山田 宏哉

OBJECTIVE: Dietary intake of vegetables is one of the key lifestyle factors associated with preventing cancer and cardiovascular disease (CVD). Although previous studies have provided evidence that dietary factors can alter global DNA methylation levels in humans, little work has been done on dietary factors influencing methylation levels of specific genes associated with CVD. The aim of this study was to examine whether dietary intake of vegetables was associated with adenosine triphosphate-binding membrane cassette transporter A1 (ABCA1) DNA methylation levels in leukocytes in a Japanese population. METHODS: This cross-sectional study included 279 Japanese adults (125 men, 154 women) without any clinical history of cancer, stroke, or ischemic heart disease. ABCA1 DNA methylation levels in leukocytes were measured using a pyrosequencing method. Information on dietary vegetable intake was obtained from the validated food frequency questionnaire. RESULTS: Mean ABCA1 DNA methylation levels in men and women were 35.6% ± 6.5% and 36.9% ± 6.7%, respectively. In women, multivariable linear regression analysis showed that the group with the highest dietary vegetable intake (carrot, broccoli, pumpkin, and all vegetables) showed significantly lower levels of ABCA1 DNA methylation than the lowest intake group (P = 0.04, <0.001, 0.001, and 0.02, respectively). No significant association was observed between dietary intake of vegetables and DNA methylation levels in men. CONCLUSIONS: High dietary intake of vegetables was associated with decreased ABCA1 DNA methylation levels in Japanese women. This may contribute to a better understanding of the protective effects of dietary vegetable intake on CVD.

Taking action in response to health examination results is important to stay healthy. We aimed to investigate the associations between occupation, employment type and company size, and having a health examination and taking action in response to the results among Japanese employees. We focused on three particular actions by employees in response to health examination results: paying attention to one's health, receiving health guidance, and visiting a medical institution. We used anonymous data from the 2010 Comprehensive Survey of Living Conditions of Japan, a self-administered nationwide questionnaire survey. The data of 23,963 employees (12,938 male and 11,025 female) aged 20-64 yr were analyzed using logistic regression models adjusted by covariates. There were significant changes in odds ratios for receiving a health examination by occupation, employment type and company size. We found significant odds ratios for receiving health guidance by occupation and company size, but there was almost no significant association with paying attention to one's health and visiting a medical institution. These results confirmed that receiving a health examination was associated with occupational factors, and suggested that receiving health guidance after health examination results was associated with occupation and company size.

© 2019 Elsevier B.V. Objectives: MicroRNAs (miRNAs) dysregulate gene expression by binding to target messenger RNAs, and play an important role in the pathogenesis of various diseases, including cancers, cardiovascular diseases and diabetes. Circulating miRNAs have increasingly been recognized as biomarkers for detecting and diagnosing those diseases. Few studies have investigated the association of circulating miRNA with the early stages of cognitive impairment, such as mild cognitive impairment, in the general population. The purpose of this study was to examine the association between cognitive function and several serum miRNAs levels related to amyloid precursor protein (APP) proteolysis in a Japanese general population who had never been diagnosed with dementia. Methods: We conducted a cross-sectional study of 337 Japanese subjects (144 men, 193 women) who attended a health examination. The short form of the Mini-Mental State Examination (SMMSE) was used to assess cognitive function. Serum levels of 6 miRNAs (let-7d, miR-17, miR-20a, miR-27a, miR-34a, miR-103a) were measured by quantitative real-time polymerase chain reaction. Results: Multivariable-adjusted odds ratios (ORs) for lower SMMSE score (SMMSE score < 28) were significantly increased in the lowest tertile of serum miR-20a (OR, 2.08; 95% confidence interval (CI), 1.09–4.04) and miR-103a (OR, 1.91; 95%CI, 1.00–3.69) compared to the highest tertile. Moreover, serum levels of miR-20a, -27a, and -103a were linearly and positively associated with SMMSE scores after adjustment for confounding factors. Conclusion: Low serum levels of miR-20a, -27a, and -103a are independently associated with cognitive impairment.

