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Peripheral sensory neuropathy (PSN) caused by oxaliplatin-based adjuvant chemotherapy adversely affects patients' quality of life. This study evaluated the efficacy and safety of capecitabine plus oxaliplatin (CAPOX) with intermittent oxaliplatin use compared with the standard CAPOX in adjuvant therapy for colon cancer.Patients with curative resection for stage II/III colon cancer were randomly assigned to receive either CAPOX with continuous oxaliplatin (eight cycles of CAPOX) or CAPOX with intermittent oxaliplatin (two cycles of CAPOX, four cycles of capecitabine and two cycles of CAPOX). The primary end-point was the 1-year PSN rate, and the key secondary end-point was disease-free survival (DFS).Two hundred patients were enrolled in the intent-to-treat population. After 4 patients withdrew, 196 patients were included in the safety analysis. The overall treatment completion rate was 65% for continuous vs. 89% for intermittent treatment (p 0.001). The 1-year PSN rate was 60% (95% confidence interval [CI], 50%-70%) for continuous and 16% (95% CI, 10%-25%) for intermittent treatment (p 0.001). After a median follow-up of 52 months, 40 events (20%) were observed. The 3-year DFS was 81% (95% CI, 71%-87%) for continuous and 84% (95% CI, 75%-90%) for intermittent treatment (hazard ratio [HR], 0.87; 95% CI, 0.47-1.63). Among patients with high-risk disease (T4 or N2-3), the 3-year DFS was 57% for continuous vs. 74% for intermittent treatment (HR, 0.66).CAPOX with planned intermittent oxaliplatin may be feasible as an adjuvant therapy for colon cancer and substantially reduce the duration of long-lasting PSN.UMIN000012535.

The prognosis for patients with advanced gastric cancer (GC) remains poor. The identification of biomarkers relevant to the recurrence and metastasis of GC is advantageous for stratifying patients and proposing novel molecular targets. In the present study the oncological roles of SAM domain, SH3 domain and nuclear localization signals 1 (SAMSN1), a mediator of B‑cell function, were elucidated in GC. The expression and methylation status of SAMSN1 were investigated in a panel of 11 GC cell lines. Immunohistochemical staining was performed to determine the pattern of SAMSN1 protein expression in gastric tissues. The prognostic impact of SAMSN1 expression was determined by analyzing 175 pairs of surgically resected gastric tissues. A marked decrease in the level of SAMSN1 mRNA was detected in 8/11 GC cell lines as compared with that in a non‑transformed intestinal epithelium cell line (FHs 74) without promoter methylation. The mean expression level of SAMSN1 mRNA was reduced in GC tissues compared with normal adjacent tissues, an observation that was independent of tumor differentiation. The pattern of SAMSN1 protein expression was significantly correlated with that of SAMSN1 mRNA. Low SAMSN1 mRNA expression was significantly associated with tumor size (>60 mm; P=0.026) and shorter overall survival times (P=0.004). Multivariate analysis identified low SAMSN1 mRNA expression as an independent prognostic factor for poor overall survival (hazard ratio, 1.80; 95% confidence interval, 1.07–3.05; P=0.025). The difference in survival between the low and high SAMSN1 expression groups was more marked in patients with stage II/III GC compared to those with stage IV GC. In patients with stage II/III GC who underwent curative surgery, low SAMSN1 expression was associated with reduced disease free survival times. The results of the present study indicate that downregulation of SAMSN1 transcription may affect the progression and recurrence of GC, and therefore may represent a novel biomarker of GC.

Esophageal squamous cell carcinoma (ESCC), the most common esophageal cancer in East Asia, is among the six cancers with the highest fatality rates worldwide. Unfortunately, multidisciplinary treatment strategies have not achieved satisfactory outcomes. Therefore, novel insights into the molecular biology of ESCC are required to improve treatment. The gene encoding the transmembrane adherens junctions-associated protein-1 (AJAP1) expressed by epithelial cells resides in chromosome 1p36, which is frequently lost or epigenetically silenced in several malignancies. Here, we investigated the expression levels and regulatory mechanism of AJAP1 transcription. We determined the levels of AJAP1 mRNA and the genes encoding potentially interacting proteins expressed by ESCC cell lines, as well as the chromosomal copy number of AJAP1 and the methylation status of its promoter region. AJAP1 mRNA levels of 78 pairs of surgically resected specimens were determined to evaluate the association of AJAP1 expression and clinicopathological factors. Nine ESCC cell lines differentially expressed AJAP1 mRNA, and demethylation of hypermethylated AJAP1 genomic DNA reactivated AJAP1 mRNA expression. The copy number of sequences upstream or downstream of the AJAP1 transcriptional start site was not detectably altered. AJAP1 mRNA levels correlated inversely with those of ezrin (EZR) and were significantly lower in ESCC tissues compared with adjacent normal tissues. AJAP1 mRNA levels decreased gradually with increasing tumor stage. Patients with downregulated AJAP1 transcription were more likely to experience shorter overall and disease-free survival. Multivariate analysis of disease-free survival identified downregulated AJAP1 transcription as an independent prognostic factor. These results suggest that in ESCC, AJAP1 acts as a putative tumor suppressor and that AJAP1 transcription is regulated by promoter hypermethylation. These findings indicate that downregulated AJAP1 transcription may serve as a novel tumor biomarker to predict recurrence of ESCC after esophagectomy.

Gastric cancer (GC) is a major global health problem that urgently requires novel molecular biomarkers for patient stratification as well as therapeutic targets. Anosmin-1 (ANOS1) gene encodes a cell adhesion molecule that plays diverse roles in multiple malignancies. We performed global expression profiling of GC cell lines and small interfering RNA (siRNA) experiments to determine the effect of ANOS1 expression on phenotype. We evaluated the association of ANOS1 mRNA and protein levels in patients' tissue and sera with clinicopathological factors of GC subtypes. Differential expression of ANOS1 mRNA by GC cell lines correlated positively to levels of ITGAV, FOXC2 and NODAL mRNAs and inversely with those of TFPI2. Inhibiting ANOS1 expression decreased the proliferation, invasion and migration of GC cells. The mean level of ANOS1 mRNA was significantly higher in 237 GC tissues compared with the corresponding noncancerous adjacent tissues. Elevated ANOS1 levels associated significantly with the phenotypes of GC, shorter disease-free and overall survival. ANOS1 expression was a more significant prognostic marker for diffuse and distal nondiffuse GC. ANOS1 concentrations in sera increased sequentially in sera of healthy subjects, localized GC and disseminated GCs. Prognosis was worse for patients with preoperative serum ANOS1 ≥ 600 pg/ml compared with those with

