山田 宏哉

Background: Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of endothelium nitric oxide synthase (NOS). ADMA binds to a substrate-binding site of NOS and then inhibits nitric oxide production from vascular endothelial cells. Elevated ADMA levels are a risk factor for cardiovascular disease. Recently, it was reported that plasma ADMA levels were negatively correlated with vegetable and fruit consumption. The purpose of this study was to examine the association between serum levels of carotenoids and serum ADMA levels in Japanese subjects. Methods: We conducted a cross-sectional study of 470 subjects (203 men and 267 women) who attended a health examination in August 2011. Serum levels of several carotenoids were separately measured by high-performance liquid chromatography. Serum ADMA levels were determined by using an enzyme-linked immunosorbent assay kit. Results: In women, the multivariate-adjusted odds ratios (ORs) of elevated serum ADMA levels were significantly decreased in the highest tertile for beta-cryptoxanthin (OR 0.47, 95% CI 0.23-0.95), alpha-carotene (OR 0.39, 95% CI 0.18-0.79), and beta-carotene (OR 0.36, 95% CI 0.17-0.73) compared to the lowest tertile. In men, significantly decreased ORs were observed in the highest tertiles of serum zeaxanthin/lutein (OR 0.23, 95% CI 0.06-0.69) and acarotene (OR 0.26, 95% CI 0.07-0.82), and in the middle and the highest tertiles of serum beta-carotene (OR 0.27, 95% CI 0.09-0.74 and OR 0.20, 95% CI 0.03-0.88, respectively) when the tertile cutoff points of women were extrapolated to men. Conclusions: Higher serum levels of carotenoids, such as alpha-carotene and beta- carotene, may help to prevent elevated serum ADMA levels in Japanese subjects.

OBJECTIVES: An earlier study using the data from the Japanese Long-term Care Insurance (LTCI) system reported a scenario for achieving the target of Health Japan 21 (the second term): future gains in health expectancy from 2011 to 2020 must be larger than gains in expectancy. According to this scenario (the Healthy Life Expectancy Extension Scenario), the proportion of disability (cases≥Care Level 2 in LTCI disability certification) will gradually decrease by 1% per year from 2011. The purpose of this study was to estimate the cost savings in long-term care and medical care if the Healthy Life Expectancy Extension Scenario is achieved. METHODS: We used data from Japanese national statistics and a survey conducted in Osaki city, Miyagi. The natural course of disability cases (≥Care Level 2) was estimated under the assumption that the future population composition would be equal to the population projections for Japan and the future proportion of disabilities for each age grade would be equal to that of 2010. Then, the decrease in the number of disabilities based on the Healthy Life Expectancy Extension Scenario was calculated. Finally, the cost savings in long-term care and medical care associated with the assumed decrease in the number of disability cases was calculated. RESULTS: When the disability cases (≥Care Level 2) were shifted to "no disability certification (not requiring care)," a total estimated cost reduction of 5,291 billion yen was achieved from 2011 to 2020. Furthermore, a total estimated reduction of 2,491 billion yen was achieved for the same period when all disability shifts to "Care Level 1" were accounted for. CONCLUSION: As a rough calculation, if the Health Japan 21 (second term) target is achieved, approximately 2,500-5,300 billion yen will be saved in the cost of long-term care and medical care.

目的 健康日本21(第二次)の目標(平均寿命の増加分を上回る健康寿命の増加)を考慮して、健康寿命の将来予測を行った。方法 基礎資料として、国民生活基礎調査、介護給付費実態調査と簡易生命表を用いた。2011〜2020年の将来の死亡率が「日本の将来推計人口(平成24年1月推計)」のそれと同じと仮定し、将来の不健康割合に3つのシナリオを設定した。この仮定とシナリオの下で、将来の健康寿命をサリバン法で算定した。結果 「日常生活に制限のない期間の平均」では、2010年観察値(男70.4年と女73.6年)に対する2020年予測値は将来の不健康割合が2010年現在と同じというシナリオで男71.2年と女74.3年、最近の推移を継続するというシナリオで男71.4年と女74.5年、一定率で低下して将来の不健康寿命の延伸がないというシナリオで男71.7年と女74.9年であった。最後のシナリオにおける不健康割合の2010〜2020年の低下率は男0.95と女0.96と推定された。「自分が健康であると自覚している期間の平均」は2010年観察値(男69.9年と女73.3年)に対する2020年予測値が3つのシナリオで男69.5〜71.2年と女72.9〜74.6年であった。最後のシナリオにおける不健康割合の低下率は男0.96と女0.97と推定された。65歳の「日常生活動作が自立している期間の平均」は2010年観察値(男17.2年と女20.5年)に対する2020年予測値が男18.0〜18.2年と女21.2〜21.5年であった。最後のシナリオにおける不健康割合の低下率は男0.90と女0.91と推定された。結論 将来の不健康割合にシナリオを設定して、2011〜2020年の健康寿命を予測した。健康日本21(第二次)の目標が達成されるための将来の不健康割合の低下率を見積もった。(著者抄録)