Global fructose consumption is on the rise; however, maternal high-fructose intake may have adverse effects on offspring. We previously demonstrated that excessive fructose intake by rat dams altered steroidogenic gene transcription in the hippocampus of offspring. Herein, we examined how maternal high-fructose intake influences the regulation of adrenal glucocorticoid levels in offspring. Rat dams received 20% fructose solution during gestation and lactation. After weaning, the offspring were provided normal water. Maternal high-fructose intake did not alter mRNA expression levels of adrenal corticosterone-synthesizing and corticosterone-inactivating proteins or the circulating adrenocorticotropic hormone levels of offspring at postnatal day (PD) 21; however, it increased circulating corticosterone levels and decreased mRNA and protein levels of adrenal 5α-reductase type 1 and 11β-hydroxysteroid dehydrogenase type 2 in offspring at PD160. Furthermore, maternal high-fructose intake enhanced DNA methylation of the adrenal 5α-reductase 1 promoter region in PD160 offspring. Thus, maternal high-fructose intake was found to affect adrenal steroid hormone clearance in adult offspring - at least in part - through epigenetic mechanisms.

AIM: Aberrant global DNA methylation is involved in the development of several diseases, including cardiovascular disease (CVD). We investigated whether the methylation of long interspersed nuclear element-1 (LINE-1) in leukocytes is associated with dyslipidemia, a major risk factor for CVD, in the Japanese general population. METHODS: We conducted a cross-sectional study consisting of 420 Japanese subjects (187 men and 233 women) without a clinical history of cancer, stroke, or ischemic heart disease. LINE-1 DNA methylation levels in leukocytes were measured using a pyrosequencing method. RESULTS: Significantly higher odds ratios (ORs) for hypermethylation were observed in the high LDL cholesterol and high LDL/HDL ratio groups than the corresponding normal group (high LDLC group: OR, 1.88; 95% confidence interval [CI], 1.20-2.96, high LDL/HDL ratio group: OR, 1.90; 95% CI, 1.20-3.01). Subjects with 2 or more lipid abnormalities had significantly higher ORs for hypermethylation than those with no lipid abnormality (OR, 2.31; 95% CI, 1.11-4.82). CONCLUSION: LINE-1 DNA hypermethylation in leukocytes was associated with CVD risk profiles: high LDLC, high LDL/HDL ratio, and the degree of abnormal lipid metabolism.

BACKGROUND: Most studies of plasma adiponectin (APN) and mortality among community-dwelling elderly focus on cardiovascular disease, but data on the relationship between plasma APN and cancer mortality is exiguous. We investigated whether APN is associated with cancer mortality in community-dwelling elderly people. METHODS: We conducted a case-cohort study within the New Integrated Suburban Seniority Investigation (NISSIN) Project using a randomly drawn sub-cohort of 697 subjects (351 men and 346 women; mean age 64.5 [standard deviation, 0.5] years) among whom we compared cases of all-cause death (n = 269) and cancer death (n = 149) during a mean follow-up duration of 10.8 (standard deviation, 3.7) years. Associations between APN and mortality were assessed using weighted Cox regression analyses. RESULTS: We observed significant positive associations between the APN concentration and cancer death in the first and third APN tertiles compared with the second APN tertile (hazard ratio [HR]T1 vs T2, 1.67; 95% confidence interval [CI], 1.00-2.79 and HRT3 vs T2, 2.10; 95% CI, 1.30-3.40). Further adjustment for possible confounders attenuated the association (HRT1 vs T2, 1.63; 95% CI, 0.93-2.84 and HRT3 vs T2, 2.10; 95% CI, 1.26-3.50). A similar but weaker association was seen for all-cause mortality (multivariate HRT1 vs T2, 1.45; 95% CI, 0.95-2.21 and HRT3 vs T2, 1.51; 95% CI, 1.01-2.25). CONCLUSION: Plasma APN and cancer mortality have a significant relationship among community-dwelling elderly people, which warrants further study.