Sensitive biomarkers are necessary for risk classification of patients with gastric cancer (GC), especially ones at risk of distant metastases. Melanoma-associated antigen (MAGE)-D2 has been reported to play a role in the process of cell adhesion and metastatic potential of tumor cells in colorectal cancer. The purpose of this study was to identify a novel clinically relevant biomarker of GC. Expression analysis of MAGE-D2 was conducted in GC cell lines and clinical samples (surgical specimen and serum) in both mRNA and protein level. Correlations between MAGE-D2 expression status and clinicopathological factors were evaluated. MAGE-D2 mRNA expression levels were similar between GC tissues and the corresponding normal adjacent tissues and were independent of GC differentiation or subtype. In 101 (45 %) of 225 patients, the expression level of MAGE-D2 mRNA was increased in GC tissues compared with the corresponding normal adjacent tissues. Increased expression of MAGE-D2 mRNA in GC tissues was associated with distant metastasis and early recurrence and was an independent prognostic factor (hazard ratio 2.27, 95 % confidence interval 1.39–3.74, P = 0.001). There was a stepwise increase in serum MAGE-D2 level going from healthy volunteers to patients with localized GC and then to those with extended GC (stage IV). Patients with preoperative serum MAGE-D2 levels >130 pg/ml had a more unfavorable prognosis than those with levels ≤130 pg/ml. MAGE-D2 was associated with metastatic potential of GC and may represent a promising biomarker, both in gastric tissues and serum samples, for malignant behavior of GC.

Genetic signatures may differ by histopathologic and anatomic subtypes of gastric cancer (GC). B-cell translocation gene 1 (BTG1) was identified as one of genes downregulated in GC tissues from our microarray data. To evaluate the clinical implications of BTG1 expression in GC and the genetic diversity among GC subtypes. BTG1 mRNA expression was analyzed in GC cell lines and 233 pairs of surgical specimens. The mutational and methylation status of BTG1 in GC cell lines was analyzed, and immunohistochemistry was conducted to determine the distribution of BTG1. The pattern and prognostic significance of BTG1 expression were correlated with the three proposed GC subtypes. BTG1 mRNA was downregulated in 82 % of GC cell lines and in 88 % of clinical GC tissues. Promoter hypermethylation events or sequence mutations were not detected in GC cell lines. The pattern of BTG1 expression as observed by immunohistochemistry was consistent with that of its mRNA. Downregulation of BTG1 mRNA in GCs was significantly associated with shorter disease-specific and recurrence-free survival. Multivariate analysis of disease-specific survival identified downregulation of BTG1 transcription as an independent prognostic factor. BTG1 mRNA expression was more strongly suppressed in proximal nondiffuse and diffuse GC compared with distal nondiffuse GC, and subgroup analysis revealed that BTG1 downregulation led to adverse prognosis, specifically in patients with proximal nondiffuse and diffuse GC. Altered expression of BTG1 is a potential biomarker for carcinogenesis and progression of GC, particularly for proximal nondiffuse and diffuse GC.

Aim: Although B-cell translocation gene 1 (BTG1) plays an important role in apoptosis and negatively regulates cell proliferation, BTG1 expression in hepatocellular carcinoma (HCC) has not been evaluated. The aim of this study was to clarify the role of BTG1 in the initiation of HCC carcinogenesis and progression. Methods: BTG1 mRNA expression levels were determined for HCC cell lines and 151 surgical specimen pairs using a quantitative real-time reverse transcription polymerase chain reaction assay. The mutational and methylation statuses of HCC cell lines were analyzed via high-resolution melting analysis and direct sequencing analysis to explore the regulatory mechanisms of BTG1 expression. The expression and distribution of the BTG1 protein in liver tissues were evaluated using immunohistochemistry. Results: Decreased expression of BTG1 mRNA was confirmed in the majority of HCC cell lines (89%) and clinical HCC tissues (85%) compared with non-cancerous liver tissues. Mutations or promoter hypermethylation of BTG1 were not identified in HCC cell lines. BTG1 mRNA expression levels were not influenced by background liver status. The pattern of BTG1 protein expression was consistent with that of BTG1 mRNA. Downregulation of BTG1 mRNA in HCC was significantly associated with shorter disease-specific and recurrence-free survival rates. Multivariate analysis of disease-specific survival rates identified BTG1 mRNA downregulation as an independent prognostic factor for HCC (hazard ratio 2.12, 95% confidence interval 1.12 – 4.04, P = 0.022). Conclusions: Our results indicate that altered BTG1 expression might affect hepatocarcinogenesis and may represent a novel biomarker for HCC carcinogenesis and progression. Disclosure: All authors have declared no conflicts of interest.

Identification of novel molecules implicated in the malignancy of gastric cancer (GC) is key to the development of personalized treatments and the improvement of patient outcome. Neurotrophin receptor-interacting melanoma antigen-encoding protein (NRAGE) regulates apoptosis and metastasis via interactions with various genes. This study aimed to evaluate the function and clinical significance of NRAGE in GC. The expression of NRAGE and its putative interacting genes apoptosis antagonizing transcription factor (AATF), p75 neurotrophin receptor (p75NTR), and proliferating cell nuclear antigen (PCNA) were determined in GC cell lines using reverse transcription-polymerase chain reaction (RT-PCR). The effect of NRAGE knockdown by small interfering RNA (siRNA) on GC cell behavior also was evaluated. In addition, NRAGE expression was determined in 179 pairs of resected gastric tissues. Expression of NRAGE mRNA positively correlated with that of AATF, and NRAGE knockdown significantly decreased the proliferation, migration, and invasion of GC cells. The mean level of NRAGE mRNA expression was significantly higher in GC tissues than in corresponding adjacent normal tissues. The expression patterns of NRAGE mRNA and protein were closely correlated. A stepwise elevation in NRAGE mRNA expression in GC tissues was observed with increasing Union for International Cancer Control (UICC) stage. High NRAGE expression in GCs was associated with shortened recurrence-free survival and identified as an independent prognostic factor (hazard ratio, 1.83; 95 % CI, 1.12–3.02, p = 0.017). The results indicate that NRAGE represents a putative oncogene associated with a malignant phenotype of GC. In GC, NRAGE may serve as a predictive biomarker and a target of molecular therapy.

