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Cytokines in adipose-derived mesenchymal stem cells promote the healing of liver disease

World Journal of Stem Cells 2018年第11期10卷 146-159页

作者： Saifun Nahar Yoshiki Nakashima Chika Miyagi-Shiohira Takao Kinjo Zensei Toyoda Naoya Kobayashi issei saitoh Masami Watanabe Hirofumi Noguchi Jiro Fujita Department of Infectious Respiratory and Digestive Medicine Graduate School of Medicine University of the Ryukyus Okinawa 903-0215 Japan Department of Regenerative Medicine Graduate School of Medicine University of the Ryukyus Okinawa 903-0215 Japan Department of Basic Laboratory Sciences School of Health Sciences in the Faculty of Medicine University of the Ryukyus Okinawa 903-0215 Japan Okayama Saidaiji Hospital Okayama 704-8192 Japan Division of Pediatric Dentistry Graduate School of Medical and Dental Science Niigata University Niigata 951-8514 Japan Department of Urology Okayama Univer sity Graduate School of Medicine Dentistry and Pharmaceutical Sciences Okayama 700-8558 Japan

Adipose-derived mesenchymal stem cells （ADSCs） are a treatment cell source for patients with chronic liver injury. ADSCs are characterized by being harvested from the patient’s own subcutaneous adipose tissue, a high cell yield （ i.e. , reduced immune rejection res-ponse）, accumulation at a disease nidus, suppression of excessive immune response, production of various growth factors and cytokines, angiogenic effects, anti-apoptotic effects, and control of immune cells via cell-cell interaction. We previously showed that conditioned medium of ADSCs promoted hepatocyte proliferation and improved the liver function in a mouse model of acute liver failure. Furthermore, as found by many other groups, the administration of ADSCs improved liver tissue fbrosis in a mouse model of liver cirrhosis. A comprehensive protein expression analysis by liquid chromatography with tandem mass spectrometry show-ed that the various cytokines and chemokines produced by ADSCs promote the healing of liver disease. In this review, we examine the ability of expressed protein com-ponents of ADSCs to promote healing in cell therapy for liver disease. Previous studies demonstrated that ADSCs are a treatment cell source for patients with chronic liver injury. This review describes the various cytokines and chemokines produced by ADSCs that promote the healing of liver disease.

关键词： Cell transplantation therapy Cytokine Hepatocytes Liquid chromatography with tandem mass spectrometry Liver cirrhosis Adipose-derived mesenchymal stem cells

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Microbial and enzyme technology: An efficient and convenient method for MiniPrep analysis of recombinant plasmids

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Journal of Biomedical Science and Engineering 2014年第3期7卷 105-107页

作者： Masahiro Sato Emi Inada issei saitoh Yuko Matsumoto Department of Pediatric Dentistry Kagoshima University Graduate School of Medical and Dental Sciences Sakuragaoka Kagoshima Japan Division of Pediatric Dentistry Graduate School of Medical and Dental Science Niigata University Gakkocho-dori Chuo-ku Niigata Japan Section of Gene Expression Regulation Frontier Science Research Center Kagoshima University Sakuragaoka Kagoshima Japan

Minipreparation (MiniPrep) analysis is an essential step for obtaining a recombinant plasmid that carries a DNA insert containing a gene of interest. The most commonly used method for this involves cultivation of transformed Escherichia coli (E. coli) in liquid medium, brief centrifugation for precipitation of bacterial pellets, and subsequent lysis of the pellets. This process is time-consuming and laborious, especially when the sample number is high. Here, we describe a more convenient method for MiniPrep analysis that utilizes solid medium-based cultivation of bacteria.