AIMS: Recent increases in fructose consumption have raised concerns regarding the potential adverse intergenerational effects, as maternal fructose intake may induce physiological dysfunction in offspring. However, no reports are available regarding the effect of excess maternal fructose on reproductive tissues such as the ovary. Notably, the maternal intrauterine environment has been demonstrated to affect ovarian development in the subsequent generation. Given the fructose is transferred to the fetus, excess fructose consumption may affect offspring ovarian development. As ovarian development and its function is maintained by 17β-estradiol, we therefore investigated whether excess maternal fructose intake influences offspring ovarian estradiol synthesis. Rats received a 20% fructose solution during gestation and lactation. After weaning, offspring ovaries were isolated. KEY FINDINGS: Offspring from fructose-fed dams showed reduced StAR and P450(17α) mRNA levels, along with decreased protein expression levels. Conversely, attenuated P450arom protein level was found in the absence of mRNA expression alteration. Consistent with these phenomena, decreased circulating levels of estradiol were observed. Furthermore, estrogen receptor α (ERα) protein levels were also down-regulated. In accordance, the mRNA for progesterone receptor, a transcriptional target of ERα, was decreased. These results suggest that maternal fructose might alter ovarian physiology in the subsequent generation.

microRNAs (miRNAs) play important roles in regulation of gene expression during cervical carcinogenesis. We investigated expression profiles of miRNAs in cervical cancer and its precursor lesions by utilizing cervical mucus. Cervical mucus was collected from 230 patients with a normal cervix, cervical intraepithelial neoplasia (CIN), squamous cell carcinoma (SCC), or adenocarcinoma (AD). The levels of miRNA in the mucus were quantified by miRNA array and real-time reverse-transcriptase polymerase chain reaction (RT-PCR). The performance for detecting diseases was statistically analysed. The expression of miRNAs was further validated in the surgical tissues of enrolled patients. Four miRNAs (miR-126-3p, -20b-5p, -451a, and -144-3p) were significantly up-regulated in SCC and AD compared with normal, and their expression levels correlated with disease severity and high-risk human papillomavirus infection. Receiver operating characteristic curve analyses revealed that the area under the curve values for miR-126-3p, -20b-5p, -451a, and -144-3p were 0.89, 0.90, 0.94, and 0.93, respectively, for SCC plus AD compared with normal, showing high accuracy of cancer detection. Real-time RT-PCR analyses confirmed the expression of these four miRNAs in frozen tissues from cervical cancer. miR-126-3p, -20b-5p, -451a, and -144-3p in cervical mucus are promising biomarkers for cervical cancer and high-grade CINs.

Recent increases in fructose consumption have raised concerns about the potential adverse intergenerational effects of excess fructose intake. In the present study, we investigated whether excess maternal fructose intake affects hippocampal function in offspring. Female Sprague-Dawley rats were divided into 3 experimental groups: one group received distilled water, one group received 20% fructose water, and one group received 20% glucose water in addition to standard chow during gestation and lactation. Hippocampal function of offspring was evaluated by using novel object recognition and fear conditioning tests. Impaired cognitive performance was observed in the offspring of fructose-fed dams at postnatal d 60, potentially a result of decreased hippocampal neurogenesis. Real-time PCR analysis demonstrated that offspring from fructose-fed dams exhibited decreased brain-derived neurotrophic factor ( BDNF) gene expression, whereas pyrosequencing assays revealed increased DNA methylation at the BDNF promoter. The potential association between BDNF transcription and levels of DNA methylation was confirmed on the basis of luciferase activity. Furthermore, longitudinal analysis revealed that increased methylation of the BDNF promoter region was maintained at least until rats reached maturity. These results indicate that epigenetic changes associated with BDNF may underlie hippocampal dysfunction that is induced by early-life exposure to excess maternal fructose consumption.-Yamazaki, M., Yamada, H., Munetsuna, E., Ishikawa, H., Mizuno, G., Mukuda, T., Mouri, A., Nabeshima, T., Saito, K., Suzuki, K., Hashimoto, S., Ohashi, K. Excess maternal fructose consumption impairs hippocampal function in offspring via epigenetic modification of BDNF promoter.