Hepatocellular carcinoma (HCC) is a fatal disease, primarily due to the limited effective therapies available for patients with advanced or recurrent stages of the disease. Therefore, in order to improve patient prognosis, it is important to identify an informative biomarker for HCC progression, as well as a molecular target for therapy. Neurotrophin receptor-interacting melanoma antigen-encoding protein (NRAGE), a member of the type II melanoma-associated antigen family, mediates apoptosis and cell death through interactions with a wide range of proteins, and is implicated as a tumor suppressor or oncoprotein depending on cell type. However, the role of NRAGE in HCC is currently unknown, therefore, the present study aimed to identify the underlying function of NRAGE in HCC tumorigenesis. Resected tumor and non-cancerous liver tissues from 151 patients with HCC, alongside HCC cell lines, were analyzed by polymerase chain reaction and immunohistochemical techniques to determine NRAGE expression levels, as well as the expression levels of potential genes encoding interacting proteins. It was demonstrated that the expression levels of NRAGE mRNA correlated significantly with those of apoptosis-antagonizing transcription factor (AATF), and were not affected by cirrhosis in non-cancerous liver tissues when compared to elevated levels in HCC tissues. The expression patterns of NRAGE protein and mRNA were consistent among 30 representative specimen pairs. Furthermore, increased NRAGE expression in patients with HCC correlated significantly with a shorter disease-specific survival time, and was identified as an independent prognostic factor via multivariate analysis (hazard ratio, 2.23; 95% confidence interval, 1.06-3.83; P=0.020). Therefore, the results of the present study indicated that increased NRAGE expression affects HCC progression via its interaction with AATF, and may represent a novel biomarker and molecular target for the treatment of HCC.

ササ抑制処理がヒノキ天然更新施業に及ぼす効果を検証するために, 長野県三浦実験林の帯状皆伐天然更新試験地における36年間の更新木樹冠とササの被覆の変遷を空中写真を用い経時的に解析した。1969年の伐採終了後, ササ抑制処理が行われなかった対照区では, 伐採直後を除き, ササの衰退がみられなかった。除草剤散布と刈払いによるササ抑制処理を行ったササ処理区では, ササ面積比率の低下が更新初期10年間に計3回あったと推察され, ササ抑制効果の持続期間は3年間程度であったと見積られた。2005年には, 更新木樹冠の面積比率は対照区で約24%, 処理区では70%以上であり, 1979年のササ面積比率との間に負の相関がみられた。2005年における更新林分の上層木密度は, 上層平均樹高が同等の標準的な人工林に対し, 対照区で24%, 処理区で61%に相当した。以上のことから, 天然更新の成績向上に対するササ抑制処理の寄与が示唆された。

症例は61歳の女性で，53歳時より原発性胆汁性肝硬変・シェーグレン症候群・膜性腎症にて加療中である．今回，右季肋部痛を主訴に受診し，腹部CTで肝左葉外側区域に径9 cmの腫瘍を指摘された．身体検査所見では掻痒感を認めるも黄疸はなく，血液生化学検査で胆道系酵素・IgMの上昇，抗ミトコンドリア抗体陽性を認めた．肝炎ウィルスマーカーおよび腫瘍マーカーは陰性であった．画像検査所見にて腫瘍は被膜様構造を伴う境界明瞭な腫瘤として描出された．内部は脂肪成分に富み造影による早期濃染を認めた．鑑別診断として大型の高分化型肝細胞癌と肝血管筋脂肪腫が考えられたが，確定診断はつかず肝左葉切除術を施行した．摘出標本の病理組織学的検査にて肝血管筋脂肪腫と診断された．術後1年経過した現在のところ明らかな再発所見を認めていない．今回，原発性胆汁性肝硬変の経過中に急速に増大した肝血管筋脂肪腫の1例を経験したので報告する．

はじめに：P0CY1症例の臨床病理組織学的因子，予後について検討し，P1症例と比較した．方法：当院で1996年から2005年までに手術を施行した656例の胃癌症例のうち，開腹時に腹腔洗浄細胞診（以下，CY）を施行した339例を対象とした．CYはPapanicolaou染色にて判定し，ClassVをCY1と診断した．結果：P0は281例でそのうちCY1は33例（11.7％）であった．P0CY1全例が深達度T3もしくはT4で，リンパ節転移陽性であり，P0CY0と比較してT因子，N因子の進展を認めた．生存期間を比較すると，P0CY0，P0CY1，P1の順に不良であった．腹膜播種陽性例を胃癌取扱い規約第12版に従い，P1，P2，P3に分類してP0CY1も含め比較したところ，P0CY1，P1はP3に比べ生存期間は長かった．また，P0CY1の腹腔洗浄細胞診における癌細胞数の多寡による比較を行うと，癌細胞数少数例のほうが多数例に比べ生存期間は長かった．胃切除，リンパ節郭清を行ったP0CY1のうち，有意差はないがMSTはD2群が497日，D0，D1群が264日という結果であった．考察：CY1は予後不良因子であるが，P1より生存期間は長い．腹腔洗浄細胞診における癌細胞数の多寡は予後予測因子になりうると考えられた．

Background context Spontaneous spinal epidural hematoma (SSEH) is a very rare condition, so there were few studies assessing the management criteria of SSEH. Purpose To assess the differential diagnosis and clinical results of treatment for SSEH. Study design A retrospective chart and radiograph review of the patients with SSEH. Patient sample Seven consecutive patients with SSEH who were treated in our institute. Outcome measures Differential diagnosis, severity of the paresis, and treatment selection were assessed preoperatively and postoperatively. Methods We assessed the relationship between the following parameters and clinical results: (1) the initial symptoms, (2) imaging diagnosis of magnetic resonance imaging (MRI), (3) treatment selection (conservative or surgical treatment), (4) the interval of surgery, and (5) the severity of paresis using ASIA impairment scale (AIS) grading. Results In all patients, the symptoms at onset were severe neck and back pain. MRI showed isointensity to the spinal cord in the T1-weighted view and iso- or high intensity in the T2-weighted view. A solid pattern in MRI was shown in 4 patients, and a mosaic pattern was shown in 3 patients. Decompression was performed in five cases, and spontaneous recovery appeared in two cases. The mean interval time for operation was 29.8 hours. The severity of paresis was grade B in 3 cases and grade C in 4 cases at onset. These cases recovered to become grade E in 3 cases and grade D in 4 cases. Neurological deficits were present in two patients with conservative therapy and in two patients with a long interval for operation. Conclusions Precise diagnosis without delay and rapid surgical treatment are essential for the management of SSEH.

