor markers in detection of recurrent hepatocellular carcinoma after radiofrequency ablation.

The aim of this study was to elucidate the importance of three tumor markers, alpha-fetoprotein (AFP), Lens culinaris agglutinin A-reactive fraction of AFP (AFP-L3), and des-gamma-carboxy prothrombin (DCP), for detecting and predicting the recurrence of hepatocellular carcinomas (HCCs). A total of 108 patients with initial non-advanced HCC who underwent curative radiofrequency ablation (RFA) in our hospital were enrolled in this study. The effectiveness of the three tumor markers for detecting recurrence and recurrence-free survival was analyzed. Positivity of these three makers was not markedly increased at the first or second recurrence. In addition, there was no significant correlation between the initial and recurrent levels of each tumor marker. The tumor marker that was positive at the time of initial HCC was not necessarily positive at recurrence. The tumor marker levels at recurrence were not correlated with pre-ablation levels. No significant correlation was found in the tumor marker values between pre-ablation and the time of recurrence. On multivariate analysis, high AFP-L3 levels (>/=10%) were significantly predictive of recurrence-free survival. All three tumor markers should be routinely measured to detect recurrence during follow-up after RFA. Especially high AFP-L3 levels should be followed closely.

Classification of hypocholesterolemia lipid patterns using Chol/Trig Combination System.

Patterns of hypocholesterolemic lipid fractions in 295 patients with liver diseases, malignant tumors, arteriosclerotic and renal diseases with cholesterol (Chol) levels of <30 mg/dl were classified using a simultaneous analytical method for the Chol and triglyceride (TG) fractions (Chol/Trig Combo System). Hypocholesterolemia was classified as follows: IV, Type IV on WHO hyperlipidemia phenotype classification; intermediate density lipoprotein (IDL), cases with appearance of IDL, including appearance of Lp(a); high + low density lipoproteins (HDL+LDL), lipids mostly consisting of HDL and LDL fractions; HDL abnormality, cases with slow alphaHDL or fast HDL; abnormal LDL, both Chol and TG fractions mostly consisting of LDL fraction; normal type, ratios of HDL, very low density lipoproteins (VLD) and LDL fractions were almost normal; and low HDL, HDL-C was <30 mg/dl. Many patients with liver diseases had HDL+LDL (45%), and abnormal LDL was noted in 13% of the cases. In malignant tumors, the frequencies of low HDL, normal type, and HDL+LDL cases were similar (22-30%). In arteriosclerosis, normal type accounted for 46% of the cases, and the frequency of normal type was higher (60%) in renal diseases. Mortality rate (within 1 year after measurement) was then compared among lipid patterns. In liver diseases, mortality rate increased in the following order: abnormal LDL (55%); low HDL (31%); HDL abnormality (25%); and HDL+LDL (21%). No deaths were seen among patients with normal type. In malignant tumors, mortality rate was very high (88%) in patients with HDL+LDL, but low in patients with normal type (22%) and low HDL (9%). Mortality rate was low in patients with arteriosclerosis and renal diseases in the short-term follow-up period (1 year). In the comparisons of distribution, mean, and appearance rate of charge modification frequency (CMF) among lipid patterns, parameters were high in all patterns other than HDL+LDL. Classification of hypocholesterolemia lipid patterns and evaluation of CMF may therefore be clinically useful.

Development of hepatocellular carcinoma in patients with chronic hepatitis C more than 10 years after sustained virological response to interferon therapy.

The risk factors for the development of hepatocellular carcinoma (HCC) in patients who have achieved a long-term sustained viral response (SVR) to interferon (IFN) are not fully understood. This study aimed to investigate the characteristics of patients who developed HCC after 10 years of achieving SVR. We retrospectively studied 5 patients with HCC which developed more than 10 years after the termination of IFN therapy. The clinical characteristics at the induction of IFN therapy were male gender, a mean age of 51.6±9.1 years, while 2 patients were moderate alcohol consumers. None of the 5 patients were positive for either HBs Ag or anti-HBc Ab. A histological examination at the initial IFN therapy showed the activity scores to be A2 in all cases, and the fibrosis scores at least F2. The clinical parameters at the diagnosis of HCC included fluctuating transaminase levels in all cases. These levels scarcely fell below the upper limits even after SVR was achieved. In 3 patients, liver tissues were obtained at the treatment of HCC. These tissues showed marked improvement in both activities and fibroses, but severe steatosis in 1 patient. To conclude, chronic hepatitis C patients who respond to IFN therapy should undergo long-term follow-up, even after the eradication of HCV, with special attention particularly to patients who had elevated transaminase levels and steatosis.