Background: Recently, several studies have shown that microRNAs are present in high-density lipoprotein, and high-density lipoprotein-microRNA may be a promising disease biomarker. We investigated the stability of high-density lipoprotein-microRNAs in different storage conditions as this is an important issue for its application to the field of clinical research. Methods: microRNAs were extracted from the high-density lipoprotein fraction that was purified from the serum. miR-135 a and miR-223, which are known to be present in high-density lipoprotein, were quantified by quantitative real-time PCR. The influence of preanalytical parameters on the analysis of high-density lipoprotein-miRNAs was examined by the effect of RNase, storage conditions, and freezing and thawing. Results: The concentrations of microRNA in high-density lipoprotein were not altered by RNase A treatment (0-100 U/mL). No significant change in these microRNAs was observed after storing serum at room temperature or 4 degrees C for 0-24 h, and there was a similar result in the cryopreservation for up to two weeks. Also, high-density lipoprotein-microRNAs were stable for, at least, up to five freeze-thaw cycles. Conclusions: These results demonstrated that high-density lipoprotein-microRNAs are relatively resistant to various storage conditions. This study provides new and important information on the stability of high-density lipoprotein-microRNAs.

Neurosteroids, steroidal hormones synthesized de novo from cholesterol within the brain, stimulate hippocampal functions such as neuron protection and synapse formation. Previously, we examined the effect of maternal fructose on the transcriptional regulation of neurosteroidogenic enzymes. We found that the mRNA expression level of the steroidogenic acute regulatory protein (StAR), peripheral benzodiazepine receptor (PBR), cytochrome P450(11), 11-hydroxysteroid dehydrogenase (HSD), and 17-HSD was altered. However, we could not determine whether maternal fructose intake played a role in the gestation or lactation period because the dam rats were fed fructose solution during both periods. Thus, in this study, we analyzed the hippocampi of the offspring of dams fed fructose during the gestation or lactation period. Maternal fructose consumption during either the gestation or lactation period did not affect the mRNA levels of StAR, P450(17), 11-HSD-2, and 17-HSD-1. PBR expression was down-regulated, even when rats consumed fructose during the lactation period only, while fructose consumption during gestation tended to activate the expression of P450(11)-2. We found that maternal fructose intake during gestation and lactation differentially affected the expression of hippocampal neurosteroidogenic enzymes in the offspring.

Background: Understanding the roles of circulating microRNAs (miRNAs) can provide important and novel information regarding disease pathogenesis and a patient's clinical condition. Circulating miRNAs, such as exosomal miRNA, may regulate various bioactivities related to intercellular communication. However, the circulation of miRNAs in Graves' disease (GD) in relation to disease activity has never been elucidated. This study aimed to identify circulating miRNAs in GD in relation to disease activity and whether their exosomes play a role in the pathogenesis of GD. Methods: Circulating miRNAs were measured in serum obtained from seven intractable GD patients, seven GD patients in remission, and seven healthy controls using the miScript miRNA PCR Array. Altered miRNAs selected from array data were validated in 65 subjects. To investigate exosome biology, peripheral blood mononuclear cells (PBMCs) were incubated with exosomes isolated from the subjects' sera. mRNAs were quantified for cytokines using quantitative real-time polymerase chain reaction. Results: Circulating miR-23b-5p and miR-92a-39 were increased in GD patients in remission compared with intractable GD patients (p&lt;0.05). On the other hand, let-7g-3p and miR-339-5p were decreased in GD patients in remission compared with intractable GD patients (p&lt;0.05). Exosomes from intractable GD patients stimulated mRNA expression for IL-1 and TNF- compared with GD patients in remission or healthy controls. Conclusions: This study demonstrates that different levels of circulating miRNAs are associated with intractable GD. Moreover, serum exosomes of patients with intractable GD may activate immune cells, which may play an important role in GD pathogenesis.