The harmonics of a free electron laser (FEL) were irradiated in vacuum to surfaces of compressed C 60 and a mixture of C 60 and I 2 . The power and frequency of the fundamental FEL macro-pulse were ca. 0.5 mJ/pulse and 2 Hz, respectively. The irradiation time was 120–180 min. After irradiation of FEL with a typical wavelength of 450 or 345 nm, the Raman peak of Ag(2)-derived vibration mode of C 60 shifted to the lower-energy side. The Raman peak shift of the mixture powder sample was greater than that of pure C 60 . Furthermore, changes of the crystalline structure indicated that various intermolecular combinations occurred by irradiation. These results strongly suggest that three-dimensional polymerization of C 60 was promoted by laser irradiation and the effect of photon-assisted hole-doping from iodine atoms to C 60 molecules.

播種時におけるコナラ種子の乾燥が，芽生えの発達経過や成長量にどのように影響するかを調べた。播種から出芽までの期間は，無乾燥に比べ弱度の乾燥でむしろ短くなり，さらに乾燥日数が増すと長くなった。播種から出芽までの期間の長い芽生えでは上胚軸伸長期間や展葉期間は短くなった。しかし，播種から展葉終了までの全期間は，播種から出芽までの期間に依存していた。種子乾燥にともなう播種から出芽までの期間の増大は，種子の脱水率の増大で説明されなかった。種子乾燥がもたらす芽生えの成長量の低下については，播種から出芽までの期間の長期化と脱水率の増大がそれぞれ関係していた。乾燥にともなう不出芽種子の出現状況を考えあわせ，播種に際しては，乾燥日数で5 日間，脱水率では10％ に至らない取り扱いが，一応の目安になると判断した。

Gastric cancer (GC) is a major global health problem that urgently requires novel molecular biomarkers for patient stratification as well as therapeutic targets. Anosmin-1 (ANOS1) gene encodes a cell adhesion molecule that plays diverse roles in multiple malignancies. We performed global expression profiling of GC cell lines and small interfering RNA (siRNA) experiments to determine the effect of ANOS1 expression on phenotype. We evaluated the association of ANOS1 mRNA and protein levels in patients' tissue and sera with clinicopathological factors of GC subtypes. Differential expression of ANOS1 mRNA by GC cell lines correlated positively to levels of ITGAV, FOXC2 and NODAL mRNAs and inversely with those of TFPI2. Inhibiting ANOS1 expression decreased the proliferation, invasion and migration of GC cells. The mean level of ANOS1 mRNA was significantly higher in 237 GC tissues compared with the corresponding noncancerous adjacent tissues. Elevated ANOS1 levels associated significantly with the phenotypes of GC, shorter disease-free and overall survival. ANOS1 expression was a more significant prognostic marker for diffuse and distal nondiffuse GC. ANOS1 concentrations in sera increased sequentially in sera of healthy subjects, localized GC and disseminated GCs. Prognosis was worse for patients with preoperative serum ANOS1 ≥ 600 pg/ml compared with those with600 pg/ml. ANOS1 may represent a biomarker for GC phenotypes and as a target for therapy.

The prognosis for patients with advanced gastric cancer (GC) remains poor. The identification of biomarkers relevant to the recurrence and metastasis of GC is advantageous for stratifying patients and proposing novel molecular targets. In the present study the oncological roles of SAM domain, SH3 domain and nuclear localization signals 1 (SAMSN1), a mediator of B-cell function, were elucidated in GC. The expression and methylation status of SAMSN1 were investigated in a panel of 11 GC cell lines. Immunohistochemical staining was performed to determine the pattern of SAMSN1 protein expression in gastric tissues. The prognostic impact of SAMSN1 expression was determined by analyzing 175 pairs of surgically resected gastric tissues. A marked decrease in the level of SAMSN1 mRNA was detected in 8/11 GC cell lines as compared with that in a non-transformed intestinal epithelium cell line (FHs 74) without promoter methylation. The mean expression level of SAMSN1 mRNA was reduced in GC tissues compared with normal adjacent tissues, an observation that was independent of tumor differentiation. The pattern of SAMSN1 protein expression was significantly correlated with that of SAMSN1 mRNA. Low SAMSN1 mRNA expression was significantly associated with tumor size (>60 mm; P=0.026) and shorter overall survival times (P=0.004). Multivariate analysis identified low SAMSN1 mRNA expression as an independent prognostic factor for poor overall survival (hazard ratio, 1.80; 95% confidence interval, 1.07–3.05; P=0.025). The difference in survival between the low and high SAMSN1 expression groups was more marked in patients with stage II/III GC compared to those with stage IV GC. In patients with stage II/III GC who underwent curative surgery, low SAMSN1 expression was associated with reduced disease free survival times. The results of the present study indicate that downregulation of SAMSN1 transcription may affect the progression and recurrence of GC, and therefore may represent a novel biomarker of GC.

Accumulation of epigenetic alterations causes inactivation of tumor suppressors and contributes to the initiation and progression of hepatocellular carcinoma (HCC). Identification of methylated genes is necessary to improve our understanding of the pathogenesis of HCC and develop novel biomarkers and therapeutic targets. The Kallmann syndrome-1 (KAL1) gene encodes an extracellular matrix-related protein with diverse oncological functions. However, the function of KAL1 in HCC has not been examined. We investigated the methylation status of the KAL1 promoter region in HCC cell lines, and evaluated KAL1 mRNA levels and those of genes encoding potential interacting cell adhesion factors. KAL1 mRNA expression level was heterogeneous in nine HCC cell lines, and reactivation of KAL1 mRNA expression was observed in cells with promoter hypermethylation of KAL1 gene after demethylation. In addition, KAL1 mRNA levels inversely correlated with those of ezrin in all nine HCC cell lines. KAL1 expression levels in 144 pairs of surgically-resected tissues were determined and correlated to clinicopathological parameters. KAL1 mRNA level was independent of the background liver status, whereas HCC tissues showed significantly lower KAL1 mRNA levels than corresponding noncancerous liver tissues. Downregulation of KAL1 mRNA in HCC was significantly associated with malignant phenotype characteristics, including elevated tumor markers, larger tumor size, vascular invasion, and hypermethylation of KAL1. Patients with downregulation of KAL1 were more likely to have a shorter overall survival than other patients, and multivariate analysis identified downregulation of KAL1 as an independent prognostic factor (hazard ratio 2.04, 95% confidence interval 1.11-3.90, P=0.022). Our results indicated that KAL1 may act as a putative tumor suppressor in HCC and is inactivated by promoter hypermethylation. KAL1 may serve as a biomarker of malignant phenotype of HCC.