目的 平成20年患者調査のオーダーメード集計に基づいて,主傷病と副傷病の関連性を検討した。方法 統計法34条に基づくオーダーメード集計を利用して,入院・外来,性・年齢階級と主傷病別の副傷病の推計患者数を得た。主傷病は傷病大分類,副傷病は糖尿病,高脂血症,高血圧(症),虚血性心疾患,脳卒中とした。入院と外来ごとに,主傷病別の副傷病の推計患者数を観察するとともに,性・年齢構成を調整した期待値に対する比(O/E比)を算定した。結果 主傷病が虚血性心疾患と脳血管疾患に対する副傷病が糖尿病,高脂血症と高血圧(症)のO/E比はいずれも1.5以上であった。O/E比が1.5以上の組み合わせとしては,主傷病が糖尿病と高血圧性疾患に対する副傷病が虚血性心疾患と脳卒中,主傷病が「糸球体疾患,腎尿細管間質性疾患及び腎不全」に対する副傷病が糖尿病,高血圧(症),虚血性心疾患と脳卒中などであった。結論 主傷病と副傷病の中に強い関連性を有する組み合わせがみられ,オーダーメード集計の利用の有用性が示唆された。(著者抄録)

Background: In many industrialized countries, non-alcoholic fatty liver disease (NAFLD) is recognized as an important disease that increases the risk of cardiovascular disease, type-2 diabetes, and metabolic syndrome. Most people with NAFLD are asymptomatic, and the disease is discovered incidentally during clinical laboratory tests. Present screening methods for NAFLD use ultrasound, and CT scans that are time-consuming, and a simple screening method for NAFLD is needed. In this study, we investigated whether serum miRNAs are associated with NAFLD and assessed the potential of serum miRNAs as a biomarker for NAFLD. Methods: We assessed intrahepatic fat by ultrasound scan, and the serum levels of five miRNAs (miR-21, miR-34a, miR-122, miR-145, and miR-451), which help regulate cholesterol and fatty acid homeostasis in liver tissue, by real-time PCR in a cross-sectional sample of 403 participants who attended health examinations. Results: Serum levels of miRNAs, miR-21, miR-34a, miR-122, and miR-451 were higher in participants with NAFLD. The serum level of miR-122 was correlated with the severity of liver steatosis. Conclusion: Serum levels of miRNAs, particularly miR-122, may be a useful biomarker for NAFLD. © 2013 Elsevier B.V.

Background: Albuminuria is a risk factor for not only nephropathy progression but also cardiovascular disease. Oxidative stress may have a role in the positive association between albuminuria and cardiovascular disease. Methods: This cross-sectional study investigated the associations of serum levels of carotenoids, which are dietary antioxidants, with albuminuria among 501 Japanese adults (198 men, mean age ± SD: 66.4 ± 10.0 years 303 women, mean age ± SD: 65.4 ± 9.8 years) who attended a health examination. Serum levels of carotenoids were determined by high-performance liquid chromatography. Logistic regression analysis was used to estimate odds ratios (ORs) with 95% CIs for albuminuria after adjustment for age, body mass index, smoking habits, drinking habits, hypertension, diabetes mellitus, and dyslipidemia. Results: Prevalence of albuminuria was 15.4% among men and 18.1% among women. Among women with albuminuria, geometric mean serum levels of canthaxanthin, lycopene, ß-carotene, total carotenes, and provitamin A were significantly lower than those of normoalbuminuric women. Adjusted ORs for albuminuria among women in the highest tertiles of serum ß-carotene (OR, 0.45 95% CI, 0.20-0.98) and provitamin A (OR, 0.45 95% CI, 0.20-0.97) were significantly lower as compared with those for women in the lowest tertile. There were no associations between serum carotenoids and albuminuria in men. Conclusions: An increased level of serum provitamin A, especially serum ß-carotene, was independently associated with lower risk of albuminuria among Japanese women. © 2013 Koji Suzuki et al.

