カワサキビョウ ワ イマ モ フエツズケテ イル : トクシマケンカ 10ネンカン ノ シュウケイ

Tomisaku Kawasaki saw his first case of unusual illness in a four-year-old with high fever, bilateral nonexudative conjunctivitis, redness of the lips and oral mucosa, a rash and cervical lymphadenopathy in 1961, and published in 1967. The cause of Kawasaki Disease（KD）has not known yet. In 1963, we saw a first case in Tokushima. The incidence rates per 100,000 children younger than the age of five have been steadily increasing, involving with two big prevalence in 1982 and 1986. From 1999, we enrolled 643 patients with KD for 10 years in Tokushima Prefecture. Most cases 88% were less than 5 years old and a peak incidence in children from 0 to 2 years old. KD has been more popular in winter season（1.5 times）than in other seasons. These suggest that both genetic susceptibility and environmental factors play a role in KD. In Tokushima, coronary artery aneurysm developed in 1.1%（2.4% in all Japan）. Aneurysm persist and become occlusive, thereby increasing the risk of atherosclerosis, myocardial infarction or sudden cardiac death. In 1999, the standard treatment for acute-phase KD was a 5-days regimen of intravenous（IV） gammmaglobulin（200 mg/kg/day）, supplemented with aspirin. Now, a single infusion of IV gammaglobulin（ 2 g/kg）followed by low-dose aspirin therapy. Recent regimen is more effective, but the 11% recipients were non-responder in Tokushima（20% in all Japan）. Therefor, further research is required to know the pathogenesis and host genetics in KD

Identification of Serum MicroRNAs as Novel Non-Invasive Biomarkers for Early Detection of Gastric Cancer

BACKGROUND: To investigate the potential of serum miRNAs as biomarkers for early detection of gastric cancer (GC), a population-based study was conducted in Linqu, a high-risk area of GC in China. METHODOLOGY/PRINCIPAL FINDINGS: All subjects were selected from two large cohort studies. Differential miRNAs were identified in serum pools of GC and control using TaqMan low density array, and validated in individual from 82 pairs of GC and control, and 46 pairs of dysplasia and control by real-time quantitative reverse transcription-polymerase chain reaction. The temporal trends of identified serum miRNA expression were further explored in a retrospective study on 58 GC patients who had at least one pre-GC diagnosis serum sample based on the long-term follow-up population. The miRNA profiling results demonstrated that 16 miRNAs were markedly upregulated in GC patients compared to controls. Further validation identified a panel of three serum miRNAs (miR-221, miR-744, and miR-376c) as potential biomarkers for GC detection, and receiver operating characteristic (ROC) curve-based risk assessment analysis revealed that this panel could distinguish GCs from controls with 82.4% sensitivity and 58.8% specificity. MiR-221 and miR-376c demonstrated significantly positive correlation with poor differentiation of GC, and miR-221 displayed higher level in dysplasia than in control. Furthermore, the retrospective study revealed an increasing trend of these three miRNA levels during GC development (P for trend<0.05), and this panel could classify serum samples collected up to 5 years ahead of clinical GC diagnosis with 79.3% overall accuracy. CONCLUSIONS/SIGNIFICANCE: These data suggest that serum miR-221, miR-376c and miR-744 have strong potential as novel non-invasive biomarkers for early detection of GC

Multiple-to-Multiple Relationships between MicroRNAs and Target Genes in Gastric Cancer

<div><p>MicroRNAs (miRNAs) act as transcriptional regulators and play pivotal roles in carcinogenesis. According to miRNA target databases, one miRNA may regulate many genes as its targets, while one gene may be targeted by many miRNAs. These findings indicate that relationships between miRNAs and their targets may not be one-to-one. However, many reports have described only a one-to-one, one-to-multiple or multiple-to-one relationship between miRNA and its target gene in human cancers. Thus, it is necessary to determine whether or not a combination of some miRNAs would regulate multiple targets and be involved in carcinogenesis. To find some groups of miRNAs that may synergistically regulate their targets in human gastric cancer (GC), we re-analyzed our previous miRNA expression array data and found that 50 miRNAs were up-regulated on treatment with 5-aza-2'-deoxycytidine in a GC cell line. The “TargetScan” miRNA target database predicted that some of these miRNAs have common target genes. We also referred to the GEO database for expression of these common target genes in human GCs, which might be related to gastric carcinogenesis. In this study, we analyzed two miRNA combinations, miR-224 and -452, and miR-181c and -340. Over-expression of both miRNA combinations dramatically down-regulated their target genes, <i>DPYSL2</i> and <i>KRAS</i>, and <i>KRAS</i> and <i>MECP2</i>, respectively. These miRNA combinations synergistically decreased cell proliferation upon transfection. Furthermore, we revealed that these miRNAs were down-regulated through promoter hypermethylation in GC cells. Thus, it is likely that the relationships between miRNAs and their targets are not one-to-one but multiple-to-multiple in GCs, and that these complex relationships may be related to gastric carcinogenesis.</p></div

The results of MSP analysis in primary GCs.

<p>The results of MSP analysis in primary GCs.</p

Changes of candidate target genes expression and cell proliferation after decreased DICER1 expression and/or 5-aza-CdR treatment.

<p>(<b>A</b>) The growth curves of KATO-III, HCT116 and DICER1 KO HCT116 cells. KATO-III cells were transfected with 20 nmol/l of siDICER1 or scrambled siRNA. The paired t-test was used to compare the values for the test and control samples. A value of P<0.05 was taken as significant. (<b>B</b>) Representative results of target gene expression on the RT-PCR analysis.</p

Effects of combinational transfection of miR-340 and -181c in GC cell lines.

<p>(<b>A</b>) Proliferation assaying after transfection with miR-340 and/or miR-181c mimics in KATO-III cells. (<b>B</b>) Changes in gene expression after ectopic expression of miR-340 and/or miR-181c.</p

MSP analysis of miR-224/−340 in GC, CRC cell lines and normal stomach.

<p>(<b>A</b>), (<b>C</b>) Schematic representation of the miRNAs and their host genes. Filled boxes represent the exons of the host genes and blank boxes denote the untranslated regions of the host genes. The bent arrows indicate the transcription start sites of the host genes. The vertical arrows indicate the locations of miRNAs. The vertical thick lines indicate CpG sites. Arrowheads indicate the regions examined for MSP. (<b>B</b>), (<b>D</b>) MSP analyses of miR-224 and miR-340, respectively, in GC cell lines. The bands in the ‘Mt’ lanes are PCR products obtained with methylation-specific primers, and those in the ‘Un’ lanes were obtained with unmethylation-specific primers; PBL, peripheral blood lymphocyte. The expression of miRNAs is indicated under the photographs of the MSP results.</p