Hepatocellular carcinoma (HCC) frequently recurs after curative resection. Therefore, the availability of sensitive biomarkers for progression and recurrence is essential for managing patients’ clinical course. Adherens junctions associated protein 1 (AJAP1) may serve this purpose, because it mediates activities of tumor cells. AJAP1 mRNA levels and those of genes encoding potential interacting proteins, such as SRC in HCC cell lines, and 144 pairs of resected liver tissues were determined as well as the methylation status of the AJAP1 promoter and copy number changes at AJAP1 locus. The expression pattern of AJAP1 protein was evaluated using immunohistochemistry. AJAP1 mRNA levels varied among nine HCC cell lines, and AJAP1 expression was reactivated after demethylation of its promoter. AJAP1 mRNA levels correlated inversely with those of SRC in HCC cell lines and tissues. AJAP1 mRNA levels were suppressed in HCC tissues. The expression pattern of AJAP1 correlated significantly with that of AJAP1 mRNA. Low levels of AJAP1 mRNA in patients with HCC associated significantly with elevated levels of tumor markers, larger tumor size, serosal infiltration, vascular invasion, hypermethylation of the AJAP1 promoter, and copy number loss at AJAP1 locus. Patients with low levels of AJAP1 expression were more likely to experience shorter disease-free survival (DFS), and multivariate analysis identified low AJAP1 expression as an independent factor for predicting DFS. AJAP1 may function as a key regulatory molecule associated with the recurrence of HCC. Hypermethylation of the AJAP1 promoter is a key regulatory mechanism controlling AJAP1 expression.

Identification of molecular markers for sensitive detection of hepatocellular carcinoma (HCC) is required to achieve efficacious personalized therapy. We focused here on SAM domain, SH3 domain, and nuclear localization signals 1 (SAMSN1) and investigated expression and methylation status of SAMSN1 in HCC cell lines and 144 pairs of surgical specimens. SAMSN1 was expressed at significantly lower levels in tumor tissue compared with the corresponding noncancerous tissues of patients with HCC. Analysis of HCC cell lines revealed that hypermethylation of the SAMSN1 promoter correlated with decreased expression of SAMSN1 mRNA. Furthermore, treating cells with a DNA-demethylating drug increased SAMSN1 transcription. The levels of SAMSN1 mRNA in noncancerous liver were not affected by background liver inflammation or fibrosis. Moreover, the levels of SAMSN1 mRNA in HCC tissues inversely correlated with tumor size and preoperative levels of proteins induced by vitamin K absence. The clinical significance of SAMSN1 was further indicated by the correlation between its decreased expression in patients with HCC and their shorter overall and recurrence-free survival as well as recurrence following initial resection. Moreover, multivariate analysis identified SAMSN1 as an independent prognostic factor of HCC progression. The expression pattern of SAMSN1 correlated significantly with that of SAMSN1 mRNA, making it possible to use PCR techniques to readily quantitate SAMSN1 expression in tumors. Our findings indicate that inhibition of SAMSN1 transcription through DNA hypermethylation may influence the progression of HCC and thus represent a novel biomarker of the phenotype of HCC cells.

Seedlings of the three coniferous species including Thujopsis dolabrata var. hondai Makino were raised under weak shade in a nursery in order to examine the photochemical and nonphotochemical dissipations of excited energy at PSII. The dry matter growth 20 weeks after germination was smallest in T. dolabrata var. hondai. This was due to the low photosynthetic capacity in addition to the small leaf weight ratio of the seedlings and the large leaf specific weight. The specific ranking of rETRmax (the maximum relative electron transport rate) was different from that of NPQmax (the maximum of non-photochemical quenching) ; T. dolabrata var. hondai was characterized by the high NPQmax relative to the low rETRmax. A slight depression in Fv/Fm, (light stress parameter) was recognized only for Cryptomeria japonica in the cotyledon period. A large depression in Fv/Fm was observed for C. japonica and T. dolabrata var. hondai in the post-cotyledon period. These depressions in Fv/Fm seemed to be closely associated with the low levels of rETRmax but may have also been related to the effects of NPQmax and foliage morphology and architecture. The light responsibility of T. dolabrata var. hondai seedlings differed greatly from that of Pinus densiflora Sieb. et Zucc. and differed significantly from that of C. japonica.

Microsatellite variations in Castanopsis species in Japan were examined to clarify the genetic relationships among 25 local populations according to the difference in the number of layers of adaxial epidermis in the leaves. Six microsatellite loci were assayed for 629 seedlings from the populations, and these seedlings were classified into five types according to the state of the leaf epidermis. Remarkable differences in the allele frequency of the six microsatellite loci were observed among these local populations. The coefficients of genetic differentiation, R(ST), of each locus ranged from 0.209 to 0.388. An unweighted pair-group method (UPGMA) phenogram constructed on the population pairwise R(ST) over the loci revealed three clusters (A-C), and six sub-clusters. These clusters reflected the differences in the occurrence frequency of seedlings in each epidermis type within a population. Our findings suggest that clusters A and C are the local populations dominated by Castanopsis sieboldii and Castanopsis cuspidata, respectively, while local populations of cluster B are composed of the two Castanopsis species and/or include many individuals derived by hybridization. The six sub-clusters were found to reflect the geographic relationship among the populations, suggesting a different process for geographic population dynamics during the postglacial period.

上層木を強くすかした林地にコナラ果実を播き付け, 落葉被覆が実生の発生と成長に及ぼす影響を調べた。また, 圃場で，落葉被覆が地表下3 cmの土壌の温度とpF 値に及ぼす影響を調べた。落葉被覆の影響は，土壌のpF 値に対して顕著であった。被覆量200 g m-2 以下では土壌乾燥は急速に進んだ。一方，400 g m-2 以上では土壌乾燥は大きく抑制された。実生の発生率は，被覆量90 g m-2 では5% であったが，被覆量180 g m-2 および360 g m-2 ではともに40% を示した。1 成長期を経た実生重は，被覆量によって異なり，360 g m-2 では180 g m-2に比べ20% 近く大きかった。実生重については，果実重や実生発生時期のほかに，シュート伸長様式が関与しており，落葉被覆は実生発生時期と二次伸長の促進にかかわっていた。厚い落葉被覆は，実生発生時期を遅らせるが，二次伸長を促進した。厚い落葉被覆がもたらす土壌の湿潤性が，二次伸長の促進を通して，実生の成長量を増加させると考えた。

