A Study of Correlation between Gd-EOB-DTPA-enhanced MRI Using the 3T MRI System and Tc-99m-GSA Hepatic Scintigraphy / Hepatic Function Tests in Prehepatectomy Cases

This study compared results from Gd-EOB-DTPA on two different phases of 3T MRI with those from Tc-99m-GSA hepatic scintigraphy and hepatic function tests. Twenty-four patients with liver tumor were included in this study. All patients underwent Gd-EOB-DTPA-enhanced-MRI and Tc-99m-GSA hepatic scintigraphy. Clearance index (HH15) and receptor index (LHL15) were calculated for the Tc-99m-GSA, while signal intensities (SI) of liver at pre-injection and at 4/20min post-injection, and of spleen at 4 min/20min were measured (SIpre, SI4min, SI20min, SIsp4min, SIsp20min, respectively) for the Gd-EOB-DTPA-MRI. Liver activity at 15min by Tc-99m-GSA scintigraphy or biochemical liver function values were compared with liver spleen contrast at 4min (LSC4min = SI4min/SIsp4min) or 20min post-injection (LSC20min = SI4min/SIsp20min), and the increase in ratio at 4min (IR4min=SI4min/SIsp4min) or 20min (IR20min= SI20min/SIpre). Total bilirubin levels (T-bil), serum albumin levels (Alb), prothrombin activity, and the indocyanine green clearance test (ICG) results were also analyzed. There were statistically significant correlations in all comparisons between Gd-EOB-DTPA and Tc-99m-GSA. The highest coefficient of correlation was obtained in IR4min (LHL15: r = 0.795, P<0.001; HH15: r = -0.782, P<0.001), with IR20min (LHL15: r = 0.690, P<0.01; HH15: r = -0.528, P<0.05), LSC4min (LHL15: r = 0.458, P<0.05; HH15: r = -0.626, P<0.05), and LSC20min (LHL15: r = 0.443, P<0.05, HH15: r = -0.609, P<0.05) also significantly correlated. Correlations in hepatic function data were observed between IR4min and T-bil/Alb, and IR20min and Alb. In 3T-MRI using Gd-EOB-DTPA, the SI of liver at pre- to post-injection (especially at 4 min) significantly correlated with the corresponding Tc-99m-DTPA scintigraphy results, and with some biochemical liver function data

The Incidence of Proximal Extension of Ulcerative Proctitis in Japan and Factors Related to Proximal Extension

The incidence of proximal extension in patients with ulcerative proctitis is reported to be 18%-46%, but recent data on the incidence in Japan is inadequate. The aim of this study was to investigate the incidence of proximal extension of ulcerative proctitis and factors associated with the extension in Japan. This is a retrospective observational study involving a cohort of 53 patients with an initial diagnosis of ulcerative proctitis. Following verification of the diagnoses, demographic and clinical data were compiled. The cumulative incidence of proximal extension was estimated as ‘person-years’ and cumulative probability was calculated by the Kaplan-Meyer method. Univariate and multivariate analyses were performed to identify association factors. During a mean follow-up of 6.8 years, proximal extension was observed in 14 patients (26.4%). The cumulative incidence of proximal extension was 4.22/100 person-years and the cumulative probability at 5 years was 20.1%, consistent with recent reports from Western countries and data obtained in Japan over 2 decades ago. Univariate analysis showed active smoking (P = 0.025) and corticosteroid therapy (P = 0.006) to be risk factors in proximal extension, however multivariate analysis revealed that corticosteroid therapy was the only significant factor (P = 0.005) separating patients with and without proximal extension. No patient underwent colectomy. The incidence of proximal extension in ulcerative proctitis in Japan is comparable to that in Western countries and has not changed significantly over the past two decades. Corticosteroid therapy was identified as the only significant factor in proximal extension

Clinical Outcomes and Prognostic Factors Associated with Survival after Balloon-occluded Retrograde Transvenous Obliteration of Gastric Varices

