Effects of tachyplesin on the morphology and ultrastructure of human gastric carcinoma cell line BGC-823

AIM To investigate the morphological and ultrastructural changes in the human gastric carcinoma cell line BGC-S23 after being treated with tachyplesin, METHODS Tachyplesin was isolated from acid extracts of Chinese horseshoe crab (Tachypleus tridentatus) hemocytes, BGC-823 cells and the cells treated with 2.0 mg/L tachyplesin were examined respectively under light microscope, scanning and transmission electron microscope. RESULTS BGC-823 cells had undergone the restorational alteration in morphology and ultrastructure after tachyplesin treatment. The changes were as follows: the shape of cells was unanimous, the volume enlarged and cells turned to be flat and spread, the nucleocytoplasmic ratio lessened and nuclear shape became rather regular, the number of nucleolus reduced and its volume lessened, heterchromatin decreased while euchromatin increased in nucleus. In the cytoplasm, mitochondria grew in number with consistent structure relatively, Golgi complex turned to be typical and well-developed, rough endoplasmic reticulum increased and polyribosome decreased. The microvilli at cellular surface were rare and the filopodia reduced while lamellipodia increased at the cell edge. CONCLUSION Tachyplesin could alter the malignant morphological and ultrastructural characteristics of human gastric carcinoma cells effectively and have a certain inducing differentiation effect on human gastric carcinoma cells

Effects of tachyplesin on the regulation of cell cycle in human hepatocarcinoma SMMC-7721 cells

AIM: To investigate the effects of tachyplesin on the cell cycle regulation in human hepatcarcinoma cells. METHODS: Effects of tachyplesin on the cell cycle in human hepatocarcinoma SMMC-7721 cells were assayed with flow cytometry. The protein levels of p53, p16, cyclin D1 and CDK4 were assayed by immunocytochemistry. The mRNA levels of p21(WAF1/CIP1) and c-myc genes were examined with in situ hybridization assay. RESULTS: After tachyplesin treatment, the cell cycle arrested at G(0)/G(1) phase, the protein levels of mutant p53, cyclin D1 and CDK4 and the mRNA level of c-myc gene a were decreased, whereas the levels of p16 protein and p21(WAF1/CIP1) mRNA increased. CONCLUSION: Tachyplesin might arrest the cell at G(0)/G(1) phase by upregulating the levels of p16 protein and p21(WAF1/CIP1) mRNA and downregulating the levels of mutant p53, cyclin D1 and CDK4 proteins and c-myc mRNA, and induce the differentiation of human hepatocacinoma cells

Effects of tachyplesin on proliferation and differentiation of human hepatocellular carcinoma SMMC-7721 cells

AIM: To investigate the antitumor activities of tachyplesin on human hepatocellular carcinoma (HCC) cells. METHODS: Tachyplesin, isolated from acid extracts of Chinese horseshoe crab ( Tachypleus tridentatus) hemocytes, was used to treat the human HCC cell line SMMC-7721. Effects of tachyplesin on the proliferation of SMMC-7721 cells were measured with trypan blue dye exclusion test and HE staining. The morphology and ultrastructure of the cells were examined by light microscopy and transmission electron microscopy, respectively. The activities of gamma-glutamyltransferase (gamma-GT) and tyrosine aminotransferase (TAT) were assayed with biochemical methods. The levels of alpha fetoprotein (alpha-FP), proliferating cell nuclear antigen (PCNA), p21(WAF1/CIP1) and c-myc were examined by immunocytochemistry. RESULTS: After treatment with tachyplesin 3.0 mg/L, the proliferation of SMMC-7721 cells was inhibited significantly, with the cell growth inhibitory rate amounted to 55.57% and the maximum cell mitotic index declined by 43.68%. The morphology and ultrastructure underwent restorational alteration. The activity of gamma-GT declined while TAT activity increased obviously, and the levels of alpha-FP and PCNA decreased. Moreover, the expression of p21(WAF1/CIP1) protein was up-regulated and that of c-myc protein was down-regulated. CONCLUSION: Tachyplesin could effectively inhibit the proliferation of hepatocarcinoma cells, reverse the malignant morphological and ultrastructural characteristics, alter the levels of enzymes and antigens, regulate the expression of differentiation-associated oncogene and tumor suppressor gene, and induce hepatocarcinama cell differentiation

電子支払決済法制の新潮流(OECDモバイル・オンライン決済に関する消費者政策ガイダンス / EU決済サービス指令2015（PSD2) / 大韓民国電子金融取引法2016改正と支払決済)

　科学技術の著しい発展に伴う金融手法の開拓に伴い，日常生活における物品・サービスの対価の支払決済手段が高度化し多様化する傾向が強まり，最近では，現金授受に代わる電子支払決済が，Fintechと称される実務の重要な一翼を担って展開している。これらの傾向に対応した支払決済法制の再整備は，わが国はもとより世界各国における現代的な主要課題であり，国際的に協調した取組が求められる重要課題でもある。　本稿は，当研究会における共同研究の成果報告の一環として，電子支払決済法制の生成と展開を国際的視野において把握し，重要かつ最新の手掛かりとなる立法資料等を翻訳・紹介する。まず，OECD消費者政策委員会で議論が重ねられ2014年に採択された「OECDモバイル・オンライン決済に関する消費者政策ガイダンス」を翻訳・紹介する。電子支払決済法制整備の国際的なイニシアティブとして，各国での法制整備状況を捕捉する評価軸となる。次に，2015年に改訂された「EU決済サービス指令」（PSD2）を，重要条文を翻訳して紹介する。同改訂指令は，EU各国が同改訂指令にもとづき法制度を具体化する義務を負うことから，欧州各国での法制度整備を予測するための重要な資料であり，さらに，同改訂指令が詳細で優れた手法を提示していることから，欧州各国にとどまらず，わが国を含む多くの国々における立法作業の重要参考資料である。さらに，2016年に成立した大韓民国電子金融取引法改正法について，電子支払決済関連の条文を抜粋して翻訳して紹介する。アジアにおける先進的な取組として，多くの国々の立法作業において注目されるべき試金石である