We established a novel method to analyze cells collected by fluorescence-activated cell sorting (FACS) named mRNA quantification after FACS (FACS-mQ) in which cells are labeled with a fluorescence dye in a manner that minimizes RNA degradation, and then cells sorted by FACS are examined by analyzing their gene expression profile. In order to analyze cells using FACS-mQ, it is essential to prepare single-cell suspensions without RNA degradation. We found that a new tissue preservation medium, ThelioKeep (TM), which contains epigallocatechin-3-gallate (EGCG), was suitable for preservation of thyroid tissues. The aim of this study was to establish a cell dispersion method of thyroid follicular cells using ThelioKeep (TM). We compared the efficiency of cell dispersion between the two methods, the conventional cold pre-incubation method and the ThelioKeep (TM) method; then we determined if cells obtained by the ThelioKeep (TM) method were suitable for FACS-mQ analysis. We found that a larger number of cells were recovered using ThelioKeep (TM) than using the conventional cold pre-incubation method. Furthermore, cell viability was higher with the ThelioKeep (TM) method than with the cold pre-incubation method. Thyroid cells collected by this method were analyzed by FACS-mQ. A clear shift in flow cytometry analysis was observed when cells were stained with an anti-thyroglobulin or anti-thyroid transcription factor-1 antibody. After sorting, the same copy number of ACTB mRNA was detected in thyroid cells as in an anaplastic carcinoma cell line, 8305C. These findings imply that preparation of thyroid cells using the present method is suitable for FACS-mQ analysis.

<jats:sec><jats:title>Objective</jats:title><jats:p>3,5,3′-Triiodothyronine (T<jats:sub>3</jats:sub>)-predominant Graves' disease is characterized by the increasing volume of thyroid goiter resulting in poor prognosis. Although type 1 and type 2 iodothyronine deiodinases (<jats:italic>DIO1</jats:italic>and<jats:italic>DIO2</jats:italic>respectively) are known to be overexpressed in the thyroid tissues of T<jats:sub>3</jats:sub>-predominant Graves' disease, the pathogenesis of this disease is still unclear. The aim of our study is to identify genes that characterize T<jats:sub>3</jats:sub>-predominant Graves' disease tissue in order to clarify the molecular mechanism of this disease.</jats:p></jats:sec><jats:sec><jats:title>Design and methods</jats:title><jats:p>mRNAs from two thyroid tissues of both typical T<jats:sub>3</jats:sub>-predominant and common-type Graves' disease were analyzed with DNA microarrays with probes for 28 869 genes. Genes identified to be differentially expressed between the two groups were further analyzed in the second and third screenings using 70 Graves' thyroid tissues by real-time quantitative RT-PCR.</jats:p></jats:sec><jats:sec><jats:title>Results</jats:title><jats:p>Twenty-three candidate genes were selected as being differentially expressed in the first screening with microarrays. Among these, seven genes, leucine-rich repeat neuronal 1 (<jats:italic>LRRN1</jats:italic>), bone morphogenetic protein 8a (<jats:italic>BMP8A</jats:italic>), N-cadherin (<jats:italic>CDH2</jats:italic>), phosphodiesterase 1A (<jats:italic>PDE1A</jats:italic>), creatine kinase mitochondrial 2 (<jats:italic>CKMT2</jats:italic>), integrin beta-3 (<jats:italic>ITGB3</jats:italic>), and protein tyrosine phosphatase non-receptor type 4 (<jats:italic>PTPN4</jats:italic>), were confirmed to be differentially expressed in<jats:italic>DIO1</jats:italic>or<jats:italic>DIO2</jats:italic>over- and underexpressing Graves' tissues.</jats:p></jats:sec><jats:sec><jats:title>Conclusions</jats:title><jats:p>These genes are related to the characteristics of T<jats:sub>3</jats:sub>-predominant Graves' disease, such as high titer level of serum anti-TSH receptor antibody, high free T<jats:sub>3</jats:sub>to free thyroxine ratio, and a large goiter size. They might play a role in the pathogenesis of T<jats:sub>3</jats:sub>-predominant Graves' disease.</jats:p></jats:sec>