We evaluated clinical outcomes and prognostic factors associated with survival after balloon-occluded retrograde transvenous obliteration (B-RTO) of gastric varices in patients with portal hypertension. Of 50 patients with gastric varices who underwent B-RTO, 46 (94.0%) patients in whom B-RTO was technically successful were reviewed retrospectively. Gastric and esophageal varices after B-RTO were evaluated by contrast-enhanced computer tomography and endoscopy, respectively. Liver function parameters and Child-Pugh scores were estimated before and at 1 year after B-RTO. The cumulative survival rate was calculated, and univariate and multivariate analyses were used to assess the prognostic factors. No major complications occurred in any of the patients following B-RTO and no recurrence or bleeding of gastric varices was noted. Of the 42 patients who were followed up for the progression of esophageal varices, 13 (31.0%) had worsened varices and of these, 6 (14.3%) showed bleeding. Prothrombin activity had significantly improved at 1 year after B-RTO, although there were no changes in other liver function parameters. The overall cumulative survival rates at 1, 3, and 5 years after B-RTO were 91.6%, 70.9%, and 53.6%, respectively. Multivariate analysis identified the occurrence of advanced hepatocellular carcinoma (HCC) during the observation period as a prognostic factor for survival (hazard ratio = 4.1497, 95% CI = 1.32314-13.0319, P = 0.0148). B-RTO of gastric varices is an effective treatment ensuring lower recurrence and bleeding rates; however, these patients require careful observation for progression of esophageal varices. The management of HCC is crucial for achieving long-term survival after B-RTO

Effects of Interleukin-4-Transduced Tumor Cell Vaccines and Blockade of Programmed Cell Death 1 on the Growth of Established Tumors

Interleukin (IL)-4 exhibits strong antitumor effects and IL-4 gene therapy has been used clinically in the treatment of some types of cancer. In the present study, we evaluated the efficacy of IL-4-transduced tumor cell vaccines in combination with blockade of programmed cell death 1 (PD-1) and investigated the mechanisms underlying the antitumor effects of this therapy. A poorly immunogenic murine colorectal cancer cell line (i.e. MC38) was transduced to overexpress IL-4. In a therapeutic model, MC38-IL4 cells and anti-PD-1 antagonistic antibodies (Ab) were inoculated into parental tumor-bearing mice. Immunohistochemical analyses and tumor-specific lysis were also performed. Additive antitumor effects were observed when mice were treated with IL-4 in combination with an anti-PD-1 Ab. Immunohistochemical analysis of the therapeutic model showed marked infiltration of CD4+ and CD8+ cells into established MC38 tumors of mice treated with anti-PD-1 Ab. Significant tumor-specific cytolysis was detected when the splenocytes of mice treated with both IL-4 and anti-PD-1 Ab were used as effector cells. These results suggest that blockade of the interaction between PD-1 and programmed death ligand 1 (PD-L1) enhances the antitumor immune responses induced by IL-4. Thus, IL-4 gene-transduced tumor cell vaccines in combination with PD-1 blockade may be considered as possible candidates for clinical trials of new cancer vaccines

In vivo immunological antitumor effect of OK-432-stimulated dendritic cell transfer after radiofrequency ablation

Radiofrequency ablation therapy (RFA) is a radical treatment for liver cancers and induces tumor antigen-specific immune responses. In the present study, we examined the antitumor effects of focal OK-432-stimulated dendritic cell (DC) transfer combined with RFA and analyzed the functional mechanisms involved using a murine model. C57BL/6 mice were injected subcutaneously with colon cancer cells (MC38) in their bilateral flanks. After the establishment of tumors, the subcutaneous tumor on one flank was treated using RFA, and then OK-432-stimulated DCs were injected locally. The antitumor effect of the treatment was evaluated by measuring the size of the tumor on the opposite flank, and the immunological responses were assessed using tumor-infiltrating lymphocytes, splenocytes and draining lymph nodes. Tumor growth was strongly inhibited in mice that exhibited efficient DC migration after RFA and OK-432-stimulated DC transfer, as compared to mice treated with RFA alone or treatment involving immature DC transfer. We also demonstrated that the antitumor effect of this treatment depended on both CD8-positive and CD4-positive cells. On the basis of our findings, we believe that combination therapy for metastatic liver cancer consisting of OK-432-stimulated DCs in combination with RFA can proceed to clinical trials, and it is anticipated to be markedly superior to RFA single therapy. © 2013 Springer-Verlag Berlin Heidelberg

Features of and Mechanisms Underlying Insulitis In aly/aly Male Mice as an Animal Model of Autoimmune Pancreatitis: Activation of CD11c+, CD4+, and Th2 Cells and Predominant Destruction of β-cells