Aims: Fructose may play a crucial role in the pathogenesis of metabolic syndrome (MetS). However, the pathogenic mechanism of the fructose-induced MetS has not yet been investigated fully. Recently, several reports have investigated the association between mitochondrial DNA (mtDNA) and MetS. We examined the effect of fructose-rich diets on mtDNA content, transcription, and epigenetic changes. Main methods: Four-week-old male Sprague-Dawley rats were offered a 20% fructose solution for 14 weeks. We quantified mRNAs for hepatic mitochondrial genes and analyzed the mtDNA methylation (5-mC and 5-hmC) levels using ELISA kits. Key findings: Histological analysis revealed non-alcoholic fatty liver disease (NAFLD) in fructose-fed rats. Hepatic mtDNA content and transcription were higher in fructose-fed rats than in the control group. Global hypomethylation of mtDNA was also observed in fructose-fed rats. Significance: We showed that fructose consumption stimulates hepatic mtDNA-encoded gene expression. This phenomenon might be due to epigenetic changes in mtDNA. Fructose-induced mitochondrial epigenetic changes appear to be a novel mechanism underlying the pathology of MetS and NAFLD. (C) 2016 Elsevier Inc. All rights reserved.

Autoimmune thyroid diseases (AITDs), including Graves' diseases (GD) and Hashimoto's thyroiditis (HT), are the most common autoimmune diseases, and are mainly mediated by T cells that produce cytokines and chemokines in abnormal amounts. Few reports have described the circulating chemokines active in AITDs. Recently, we used a new multiplex immunobead assay to simultaneously measure cytolcines and chemokines in small volume serum samples from patients with AITDs. We measured 23 selected serum chemokines in patients with GD (n=45) or HT (n=26), and healthy controls (n=9). GD patients were further classified as either untreated, intractable, or in remission, while HT patients were classified as either hypothyroid or euthyroid. Of the 23 serum chemokines assayed, only the serum level of IP-10 (CXCL10/interferon-gamma-inducible protein 10) was elevated, depending on disease activity, in GD or HT compared with healthy controls. However, the serum level of IP-10 was also increased in both untreated GD patients and hypothyroid HT patients, suggesting that levels of this cytokine may not be affected by disease specificity. In conclusion, autoimmune inflammation in patients with AITD is closely related to the level of the serum chemokine, IP-10. Therefore, IP-10 might be a good biomarker for tissue inflammation in the thyroid, but not a useful biomarker for predicting disease specific activity, the progression of AITDs, or responsiveness to treatment because of its independence from thyroid function or disease specificity.

It has been unclear whether the prevalence of disability is higher in an area affected by natural disaster than in other areas even if more than one year has passed since the disaster. The aim of this ecological study was to examine whether the rate of increase in disability prevalence among the older population was higher in disaster-stricken areas during the 3 years after the Great East Japan Earthquake (GEJE) and tsunami. This analysis used public Long-term Care Insurance (LTCI) data covering 1570 municipalities. "Disaster areas" were considered to be the three prefectures most affected by the earthquake and tsunami: Iwate, Miyagi, and Fukushima. The outcome measure was the number of aged people (&gt;= 65 years) with LTCI disability certification. Rates of change in disability prevalence from January 2011 to January 2014 were used as the primary outcome variable, and compared by analysis of covariance between "coastal disaster areas", "inland disaster areas" and "non-disaster areas". The mean rate of increase in disability prevalence in coastal (14.7%) and inland (10.0%) disaster areas was higher than in non-disaster areas (6.2%) (P &lt; 0.001). During the 3 years after the earthquake, the increase of disability prevalence from before the GEJE continued to be higher in the disaster-stricken areas. (C) 2015 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

目的 国民生活基礎調査の世帯数と患者調査の推計患者数について、東日本大震災によって調査対象から除外された地域の補完を行った。方法 国民生活基礎調査において、東日本大震災によって調査対象から除外された2011年の岩手県・宮城県・福島県と2012年の福島県の世帯数について、前後の大規模調査年の情報を用いて線型内挿法で補完した。2011年における宮城県と福島県の推計患者数について、同年の患者調査(宮城県の石巻・気仙沼医療圏と福島県が調査対象から除外)と2012年の福島県患者調査、2011年と2012年の医療施設調査と病院報告を用いて補完した。結果 全国の世帯数(各年6月時点)における補完値は2011年で48,732千世帯、2012年で48,874千世帯であった。世帯構造別の世帯数において、2011年と2012年の調査値が前後の年次よりも大きく落ち込んでいたのに対して、両年の補完値には落ち込みがなかった。2011年10月の推計患者数の補完値は、全国で入院1,365.4千人と外来7,383.9千人、施設所在地が宮城県で入院21.2千人と外来130.0千人、施設所在地が福島県で入院22.0千人と外来108.8千人であった。調査値に対する補完値の比を性・年齢階級と傷病分類ごとにみると、全国の推計患者数ではほぼ1.02倍であったが、患者住所地が宮城県と福島県の推計患者数ではきわめて大きかった。結論 国民生活基礎調査の世帯数と患者調査の推計患者数の補完値を示した。年次推移の観察にあたって、補完の仮定を十分に考慮することが重要であろう。(著者抄録)