Applying proteomic methodologies to analyze the effect of hexamethylene bisacetamide (HMBA) on proliferation and differentiation of human gastric carcinoma BGC-823 cells

Human gastric carcinoma BGC-823 cells underwent morphological differentiation and cell cycle arrest in vitro when treated with 5 mM hexamethylene bisacetamide (HMBA) for 48 h. To further understand the mechanism of HMBA-induced differentiation, proteomic methodologies were applied to screen and identify altered proteins involved in the commitment of BGC-823 cells to differentiate. Five distinct altered proteins were acquired by two-dimensional (2-D) PAGE and were consequently identified as ras-related protein rab-35 (Rab-35), splice truncated isoform of transmembrane protease, serine 3 (serine TADG-12), regulator of G-protein signaling 1 (RGS1), ret finger protein-like 1 (RFPL1) and F-actin capping protein alpha-3 subunit (GSG3) by analysis of mass spectrograph. Of the five proteins, serine TADG-12 down-regulated under the detectable level after HMBA treatment, Rab-35, RGS1 and RFPL1 sharply up-regulated within the HMBA-induced BGC-823 cells, and GSG3, appearing in both treated and untreated cells, remarkably increased within BGC-823 cells after HMBA stimulation. Our results implicate that the molecular mechanism of BGC-823 cell differentiation in response to HMBA may involved in complex processes including a signaling network linking vesicle transport, actin cytoskeleton remodeling except for morphology differentiation, cell cycle G1 arrest. (C) 2004 Elsevier Ltd. All rights reserved

Alteration of nuclear matrix-intermediate filament system and differential expression of nuclear matrix proteins during human hepatocarcinoma cell differentiation.

AIM: To investigate the association between the configurational and compositional changes of nuclear matrix and the differentiation of carcinoma cells. METHODS: Cells cultured with or without 5 X 10(-3) mmol/L of hexamethylene bisacetamide (HMBA) on Nickel grids were treated by selective extraction and prepared for whole mount observation under electron microscopy. The samples were examined under transmission electron microscope. Nuclear matrix proteins were selectively extracted and subjected to subcellular proteomics study. The protein expression patterns were analyzed by PDQuest software. Spots of differentially expressed nuclear matrix proteins were excised and subjected to in situ digestion with trypsin. The peptides were analyzed by matrix-assisted laser-desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). Data were submitted for database searching using Mascot tool (www.matrixscience.com). RESULTS: The nuclear matrix (NM) and intermediate filament (IF) in SMMC-7721 hepatocarcinoma cells were found relatively sparse and arranged irregularly. The nuclear lamina was non-uniform, and two kinds of filaments were not tightly connected. After induction for differentiation by HMBA, the NM-IF filaments were concentrated and distributed uniformly. The heterogeneous population of filaments, including highly branched utrathin filaments could also be seen in the regular meshwork. The connection between the two kinds of filaments and the relatively thin, condensed and sharply demarcated lamina composed of intermediate-sized filaments was relatively fastened. Meanwhile, 21 NM proteins changed remarkably during SMMC-7721 cell differentiation. Four proteins, i.e. mutant Pyst1, hypothetical protein, nucleophosmin1, and LBP were downregulated, whereas four other proteins, eIF6, p44 subunit, p-tubulin, and SIN3B were upregulated with the last one, SR2/ASF found only in the differentiated SMMC-7721 cells. CONCLUSION: The induced differentiation of SMMC-7721 cells by HMBA is accompanied by the configurational changes of nuclear matrix-intermediate filament (NM-IF) system and the compositional changes of nuclear matrix protein expression. These changes may be important morphological or functional indications of the cancer cell reversion. (c) 2007 The WJG Press. All rights reserved

Proteomic approach for caudal trauma-induced acute phase proteins reveals that creatine kinase is a key acute phase protein in amphioxus humoral fluid

Elevated creatine kinase (CK) in the circulation was generally regarded to be a passive release from muscle damage. We utilized proteomic methodologies to characterize amphioxus humoral fluid APPs in response to caudal trauma, and found several spots of CK alterations with up-regulation and pi shift. Its. amount and enzyme activity showed a dynamic pattern of APP in humoral fluid companied with a reduction in enzyme activity of muscle, whereas there was no significant difference in CK amount of muscle and the other tissues and in CK enzyme activity of the other tissues between different time points of sample collection following caudal trauma. In addition, CK phosphorylation regulation during injury was not achieved by monoclonal antibodies separately against phosphothreonine, phosphotyrosine, and phosphoserine. These results suggested that the CK elevation of humoral fluid might be from muscle, being an active response to caudal trauma rather than a passive release from muscle damage. Therefore, CK ability in response to caudal trauma should be highly concerned