Identification of novel genetic and epigenetic alterations is required for optimal stratification of patients with hepatocellular carcinoma (HCC) at risk for recurrence and adverse prognosis. Coenzyme Q10 (CoQ10), which mediates apoptosis, is synthesized by prenyl diphosphate synthase subunit 2 (PDSS2). In the present study we evaluated the clinical significance and regulatory mechanisms of PDSS2 expression in HCC. PDSS2 expression levels and those of genes encoding potentially interacting proteins as well as the methylation status of the PDSS2 promoter region were analyzed in HCC cell lines. PDSS2 mRNA levels in 151 pairs of resected specimens were determined to evaluate the association of PDSS2 expression and clinicopathological factors. The expression and distribution of PDSS2 were determined using immunohistochemistry. PDSS2 mRNA expression was decreased in six of nine HCC cell lines and significantly correlated with those of hepatocyte nuclear factor 4α. PDSS2 transcription in HCC cells with decreased PDSS2 expression accompanying hypermethylation was reactivated after treating these cells with a methylation inhibitor. Mean expression levels of PDSS2 mRNA relative to that of uninvolved liver diminished gradually in the order of chronic hepatitis to cirrhosis, and each was significantly higher than those of HCCs. PDSS2 and PDSS2 mRNA levels were consistent. Decreased PDSS2 mRNA levels were detected in HCC tissues of 56 patients, correlated with shorter disease-specific survival, and was identified as an independent prognostic factor. PDSS2 is a putative tumor suppressor, and promoter hypermethylation is a key regulatory mechanism in HCC. Decreased levels of PDSS2 mRNA expression may represent a novel biomarker of HCC.

Background Gastric cancer (GC) remains to have a poor prognosis via diverse process of cancer progression. Dihydropyrimidinase-like 3 (DPYSL3) is a cell adhesion molecule that has been reported to be involved in the metastatic process of tumor cells. The aim of this study was to identify a novel clinically-relevant biomarker of GC. Methods Expression analysis of DPYSL3 mRNA and protein levels was conducted using GC cell lines and 238 pairs of surgically resected gastric tissues. Correlations between expression status of DPYSL3 and clinicopathological parameters were investigated. Results DPYSL3 mRNA expression levels positively correlated with those of potentially interacting genes (VEGF, FAK and EZR) in GC cell lines. GC tissues from tumors with distant metastases (stage IV cancer) showed elevated expression levels of DPYSL3 mRNA. The DPYSL3 staining intensity in immunochemical staining was consistent with the mRNA expression patterns in GC tissues. High DPYSL3 mRNA expression in GCs was significantly associated with more malignant phenotypes and was an independent prognostic factor. Moreover, patients with high DPYSL3 mRNA expression had a significantly shorter recurrence free survival after curative resection. In subgroup analysis based on tumor histology, similar tendency was observed between patients with differentiated and undifferentiated GCs. Conclusions Expression status of DPYSL3 in GC tissues may represent a promising biomarker for the malignant behavior of GC.

Patients with hepatocellular carcinoma (HCC) may relapse after curative resection. Sensitive biomarkers for HCC are required to enhance disease management. Dihydropyrimidinase-like 3 (DPYSL3) suppresses cell proliferation and tumorigenicity of certain malignancies; however, its role in HCC is unknown. The expression levels of DPYSL3 and genes encoding potential interacting proteins vascular endothelial growth factor (VEGF), focal adhesion kinase (FAK), ezrin, and cellular src were determined using RT-PCR. Further, we determined the methylation status of the DPYSL3 promoter in HCC cells lines and the effect of inhibiting DPYSL3 expression on their phenotype. DPYSL3 expression was determined in 151 pairs of resected liver tissues. DPYSL3 mRNA levels were down-regulated in most HCC cell lines with DPYSL3 promoter hypermethylation, and expression was restored after demethylation. DPYSL3 expression levels inversely correlated with those of VEGF and FAK. Knockdown of DPYSL3 significantly increased migration and the invasive properties of HCC cells. The mean level of DPYSL3 mRNA was significantly lower in HCC tissues compared with corresponding noncancerous tissues. The expression patterns of DPYSL3 mRNA and protein were consistent. DPYSL3 mRNA expression in HCC tissues inversely correlated with preoperative serum tumor markers and was significantly lower in patients with extrahepatic recurrences. Disease-specific and recurrence-free survival was significantly shorter in patients with down-regulated DPYSL3 expression. Our results indicate that DPYSL3 is a putative HCC tumor suppressor, and promoter hypermethylation potently regulates DPYSL3 transcription. Down-regulation of DPYSL3 expression in HCC tissues may serve as a predictive biomarker for HCC after curative resection.

Hepatocellular carcinoma (HCC) is the most common cause of cancer-related mortality globally. Since the prognosis of advanced HCC patients is extremely poor, the development of novel molecular targets for diagnosis and therapy is urgently required. In the present study, the expression of the melanoma-associated antigen-D2 (MAGE-D2) gene was investigated to determine whether it affects the malignant phenotype of HCC and thus, may serve as a marker of prognosis. Therefore, the expression of MAGE-D2 mRNA and MAGE-D2 protein in nine HCC cell lines and 151 pairs of surgical tissues was analyzed. mRNA expression levels were analyzed using reverse transcription-quantitative polymerase chain reaction and immunohistochemistry was used to compare the clinicopathological parameters of the tumors. A significant difference in the level of MAGE-D2 expression was observed between the normal liver and chronic hepatitis tissues, however, no significant differences were identified among the levels of the chronic hepatitis, cirrhosis and HCC tissues. The expression patterns of the MAGE-D2 protein were consistent with those of its mRNA. The expression levels of MAGE-D2 mRNA in 66 of 151 (44%) patients were higher in the HCC tissues compared with the corresponding non-cancerous tissues. In addition, the disease-specific survival time was significantly shorter for patients with higher levels of MAGE-D2 mRNA expression. Multivariate analysis identified increased expression of MAGE-D2 mRNA as an independent prognostic factor for disease-specific survival (hazard ratio, 2.65; 95% confidence interval, 1.43-4.98; P=0.002). However, increased expression levels of MAGE-D2 mRNA were not significantly associated with other clinicopathological parameters, including extrahepatic recurrence. These results indicated that MAGE-D2 mRNA affects tumor progression and may serve as a prognostic indicator following curative resection. In addition, MAGE-D2 may provide a target for the therapy of HCC.

Patients with advanced gastric cancer (GC) have an adverse prognosis even after curative resection. Development of novel diagnostic and therapeutic approaches for GC is urgently required. The expression and methylation status of DENN/MADD domain-containing protein 2D (DENND2D), a member of the membrane trafficking proteins, were evaluated in 12 GC cell lines and 112 pairs of surgical specimens. Subgroup analysis based on tumor differentiation, location, and morphology was also performed. Expression and distribution of DENND2D protein were determined by immunohistochemistry. The majority of GC cell lines (75 %) and tissues (79 %) showed reduced expression of DENND2D mRNA compared with noncancerous gastric tissues. GC tissues showed a significantly lower mean expression level of mRNA and a higher frequency of promoter hypermethylation of DENND2D than corresponding noncancerous tissues. No significant differences in DENND2D mRNA expression and methylation status were found between GC subtypes categorized by tumor differentiation, location, and morphology. The expression patterns of DENND2D protein were confirmed to be consistent with those of DENND2D mRNA. Downregulation of DENND2D mRNA in GC tissues was significantly associated with factors related to more advanced GC and subsequent adverse prognosis. Among 72 patients who underwent R0 resection, downregulation of DENND2D mRNA in GC tissues was an independent prognostic factor and associated with early recurrence. Our results suggested that DENND2D is a putative tumor suppressor gene regulated by promoter hypermethylation in GC. Downregulation of DENND2D can serve as a novel tumor biomarker to predict progression and early recurrence of all types of GC.