OBJECTIVES: We attempted to predict health expectancy in Japan for the period between 2011 and 2020, considering the target of Health Japan 21 (the second term) that future gains in health expectancy be larger than in life expectancy. METHODS: We used data from Japanese national statistics. Health expectancy between 2011 and 2020 was predicted using the Sullivan method under the assumption that future mortality was equal to the estimate in Population Projections for Japan (January 2012), and under three scenarios of future prevalence of bad health status. RESULTS: The numbers of expected years without activity limitation at birth for males and females in 2020 were predicted as 71.2 and 74.3, respectively, under the scenario that prevalence of activity limitation was constant since 2010; 71.4 and 74.5 under the scenario that the prevalence followed the recent trend; and 71.7 and 74.9 under the scenario that the prevalence decreased with such a rate that future gains in health expectancy were equal to in life expectancy. The rate of decrease in the prevalence in 2010-2020 in the last scenario was estimated to be 0.95 in males and 0.96 in females. The numbers of expected years with subjective well-being at birth in 2020 predicted under above three scenarios were between 69.5 and 71.2 in males and between 72.9 and 74.6 in females. The rate of decrease in the last scenario was estimated to be 0.96 in males and 0.97 in females. The numbers of expected years without care needs at age 65 in 2020 predicted under above three scenarios were between 18.0-18.2 in males and between 21.2-21.5 in females. The rate of decrease in the last scenario was estimated to be 0.90 in males and 0.91 in females. CONCLUSION: The health expectancy in 2011-2020 was predicted under some scenarios of future prevalence of bad health status. The rate of decrease in the future prevalence of bad health status was estimated with a view to the accomplishment of the target of Health Japan 21 (the second term).

Genome-wide association studies have revealed several susceptibility genes among patients with autoimmune thyroid disease (AITD), including CTLA4, PTPN22, FCRL3, and ZFAT. However, any possible association between these genes and AITD prognosis remains unknown. The objective of this study was to identify associations between polymorphisms of these genes and AITD prognosis. We genotyped functional polymorphisms, including CTLA4 CT60, CTLA4 +49A/G, CTLA4 -1147C/T, CTLA4 -318C/T, PTPN22 -1123C/G, PTPN22 SNP37, CD40 -1C/T, FCRL3 -169C/T, ZFAT Ex9b-SNP10, and ZFAT Ex9b-SNP2, in 197 AITD patients carefully selected from 456 registered AITD patients, and 86 control subjects. The restriction fragment length polymorphism method was used for genotyping. The CD40 -1CC genotype and C allele were significantly more frequent in patients with Graves' disease (GD) in remission than in those with intractable GD (P = 0.041 and P = 0.031, respectively). The FCRL3 -169TT genotype was significantly less frequent in patients with intractable GD than in those with GD in remission (P = 0.0324). For a ZFAT Ex9b-SNP10 polymorphism, the TT genotype and T allele were significantly more frequent in patients with severe Hashimoto's disease (HD) than in those with mild HD (P = 0.0029 and P = 0.0049, respectively). For a CTLA4 CT60 polymorphism, the antithyrotropin receptor antibody levels at the onset of GD were significantly higher in those with the GG genotype than in those with other genotypes (P = 0.0117). CD40 and FCRL3 gene polymorphisms were associated with GD intractability, and ZFAT polymorphism was associated with HD severity but not its development.