关键词： Agar Plate E. coli MiniPrep Analysis Gene Engineering Plasmid

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PiggyBac transposon-mediated gene delivery efficiently generates stable transfectants derived from cultured primary human deciduous tooth dental pulp cells(HDDPCs) and HDDPC-derived iPS cells

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International Journal of Oral Science 2015年第3期7卷 144-154页

作者： Emi Inada issei saitoh Satoshi Watanabe Reiji Aoki Hiromi Miura Masato Ohtsuka Tomoya Murakami Tadashi Sawami Youichi Yamasaki Masahiro Sato Department of Pediatric Dentistry Kagoshima University Graduate School of Medical and Dental Sciences Division of Pediatric Dentistry Graduate School of Medical and Dental Science Niigata University Animal Genome Research Unit Division of Animal Science National Institute of Agrobiological Sciences Functional Biomolecules Research Group National Agriculture and Food Research Organization Division of Basic Molecular Science and Molecular Medicine School of Medicine Tokai University Section of Gene Expression Regulation Frontier Science Research Center Kagoshima University

The ability of human deciduous tooth dental pulp cells（HDDPCs） to differentiate into odontoblasts that generate mineralized tissue holds immense potential for therapeutic use in the field of tooth regenerative medicine. Realization of this potential depends on efficient and optimized protocols for the genetic manipulation of HDDPCs. In this study, we demonstrate the use of a Piggy Bac（PB）-based gene transfer system as a method for introducing nonviral transposon DNA into HDDPCs and HDDPC-derived inducible pluripotent stem cells. The transfection efficiency of the PB-based system was significantly greater than previously reported for electroporation-based transfection of plasmid DNA. Using the neomycin resistance gene as a selection marker, HDDPCs were stably transfected at a rate nearly 40-fold higher than that achieved using conventional methods. Using this system, it was also possible to introduce two constructs simultaneously into a single cell. The resulting stable transfectants, expressing td Tomato and enhanced green fluorescent protein, exhibited both red and green fluorescence. The established cell line did not lose the acquired phenotype over three months of culture. Based on our results, we concluded that PB is superior to currently available methods for introducing plasmid DNA into HDDPCs. There may be significant challenges in the direct clinical application of this method for human dental tissue engineering due to safety risks and ethical concerns. However, the high level of transfection achieved with PB may have significant advantages in basic scientific research for dental tissue engineering applications, such as functional studies of genes and proteins. Furthermore, it is a useful tool for the isolation of genetically engineered HDDPC-derived stem cells for studies in tooth regenerative medicine.

关键词： drug selection electroporation genetically modified human deciduous tooth dental pulp cells Piggy Bac

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A simplified protocol for the semi-large scale recovery of plasmids from <i>Escherichia coli</i>grown on agar plates

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Journal of Biomedical Science and Engineering 2012年第7期5卷 406-408页

作者： Masahiro Sato Eri Akasaka issei saitoh Masato Ohtsuka Shingo Nakamura Takayuki Sakurai Satoshi Watanabe Animal Genome Research Unit Division of Animal Science National Institute of Agrobiological Sciences Ibaraki Japan Department of Organ Regeneration Graduate School of Medicine Shinshu University Nagano Japan Department of Pediatric Dentistry Graduate School of Medical and Dental Sciences Kagoshima University Kagoshima Japan Department of Surgery II National Defense Medical College Saitama Japan Division of Basic Molecular Science and Molecular Medicine School of Medicine Tokai University Kanagawa Japan Section of Gene Expression Regulation Frontier Science Research Center Kagoshima University Kagoshima Japan

Semi-large scale liquid cultivation of transformed Escherichia coli (E. coli) in medium (100-200 ml) has been widely used for the acquisition of relatively large amounts of plasmid DNA (50-300 μg). However, this method requires an expensive high-speed centrifugation apparatus to precipitate E. coli before lysis, which is both laborious and time-consuming. Here, we demonstrate a method for agar plate-based cultivation of bacteria that does not employ a high-speed centrifugation apparatus. This procedure proves to be simple and reproducible, yielding an average of 82 μg of plasmid DNA per experiment. It may therefore be valuable for cloning/transfection experiments under limited financial backgrounds.