Seasonal courses of leaf CO2 gas exchange in a growing season were examined in saplings ofThujopsis dolabrata var.hondai andQuercus mongolica var.grosseserrata in a cool temperate deciduous forest. Between the two tree species there were no large differences in the light compensation point of leaf photosynthesis, except for the season of new leaf expansion. However, light-saturated rates of net photosynthesis were obviously high inT. dolabrata var.hondai. EvergreenT. dolabrata var.hondai saplings had large photosynthetic production in two seasons, before the emergence of new foliage and after foliage fall of the overstory deciduous trees, because of the significantly high solar radiant energy penetrating under the forest canopy during the seasons. Saplings of deciduousQ. mongolica var.grosseserrata were heavily shaded throughout the growing season by foliage of the overstory trees, which resulted in a low daily surplus production. The annual surplus production of leaves in the growing season was estimated to be 2300 mmol CO2 m−2 inT. dolabrata var.hondai and −100 mmol CO2 m−2, slightly negative, inQ. mongolica var.grosseserrata. These results supported the high survivability ofT. dolabrata var.hondai saplings and the high mortality ofQ. mongolica var.grosseserrata in the deciduous forest.

The rectangular hyperbola, non-rectangular hyperbola, and monomolecular function as empirical models of photosynthetic light-response curve, were applied to data acquired from saplings of two Quercus species growing in the shaded undergrowth of a deciduous secondary-forest. The rectangular hyperbola gave unacceptably high estimates of the initial slope (φ) and the upper asymptotic maximum (Pmax) . The monomolecular function gave the best estimates for Pmax. However, the non-rectangular hyperbola was considered to be more reliable in estimating values of φ. In the three parameters estimated by the non-rectangular hyperbola, the interspecific difference in Pmax was almost negligible. As for θ, a curvature factor (dimensionless), Q. mongolica var. grosseserrata showed higher values although limited to a growth season and statistically nonsignificant. It was the parameter φ that the interspecific difference was significant, where Q. serrata showed higher values. From the estimation that the higher values of parameter φ act more effectively on the leaf carbon gain in heavily shaded forest floors, the saplings of Q. serrata were understood to be more shade-tolerant than Q. mongolica var. grosseserrata.

林冠木の伐採による急激な光環境の変化がコナラ前生稚樹の成長反応に及ぼす影響を明らかにするため, 光環境の異なる4試験区 (林内区, 大小の林冠ギャップ区および林外区) に, 成長期前の4月と成長期途中の7月にポットに移植した前生稚樹を配置し, 葉群形成や葉の光合成光反応曲線について調べた.4月に配置した稚樹では, 大ギャップ区と林外区のもので二次伸長が起こり, それぞれ一次葉の1.5および2倍の二次葉が形成された.一次葉の光飽和光合成速度 (Pmax) は, 林外区を除けぼ, 相対日射量の上昇にともない増大したが, 光反応曲線の初期勾配 (∅) の変化は小さかった.林外区の一次葉の∅ とPmaxは, ギャップ区のそれに比べ低く, 強光阻害を受けていた.二次葉のPmaxは高く, 大ギャップ区では一次葉の1.4倍, 林外区では2.2倍の値を示した.7月にギャップ区に配置した稚樹では, 二次伸長は起こらず, Pmaxの増大も小さかった.また, 林外区の稚樹では, 一次葉の脱落とそれに続く新葉の再生が起こった.稚樹の成長量は, 相対日射量の上昇にともない増大し, そこでは光合成能力の高い二次葉や再生葉の出現が新たな光環境下での高い生産性に大きく関与していた.

コナラ二次林の林冠下に生育する稚樹のCO2交換について, 4年生個体を対象として調べ, 枯死にいたる外的および内的要因を分析した。林冠下において葉の光合成生産を高めるうえで重要な光合成光反応曲線の初期勾配は, 成長期の大部分を通じて高いレベルにあり, 季節によるちがいは小さかった。成長期における個体の葉の年総光合成量と呼吸量および非同化器官の呼吸量は, 1993年と1994年の平均で, それぞれ29.7, 17.8, 12.5×103μmol CO2 seedling-1と推定され, CO2収支はほぼ均衡していた。成長期における個体の炭素収支は, 林床日射量の高い成長期の初めに蓄積した余剰生産物をそれ以降で消費する形で推移し, とくに夏場では高温による呼吸の増大で収支が破綻しやすいと見られた。葉の純生産率とC/F比との関係が検討され, 年齢を経た個体ではC/F比の10から20%の上昇により葉の純生産率が半分以下にまで低下することが示された。試算された生存限界C/F比は5付近にあり, 稚樹群の枯死過程を議論するうえで有用であった。

Photosynthetic properties of leaves are compared among saplings of tree species. The interspecific difference is insignificant in maximal rate of net photosynthesis (Pmax) but is recognized in initial slope of light response curve (x) . Values of α are largest in C. controversa and smaller in M. kobus and Q. mongolica var. grosseserrata: P. verecunda and Q. serrata have intermediate values. Under exposure to an irradiance of 1, 400μmol quanta m-2 sec-1, the photosynthetic response (P*) declines rapidly for Q. serrata and Q. mongolica, whereas the other tree species show no rapid decreases at least for 5 min, also followed by relatively gentle decreases. The photosynthetic reduction of Pmax by the 60-min exposure to high light ranges from 16% of M. kobus to 35% of P. verecunda and that of a from 28% of Q. mongolica to 55% of P. verecunda, significantly differing due to species. However, the extent of photosynthetic reduction of Pmax or α is not directly correlated with the decrease in P*. It is saplings of C. controversa that is presumably able to utilize most efficiently both weak and strong light under the actual light regime of the forest floor. Saplings of Q. serrata and Q. mongolica, by contrast, have no particularly advantageous photosynthetic properties to either use of weak light or that of strong light.