Measurement of gene expression levels in thyroid tumor cells in aspirates was difficult because it is interfered with peripheral blood cells or infiltrating lymphocytes. In this study, we established a novel method to separate thyroid tumor cells from blood cells efficiently with mesh filtration. The expression level of trefoil factor 3 (TFF3) mRNA was estimated using LGALS3 mRNA as an internal control (T/G ratio) in 148 preoperative thyroid aspirates. Intra-assay coefficients of variation (CV) of T/G ratio for high, moderate, and low samples were 6.5%, 2.5%, and 9.7%, respectively, and inter-assay CV for high, moderate, and low samples were 27.7%, 21.9%, and 38.2%, respectively. Nondiagnostic samples in terms of T/G ratio and cytology were 12.2% and 16.9%, respectively. We observed no interference with the data by contaminating blood cells. Among these patients, 12 patients received more than two repeated aspirations. We did not observe a marked day-to-day variation except in two cases. All 13 preoperative aspirates diagnosed as malignant by cytology showed an extremely low T/G ratio, whereas 93 aspirates diagnosed as benign by cytology showed extremely varied T/G ratios and 21.5% of them showed a T/G ratio below the cut-off value. Eleven cases underwent surgery. All nodules showing a low T/G ratio were diagnosed as papillary carcinoma by pathological diagnosis. However, one nodule diagnosed as follicular adenoma after surgery showed a high T/G ratio. Our present method may be a promising preoperative test for measuring mRNAs in thyroid aspirates.

Background: Although disability-free life expectancy has been investigated in Japan, gains from elimination of diseases and injuries have not been examined. Methods: We used data from the 2007 Japanese national health statistics to calculate the number of years with and without activity limitation that could be expected from eliminating 6 selected diseases and injuries. Results: At birth, the number of expected years of life without and with activity limitation was 70.8 and 8.4, respectively, in males and 74.2 and 11.8 in females. More than 1.0 expected years without activity limitation were gained from eliminating malignant neoplasms and cerebrovascular diseases; smaller gains were observed after eliminating other diseases and injuries. Elimination of cerebrovascular diseases, dementia, and fracture decreased expected years with activities of daily living (ADL) limitation, and elimination of shoulder lesions/low back pain decreased expected years with non-ADL limitation. Conclusions: Elimination of diseases and injuries increased expected years with and without activity limitation among Japanese, which suggests that improved prevention of those diseases and injuries-including cerebrovascular diseases and dementia-would result in longer disability-free life expectancy and fewer years of severe disability.

目的 平成16年国民生活基礎調査の匿名データに基づいて,健康状態と喫煙の関連を解析した。健康状態としては,自覚症状,通院状況,日常生活への影響と健康意識を取り上げた。方法 統計法36条に基づき厚生労働省から提供を受けて,匿名データを利用した。喫煙状況の得られた20歳以上の73,110人において,健康状態の調査項目ごとに,喫煙のオッズ比をロジスティック回帰により年齢を調整して算定した。結果 たばこを以前吸った者の吸わない者に対する年齢調整オッズ比は自覚症状なしを1.0とすると,36症状ともに症状ありが1.5以上であり,いずれかの症状ありが男1.62と女2.34であった。通院なしに対する年齢調整オッズ比は,13傷病の中で男の6傷病と女の12傷病の通院ありが1.5以上であり,いずれかの傷病の通院ありが男1.38と女2.10であった。日常生活の影響なしに対する年齢調整オッズ比は,日常生活の5つの活動の中で男の4活動と女の5活動の影響ありが1.5以上であり,いずれかに影響ありが男1.58と女2.42であった。健康意識がよいに対するよくないの年齢調整オッズ比は男1.57と女2.23であった。たばこを毎日吸う者と時々吸う者の吸わない者に対する年齢調整オッズ比は自覚症状,通院,日常生活の影響,健康意識ともに一定の傾向でなかった。結論 健康状態の多くの面に対して喫煙が強く関連することが確認され,匿名データ利用の有用性が示唆された。(著者抄録)