人工心肺などの体外循環により、活性化した免疫細胞である白血球が炎症性サイトカインを分泌し、炎症カスケードを増幅していく。この機構を実験的に調べるため、ヒトリンパ系株化細胞U937を用いて増殖効果と、循環液中のサイトカイン(Interleukin1-β、Tumor Necrosis Factor-α:TNAα)および細胞中のmRNA発現を測定し、その影響を検討した。これは体外循環の影響を単一細胞で調べ、臨床検討の一助とすることを目的にしている。この研究の結果、細胞を循環させることで細胞の増殖時期が早まること、サイトカイン産生量に変化はないが、TNAαのmRNA発現がやや増加することなどが示された。(著者抄録)

Background: Circulating microRNAs (miRs) may be promising biomarkers for several diseases. We previously found that miR-122 can function as a biomarker for non-alcoholic fatty liver disease (NAFLD). However, little is known regarding the time course of circulating miR-122 levels during the development of NAFLD. Here, we examined circulating miR-122 levels using a rat model of NAFLD. Methods: To clarify changes in serum levels of miR-122 during development of NAFLD, experimental rats were fed a high-fat diet (HFD) for 2-10 weeks, while control rats received standard chow. Serum and liver tissue was collected from all animals at 2, 6, and 10 weeks of feeding. Clinical laboratory parameters (cholesterol, TG, AST, ALT, NEFA) were determined by biochemistry analyzer. Hepatic lipid accumulation was estimated by Oil red 0 staining. Circulating miR-122 levels were then measured by real-time polymerase chain reaction. Results: Over the 10 weeks of feeding, body weight, total liver lipids, and liver and serum triacylglycerol were increased in the HFD group compared to the control group. However, no significant changes in serum alanine aminotransferase activity were observed, suggesting that NAFLD status was mild. In contrast, we observed drastic up-regulation of circulating miR-122 levels. Our findings suggest that serum miR-122 level is indeed useful for assessing early NAFLD and might be superior to clinical markers traditionally used to monitor hepatic disease. (C) 2015 Elsevier B.V. All rights reserved.

Objectives: We examined whether selected factors were associated with activity limitation used to calculate the healthy life expectancy in accordance with the target of Health Japan 21 (the second term). Methods: Data for 6251 subjects were obtained from the Comprehensive Survey of Living Conditions and the National Health and Nutrition Survey, both of which were conducted by the Ministry of Health, Labour and Welfare of Japan in 2010. The age-adjusted odds ratios (AOR) of limitation of activity for the assessed factors were estimated using a logistic model.Results:, The percentage of persons with activity limitation was 12.1% of men and 15.6% of women. For men, low body mass index (BMI) (AOR: 2.02, p=0.008), high blood pressure (AOR: 1.53, p=0.021), high hemoglobin A1c (HbA1c) (AOR: 1.99, p=0.000), a small number of steps (AOR: 1.68, p=0.002), and high intake of salt (AOR: 0.69, p=0.010) were significantly associated with limitation of activity. For women, high BMI (AOR: 1.49, p=0.003), a small number of steps (AOR: 1.48, p=0.009), and high intake of salt (AOR: 0.77, p=0.017) were significantly associated with activity limitation. Conclusion: We identified several factors that were associated with activity limitation. Our results from cross-sectional data require careful interpretation before concluding whether these relationships are causal