关键词： Agar Plate DNA Purification E. coli Gene Engineering Plasmid

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A Simplified Protocol for the Semi-Large Scale Recovery of Plasmids from Escherichia coli Grown on Agar Plates

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Journal of Biomedical Science and Engineering 2012年第7期

作者： Masahiro Sato Eri Akasaka issei saitoh Masato Ohtsuka Shingo Nakamura Takayuki Sakurai Satoshi Watanabe

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Improving the performance of conventional base isolation systems by an external variable negative stiffness device under near-fault and long-period ground motions

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Earthquake Engineering and Engineering Vibration 2020年第4期19卷 985-1003页

作者： Sandhya Nepal Masato saitoh Graduate School of Science and Engineering Saitama University255 Shimo-OkuboSakura-kuSaitamaJapan

Recent studies have shown that base-isolated objects with long fundamental natural periods are highly influenced by long-period earthquakes. These long-period waves result in large displacements for isolators, possibly leading to exceedance of the allowable displacement limits. Conventional isolation systems, in general, fail to resist such large displacements. This has prompted the need to modify conventional base isolation systems. The current work focuses on the development of an external device, comprising a unit of negative and positive springs, for improving the performance of conventional base isolation systems. This unit accelerates the change in the stiffness of the isolation system where the stiffness of the positive spring varies linearly in terms of the displacement response of the isolated objects. The target objects of the present study are small structures such as computer servers, sensitive instruments and machinery. Numerical studies show that the increase in the damping of the system and the slope of the linear function is effective in reducing the displacement response. An optimal range of damping values and slope, satisfying the stability condition and the allowable limits of both displacement and acceleration responses when the system is subjected to near-fault and long-period ground motions simultaneously, is proposed.

关键词： base isolation near-fault earthquakes long-period earthquakes negative stiffness variable stiffness

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Model tests and numerical analyses on horizontal impedance functions of inclined single piles embedded in cohesionless soil

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Earthquake Engineering and Engineering Vibration 2013年第1期12卷 143-154页

作者： Chandra Shekhar Goit Masato saitoh Department of Civil and Environmental Engineering Saitama University

Horizontal impedance functions of inclined single piles are measured experimentally for model soil-pile systems with both the effects of local soil nonlinearity and resonant *** practical pile inclinations of 5掳 and 10掳 in addition to a vertical pile embedded in cohesionless soil and subjected to lateral harmonic pile head loadings for a wide range of frequencies are *** obtained with low-to-high amplitude of lateral loadings on model soil-pile systems encased in a laminar shear box show that the local nonlinearities have a profound impact on the horizontal impedance functions of *** impedance functions of inclined piles are found to be smaller than the vertical pile and the values decrease as the angle of pile inclination *** values of horizontal impedance functions are obtained for the ＇positive＇ and ＇negative＇ cycles of harmonic loadings,leading to asymmetric force-displacement relationships for the inclined *** of these experimental results is carried out through three-dimensional nonlinear finite element analyses,and the results from the numerical models are in good agreement with the experimental *** analyses conducted on the numerical models suggest that the consideration of local nonlinearity at the vicinity of the soil-pile interface influence the response of the soil-pile systems.

关键词： inclined single piles harmonic loads horizontal impedance functions local nonlinearity finite element model

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Vertical impedance functions of pile groups under low-to-high loading amplitudes:numerical simulations and experimental validation

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Earthquake Engineering and Engineering Vibration 2023年第3期22卷 647-666页

作者： Usama Zafar Chandra Shekhar Goit Masato saitoh Riku Fukuda School of Science and Engineering Saitama University255 Shimo-OkuboSaitamaJapan