Diabetes mellitus (DM) is observed in patients with autoimmune pancreatitis (AIP). The development of DM in AIP is believed to be due to blood flow obstruction of the endocrine gland that accompanies pancreatitis, as well as injury to the islets caused by inflammation. The latter is called insulitis and the detailed mechanisms underlying its development are not yet clear. The aim of the present study was to elucidate the mechanisms involved in the development of insulitis in AIP using aly mice as an animal model of AIP: results in aly/aly male mice, as the AIP group, were compared with those inaly/+ male mice as a control group. Mice in both groups were killed between 16 and 48 weeks of age, and pancreatitis and insulitis were evaluated histologically. Inflammatory and endocrine cells were evaluated by immunofluorescence staining with anti-CD4, anti-CD8, anti-CD11b, and anti-CD11c antibodies, as well as immunohistochemical analyses using insulin and glucagon antibodies. Plasma levels and the pancreatic content of interferon (IFN)-γ (as a Th1-secreted cytokine) and interleukin (IL)-4 (as a Th2-secreted cytokine) were determined. Pancreatitis was seen in aly/aly mice from 16 weeks of age and it developed gradually thereafter. Insulitis also developed gradually and was seen in mice after 24 weeks of age in association with a decrease in the number of islets. CD11c+ cells and CD4+ T cells were seen to infiltrate into the islets. Although the number of β-cells decreased with time, the number of α-cells was maintained until mice were 48 weeks of age. IFN-γ content peaked in mice at 16 weeks of age and declined rapidly from 20 weeks. There were two peaks in IL-4 content, one at 16 weeks and the other at 32 weeks, suggesting an association between IL-4 content and advanced insulitis after 32 weeks. In conclusion, the results suggest that insulitis in AIP is induced predominantly by the infiltration of CD11c+ cells and CD4+ T cells into the islets, and progression is facilitated by the imbalance of the activation of Th2 rather than Th1. Furthermore, insulitis in AIP predominantly involves β-cells rather than α-cells

CpG Island Methylator Phenotype in Primary Gastric Carcinoma

Gastric cancers (GC) with methylation of multiple CpG islands have a CpG island methylator phenotype (CIMP) and they can have different biological features. The aim of this study was to investigate the DNA methylation status of GCs and its association with their clinicopathological features. We evaluated the methylation status of four genes (MINT1, MINT2, MINT25 and MINT31) in 105 primary GCs using bisulfite-pyrosequencing analysis. We classified tumors as CIMP-high (CIMP-H), CIMP-low (CIMP-L) or CIMP-negative (CIMP-N) based on the methylation of MINT1, MINT2, MINT25, and MINT31. Overall, the prevalence of CIMP-H, CIMP-L and CIMP-N was 22% (23/105), 52% (55/105) and 26% (27/105), respectively. We observed a significant difference in tumor stage (stages I-II vs. stages III-IV) between CIMP-H and CIMP-N tumors (P = 0.0435). No significant differences were observed in clinicopathological characteristics (gender, age, location and tumor differentiation) among the CIMP phenotypes. The prognoses of patients with a CIMP-H tumor is likely to be better than those with CIMP-L or CIMP-N tumors, but these differences are not statistically significant (P = 0.074 and P = 0.200). Our results suggest that CIMP may define a subgroup of GCs with distinct biological features

DNA Methylation Profiles of Primary Colorectal Carcinoma and Matched Liver Metastasis

BACKGROUND: The contribution of DNA methylation to the metastatic process in colorectal cancers (CRCs) is unclear. METHODS: We evaluated the methylation status of 13 genes (MINT1, MINT2, MINT31, MLH1, p16, p14, TIMP3, CDH1, CDH13, THBS1, MGMT, HPP1 and ERα) by bisulfite-pyrosequencing in 79 CRCs comprising 36 CRCs without liver metastasis and 43 CRCs with liver metastasis, including 16 paired primary CRCs and liver metastasis. We also performed methylated CpG island amplification microarrays (MCAM) in three paired primary and metastatic cancers. RESULTS: Methylation of p14, TIMP3 and HPP1 in primary CRCs progressively decreased from absence to presence of liver metastasis (13.1% vs. 4.3%; 14.8% vs. 3.7%; 43.9% vs. 35.8%, respectively) (P<.05). When paired primary and metastatic tumors were compared, only MGMT methylation was significantly higher in metastatic cancers (27.4% vs. 13.4%, P = .013), and this difference was due to an increase in methylation density rather than frequency in the majority of cases. MCAM showed an average 7.4% increase in DNA methylated genes in the metastatic samples. The numbers of differentially hypermethylated genes in the liver metastases increased with increasing time between resection of the primary and resection of the liver metastasis. Bisulfite-pyrosequencing validation in 12 paired samples showed that most of these increases were not conserved, and could be explained by differences in methylation density rather than frequency. CONCLUSIONS: Most DNA methylation differences between primary CRCs and matched liver metastasis are due to random variation and an increase in DNA methylation density rather than de-novo inactivation and silencing. Thus, DNA methylation changes occur for the most part before progression to liver metastasis