Recently, we have established an in-tube in situ hybridization method named mRNA quantification after fluorescence activated cell sorting (FACS-mQ), in which a specific RNA in a particular cell type is stained with a florescent dye, allowing the stained cells to be selected by FACS without suffering excessive RNA degradation. Using this method, the biological characteristics of the sorted cells can be determined by analyzing their gene expression profile. In this study, we used locked nucleic acid (LNA) oligonucleotides, which are known to enhance both the sensitivity and specificity of RNA detection, as hybridization probes in FACS-mQ. When we used a LNA probe targeting the human 28S sequence, we were able to efficiently separate human cells from rat cells. Using LNA probes, the hybridization step was shortened to 1 h. After the hybridization step, 84.6% RNA was preserved; thus, we were able to successfully measure gene expression levels in each type of cell after FACS. Providing the LNA probe efficiently hybridizes with the target sequence, FACS-mQ with an LNA probe is a powerful tool for separating particular cells and determining their biological characteristics by analyzing their gene expression profile.

Thyroid transcription factor-1 (TTF-1, NKX2-1) is a homeodomain-containing transcriptional factor that binds to and activates the promoters of thyroid and lung-specific genes, such as thyroglobulin, thyroid peroxidase, and thyroid stimulating hormone receptor. TTF-1 is known to play a key role in the development of the thyroid. However, the precise mechanism of TTF-1 gene transcription in human thyroid cells has not been studied. The expression of transcriptional activity in various lengths of the 5&apos;-flanking region of the human TTF-1 gene was studied in TTF-1 positive and negative human thyroid cell lines. Increased transcriptional activity was observed in thyroid cell lines containing plasmids that coded for a sequence proximal to the transcription start site of exon 1 of the TTF-1 gene. However, we did not observe any difference in promoter activity in the region up to -2.6 kb from the proximal transcription start site of the TTF-1 gene between TTF-1 positive and negative cells. These results suggest that the proximal 5&apos;-flanking region of the human TTF-1 gene does not contain sufficient cis-active regulatory information to direct gene expression in thyroid cells, and that other cis- or trans-acting factors participate in the thyroid specific gene expression of TTF-1.

Recent studies using stem cells or cancer stem cells have revealed the importance of detecting minor populations of cells in blood or tissue and analyzing their biological characteristics. The only possible method for carrying out such procedures is fluorescence-activated cell sorting (FACS). However, FACS has the following two limitations. First, cells without an appropriate cell surface marker cannot be sorted. Second, some laborious procedures such as rapid sorting or treatment under sterilized conditions may require in order to analyze their biological characteristics. If a specific mRNA in a particular cell type can be stained with a florescent dye and then the cells can be sorted without causing RNA degradation, a more simple and universal method for sorting and analyzing cells with a specific gene expression pattern could be established since the biological characteristics of the sorted cells could then be determined by analyzing their gene expression profile. In this study, we established a basic protocol for messenger RNA quantification after FACS (FACS-mQ) using a cRNA probe. This method could be used for the detection and analysis of stem cells or cancer stem cells in various tissues. (C) 2010 International Society for Advancement of Cytometry

Recent studies using stem cells or cancer stem cells have revealed the importance of detecting minor populations of cells in blood or tissue and analyzing their biological characteristics. The only possible method for carrying out such procedures is fluorescence activated cell sorting (FACS). However, FACS has the following limitations. First, cells without an appropriate cell surface marker cannot be sorted. Second, the cells have to be kept alive during the sorting process in order to analyze their biological characteristics. If an intracellular antigen that was specific to a particular cell type could be stained with a florescent dye and then the cells can be sorted without causing RNA degradation, a more simple and universal method for sorting and analyzing cells with a specific gene expression pattern could be established since the biological characteristics of the sorted cells could then be determined by analyzing their gene expression profile. In this study, we established a basic protocol for messenger RNA quantification after FACS (FACS-mQ) targeting intracellular antigens. This method can be used for the detection and analysis of stem cells or cancer stem cells in various tissues. (C) 2010 Elsevier Inc. All rights reserved.

Nestin is a maker that identifies stem cells in some adult tissues, and its expression is believed to relate to malignancy in cancer cells. In this study, we measured the expression levels of nestin mRNA in various kinds of thyroid tumor by the real-time quantitative reverse transcription-polymerase chain reaction. Unexpectedly, nestin mRNA was detected in almost all differentiated thyroid tumors and normal thyroid tissues, whereas extremely decreased expression was observed in anaplastic carcinomas, which are the most malignant of the thyroid follicular cell-derived tumors. These results suggest that nestin mRNA is a differentiation marker, and its expression does not relate to malignant characteristics in thyroid tumors. (C) 2009 Elsevier Ireland Ltd. All rights reserved.