Piles in a group experience additional displacements in soil due to pile-to-pile interactions apart from those resulting from the external *** effect of these interactions determined assuming soil as an elastic and/or viscoelastic material on pile head impedance functions of the pile group is studied by relating the group stiffness to the static stiffness of a single ***,the prevailing elastic solutions may misestimate the resulting pile group response due to the lack of consideration for either soil(material)and/or soil-pile interface *** is well established that soil behaves nonlinearly under moderate-to-high loading amplitudes,and besides,the soil-pile interface nonlinearity can exist even at small loading *** study addresses the effects of these nonlinearities on the vertical impedance functions of a 3×3-pile group using numerical methods by direct analyses and superposition using pile-to-pile interaction *** numerical results are validated using scaled model tests under 1 g *** results highlight the overestimation of pile-to-pile interactions in the pile group when assuming elastic soil *** cases either by direct analyses or superposition approach involving soil and soil-pile interface nonlinearities agree well with the experimental pile group responses under close-to-elastic and nonlinear conditions.

关键词： impedance functions numerical simulations model-scale experiment superposition approach soil-pile interface nonlinearity

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Synchrotron X-ray diffraction techniques for in situ measurement of hydride formation under several gigapascals of hydrogen pressure

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Chinese Science Bulletin 2014年第36期59卷 5290-5301页

作者： Hiroyuki saitoh Akihiko Machida Katsutoshi Aoki Quantum Beam Science Directorate Japan Atomic Energy Agency Sayo-cho Sayo-gun Hyogo 679-5148 Japan Institute for Materials Research Tohoku University Aoba-kuSendai 980-8577 Japan

The high-pressure technique is a fundamental tool for realizing novel phase transitions, chemical reactions, and other exotic phenomena. Hydrogenation is one example of a high-pressure reaction; at high pressures of several gigapascals, hydrogen becomes chemically active and reacts with metals and alloys to form hydrides. This paper covers a high-pressure study of the hydrogenation process and the synthesis of hydrides using a cubic-type multi-anvil apparatus. The experimental details of a hydrogenation cell assembly, high-temperature and highpressure generation, and an in situ observation technique are presented. These experiments are conducted with the aid of in situ synchrotron radiation X-ray diffraction measurements operated in an energy-dispersive mode in the conventional manner for time-resolved measurements and a newly developed angle-dispersive mode for observation of the crystal growth process during formation of metal hydrides. Two successful cases of high-pressure hydrogenation are presented: aluminum hydride, Al H3, and an aluminum-based alloy hydride, Al2 Cu Hx, which are potential candidates for hydrogen storage materials.

关键词： X射线衍射技术金属氢化物原位测量氢气压力同步加速器同步辐射X射线衍射高压技术晶体生长过程

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Influence of Gaps Between Multiple Floating Bodies on Wave Forces

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China Ocean Engineering 2000年第4期15卷 407-422页

作者：缪国平 H. Ishida T. saitoh Professor Department of Civil Engineering Kanazawa University Associate Professor Department of Civil Engineering Kanazawa University

The present study aims to give general hints about hydrodynamic interactions for water wave diffraction on a super large floating structure composed of a large number of box-shaped modules with many small gaps in between. And meanwhile, it also aims to seek for an effective way to take the gap influence into consideration without numerical difficulties existing in conventional methods. An asymptotic matching technique is exploited by virtue of the smallness of gaps. Formal potential solutions are established for the near field around the gap ends and the far field away from gap ends, respectively, and the unknowns in those solutions are uniquely determined by asymptotic matching. The eigen-function expansion method is used for the outer far field and a series of pulsating sources at each gap end is introduced to simulate the gap influence. Strong hydrodynamic interaction is observed and a new resonant phenomenon, the mechanism of which differs absolutely from any known ones, is revealed in the present study. Sharp peak responses for both vertical and horizontal wave-exciting forces on each block are found around some special resonance frequencies, which depend on the draft of the structure and the gap width. The present results are of practical significance to the design of links (connectors) of modules for super large floating structures. And the importance is also closely related to the hydro-elasticity analysis for super large floating structures, in which local loads may be as important as the integrated loads.

关键词： super large floating structure wave interaction resonance gap influence

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