Background: The evidence for an association between low intake of vegetables and fruits and increased colorectal cancer risk is inconclusive. Evaluating the colorectal cancer risk associated with continued low intake is important. Methods: We used data of 45 516 and 14 549 subjects aged 40-79 years obtained in the baseline and interim surveys, respectively, from the Japan Collaborative Cohort Study for Evaluation of Cancer Risk (JACC Study). The intake frequency of vegetables and fruits as assessed by a self-administered questionnaire was classified into tertiles of low, middle, and high groups, and the low group was subdivided into 2 equal groups (lower low and higher low groups). Colorectal cancer incidence determined from follow-up was used. Cox's proportional hazard model was employed to estimate the hazard ratios (HRs) and 95% confidence intervals (CIs), adjusting for covariates. Results: During 598 605 person-years of subject follow-up after baseline, we identified 806 colorectal cancer cases. HRs for the lower low versus the middle and high intake frequencies of vegetables and fruits at baseline were 0.95 (95% CI 0.77-1.16) and 1.08 (95% CI 0.90-1.29), respectively. During 125 980 person-years of subject follow-up after the interim survey, 197 colorectal cancer cases were identified. HRs for the low versus middle and high intake frequencies of vegetables and fruits in both baseline and interim surveys were 0.91 (95% CI 0.61-1.37) and 0.87 (95% CI 0.59-1.27), respectively. Conclusions: Our results suggest that low intake and continued low intake of vegetables and fruits are not strongly associated with colorectal cancer risk.

Background: Epidemiologic studies have reported coffee consumption to be associated with various health conditions. The purpose of this study was to examine the relationship of coffee consumption with colorectal cancer incidence in a large-scale prospective cohort study in Japan. Methods: We used data from the Japan Collaborative Cohort Study for Evaluation of Cancer Risk (JACC Study). Here, we analyzed a total of 58 221 persons (23 607 men, 34 614 women) followed from 1988 to the end of 2009. During 738 669 person-years of follow-up for the analysis of colorectal cancer risk with coffee consumption at baseline, we identified 687 cases of colon cancer (355 males and 332 females) and 314 cases of rectal cancer (202 males and 112 females). We used the Cox proportional-hazard regression model to estimate hazard ratio (HR). Results: Compared to those who consumed less than 1 cup of coffee per day, men who consumed 2-3 cups of coffee per day had an HR of 1.26 (95% confidence interval [CI] 0.93-1.70), and men who consumed more than 4 cups of coffee per day had an HR of 1.79 (95% CI 1.01-3.18). A statistically significant increase in the risk of colon cancer was associated with increasing coffee consumption among men (P for trend -0.03). On the other hand, coffee consumption in women was not associated with incident risk of colon cancer. Coffee consumption was also not associated with rectal cancer incidence in men or women. Conclusions: This large-scale population-based cohort study showed that coffee consumption increases the risk of colon cancer among Japanese men.

Context Autoimmune thyroid diseases (AITDs), including Graves' disease (GD) and Hashimoto's thyroiditis (HT), are the most common autoimmune diseases. MicroRNAs (miRNAs) are small noncoding RNAs, which can play pivotal roles in immune functions and development of autoimmunity. Recently, it has been recognized that identification of circulating miRNAs can provide important and novel information regarding disease pathogenesis and clinical condition. However, the role circulating miRNAs in AITD has not yet been described. Objective The aim of this study was to characterize the different circulating levels of miRNA in patients with AITD. Design and methods Sixty-four participants who met the criteria for HT or GD and healthy subjects were recruited. Microarrays were used to analyse the expression patterns of miRNA in serum obtained from patients with HT and GD and healthy subjects. After analysing the microarray data, four interesting miRNAs (miR-16, miR-22, miR-375 and miR-451) were selected and validated by quantitative real-time PCR. Results Several miRNAs were observed to be differently expressed in serum from patients with AITD compared with healthy subjects by microarray analysis. Further analysis consistently showed that serum levels of miR-22, miR-375 and miR-451 were increased in patients with HT. On the other hand, the serum levels of miR-16, miR-22, miR-375 and miR-451 were increased in patients with GD compared with healthy subjects. Conclusions We revealed that different levels of serum miRNAs were associated with GD and HT, which may play a role in the pathogenesis of these diseases.