Tensin3 is a member of tensin family which is localized in focal adhesion. In our previous study, a high level of tensin3 mRNA expression was observed in the thyroid but not ill other tissues, thus, tensin3 gene was regarded as a novel thyroid-specific gene. The high expression level of tensin3 mRNA in normal thyroid tissue suggests some fundamental roles in thyroid functions. In fact, the expression level of tensin3 mRNA was low in most thyroid carcinomas and non-functioning thyroid follicular adenomas which do [lot produce thyroid hormone. In the present Study, we measured the expression levels of tensin3 mRNA in twelve functional adenomas and compared the results with those in normal thyroid tissues, adenomatous goiters and non-functioning thyroid follicular adenomas. Tensin3 mRNA was expressed abundantly in all twelve functional adenomas at almost the same level as in normal thyroid tissues and adenomatous goiters, while its expression was significantly lower in non-functioning follicular adenomas. Considering these facts, all abundant expression of tensin3 mRNA is observed in tissues that produce thyroid hormone, Which Suggests some fundamental roles in basic thyroid functions.

Since the introduction of fine needle aspiration biopsy (FNAB) in the 1970&apos;s, a preoperative diagnostic technique for thyroid follicular carcinoma has long been awaited. Many markers that distinguish follicular carcinomas from adenomas have been reported; however, most of them have not been confirmed to be beneficial for clinical use. Trefoil factor 3 (TFF3) is a relatively new family of peptides that bears the three-loop trefoil domain. Several groups have reported that the suppression of TFF3 mRNA expression is related to malignant characteristics of thyroid follicular cell-derived tumors and the expression level of TFF3 mRNA is the most promising indicator for diagnosing follicular carcinoma. Development of TFF3-based diagnostic methods is now ongoing and it may not be long before thyroid follicular carcinoma can be diagnosed preoperatively using an aspirated sample from the tumor.

Background. The relative expression level of trefoil factor 3 (TFF3) mRNA to galectin-3 (LGALS3) mRNA (T/G ratio) is a useful marker to distinguish thyroid follicular carcinomas from adenomas. However, because of the interference by the simultaneously aspirated peripheral blood cells or infiltrating lymphocytes, the precise measurement of the T/G ratio in aspirates is difficult. Methods. We tested 2 methods of selecting thyroid tumor cells and removing blood cells from the aspirates. One method was selection with magnetic beads coated with Ber-EP4 antibody and the other was filtration through a nylon mesh. Results. After selection with Ber-EP4 antibody, T/G ratios in aspirates showed varied degrees of difference from those in the corresponding tissues. After mesh filtration, differences in T/G ratios between the aspirates and the corresponding tissues were less than 200% in all 8 cases. Conclusion. Mesh filtration of aspirates proved superior results for the measurement of T/G ratio. (c) 2008 Wiley Periodicals, Inc.

The severity of Hashimoto's disease (HD) and the intractability of Graves' disease (GD) vary among patients. To clarify whether the +869T/C polymorphism in the transforming growth factor-beta 1 (TGF-beta 1) gene, which is associated with TGF-beta 1 expression, is involved in the intractability of GD and severity of HD, we genotyped the TGF-beta 1 +869T/C polymorphism by polymerase chain reaction-restriction fragment length polymorphism method in genomic DNA samples from 33 patients with HD who developed hypothyroidism before they were 50 years old (severe HD) and 30 untreated, euthyroid patients with HD who were older than 50 years (mild HD). We also examined 48 euthyroid patients with GD who had been under treatment and were still positive for anti-thyrotropin receptor antibodies (intractable GD), 20 euthyroid patients with GD in remission and 45 healthy controls. The frequency of the T allele and the TT genotype were higher in patients with severe HD than in those with in mild HD. In contrast, the frequency of the CC genotype was higher in patients with intractable GD than in patients with GD in remission. In conclusion, the +869T/C polymorphism in the TGF-beta 1 gene is associated with the severity and intractability of autoimmune thyroid disease.