The VLSI design of OFDM baseband transceiver is described, which is dedicated to cognitive radio systems. To facilitate dynamic spectrum accessing, this receiver architecture employs a specialized FFT module and a mapping/demapping module. Efficient mechanisms to omit input/output of unused subrcarriers are introduced. Also, calculation of spectrum intensity for subcarrier level carrier sensing is successfully integrated to the FFT module. Whole transmitter/receiver modules are implemented by maintaining compatibility with IEEE 802.11g. Implementation results show that the proposed architecture can process OFDM symbols in 60 MHz operation.

VLSI architecture for OFDM baseband transceiver is developed, which is dedicated to cognitive radio systems. To facilitate dynamic spectrum access, this transceiver architecture employs a specialized FFT/IFFT module and a mapping/demapping module. Efficient mechanisms to omit input/ output of unused subcarriers are introduced. Also, calculation of spectrum intensity for subcarrier level carrier sensing is successfully integrated to the FFT module. Whole transmitter/ receiver modules are implemented by keeping compatibility with IEEE 802.11g. A prototype system is constructed by using Xilinx Virtex-II FPGA, which can transmit video sequence in real time.

A 17-year-old male suffered recurrent subarachnoid hemorrhages due to multiple aneurysms in the distal branches of the cerebral arteries. Autopsy revealed arteriopathy as well as the aneurysms. The arteriopathy was widespread, affecting the distal branches of small and medium-sized muscular cerebral arteries, as well as the anterior and posterior spinal arteries, and vasocorona. The arteriopathy was characterized by prominent intimal thickening, discontinuity or absence of the internal elastic lamina, and thinning and/or disappearance of smooth muscle in the tunica media with fibroplasia. Slight intimal thickening was also observed in the arteries of the circle of Willis and its major branches, as well as in the basilar artery. However, the arterioles, venules and veins showed no remarkable features. The arterial lesions were found only in the central nervous system. The multiple aneurysms in the distal branches of the cerebral arteries, which had produced the main symptoms and clinical signs, were due to the arteriopathy.

The light compensation point for CO2 gas exchange is formulated as a function of 2 physiological factors, dark respiration and initial slope of the photosynthetic response curve to light intensity. The response surface curve of light compensation point given by the above model explains how the 2 physiological factors concern the change in light compensation point with temperature or the inter-specific difference. The increase in light compensation point for young growths of T. dolabyata var. hondae with increasing temperature is insignificant until the optimum temperature of photosynthesis. This is due to that initial slope of the photosynthetic response curve increases in parallel with the raise in dark respiration. In the higher temperature region the remarkable increase in light compensation point is seen mainly due to the increase in dark respiration. Light compensation points of young growths of T. dolabyata var. hondae are in the range of 4 to 6 μmol quanta m-2 sec-1 at the optimum temperature. Thus they do not have particularly lower compensation points but rather higher values compared with other woody species growing on the same forest floor. Lower light compensation points of the seed-lings of Q, mongolica var. grosseserrata and A. japonica var. borealis is attributed to the low respiration, and those of C. harringtonia var. nana to the effects of the low respiration and steep initial slope of the photosynthetic response curve.

A novel approach to hardware implementation of H.264 block size decision is proposed, which is based on rate-distortion (RD) optimization. Utilization of RD cost for block size decision can improve up to 2.0 dB of PSNR in compared with conventional SAD/SATD based approaches. However, calculation of RD cost for a block incurs considerable computational costs since distortion can be determined only after completing the whole encoding processes of the block. Thus the proposed approach simplifies VLC process and our hardware employs 7 stage pipeline architecture for the cost calculation. As a result, the proposed architecture, which can be implemented by 20k gates, achieves real-time processing of SD (720times480) frames at a rate of 30 fps in 23.7 MHz operation

Aim: Patients with hepatocellular carcinoma (HCC) have a poor prognosis, and novel molecular targets for treating recurrence and progression of the disease along with associated biomarkers are urgently required. In the present study, expression and the regulatory mechanism of TUSC1 (tumor suppressor candidate 1) were investigated to determine if it is a candidate tumor suppressor gene for HCC, which shows repressed transcription that involves aberrant DNA methylation. Methods: TUSC1 mRNA expression levels in HCC cell lines and 94 pairs of surgical specimens were determined using quantitative real-time reverse transcription polymerase chain reaction assay. Methylation status of HCC cell lines and clinical samples were analyzed to investigate the regulatory mechanism of TUSC1 transcription and the relationship between the methylation status of the TUSC1 gene and clinicopathological factors. The expression and distribution of the TUSC1 protein in liver tissues were determined using immunohistochemistry. Results: A majority of HCC cell lines (89%) and surgical specimens (84%) demonstrated reduced expression levels of TUSC1 mRNA compared with paired non-cancerous liver tissues. The mean mRNA expression level in HCC was significantly lower than in corresponding non-cancerous liver. In contrast, no significant difference was found in TUSC1 mRNA expression level between adjacent normal and cirrhotic liver tissue from HCC patients. The TUSC1 protein expression pattern in HCC and liver tissues was consistent with TUSC1 mRNA expression. Twenty-nine (31%) of 94 patients showed intragenic hypermethylation of the TUSC1 gene in HCC, and hypermethylation was significantly associated with advanced pathological stage. Subsequently, patients with hypermethylation of TUSC1 gene had a significantly poorer prognosis than patients without hypermethylation. Conclusions: Our results suggest that TUSC1 is a candidate tumor suppressor gene and intragenic hypermethylation is one of the suppressive mechanisms that regulate TUSC1 transcription in HCC. Intragenic methylation of the TUSC1 gene may serve as a novel prognostic marker of HCC. Disclosure: All authors have declared no conflicts of interest.

In young sugi (Cryptomeria japonica D. Don) stands, crown shape (crown length/crown diameter) ratio, average branch inclination, and spatial density of foliage in the crown increased with stand age. Within crowns, foliage distribution increased from the apex downward and, until crown closure, reached a maximum near the crown base. After crown closure, the maximum occurred near the middle of the crown. In each stand, foliage distribution in the canopy showed almost the same vertical change over time as it did in individual crowns. The vertical distribution of foliage in the canopy moved upward with stand age, accompanied by an increase in canopy depth and leaf mass. The shape of the vertical distribution was almost symmetrical between the upper and lower halves in the closed stands, although slightly skewed downward. The logarithm of average spatial density decreased linearly as cumulative leaf mass increased with distance from the top of the canopy. The total cross-sectional area of the crowns exceeded the stand area from the middle of the canopy downward in the closed stands because of crown overlap. However, partly because of changes in crown morphology and structure, the increase in leaf mass with stand age did not always cause more severe crown competition.