Kyudai Oral Bioscience 2015
―9th International Symposium―OB
K 20152015

February 28th, 2015
Fukuoka Recent Hotel, Fukuoka , JapanPROGRAM & ABSTRACTS

Kyushu University Faculty of Dental Science

〇会期：平成〇会期：平成 27 年 2 月 28 日（土） 9:00～
9:00～13:30〇会場：福岡リーセントホテル〇会場：福岡リーセントホテル 舞鶴の間 A（２階）：下図参照
住所：福岡市東区箱崎 2-5252-1
電話：092
電話：092‐
092‐641‐641‐7741〇主催：九州大学大学院 歯学研究院〇共催：九州大学歯学
共催：九州大学歯学会：九州大学歯学会

連絡先： 中西 博
九州大学大学院 歯学研究院
口腔機能分子科学分野〒812‐
812‐8582 福岡市東区馬出 3-1-1
電話：092
電話：092‐
092‐642‐642‐6413
ファックス：092
ファックス：092‐
092‐642‐642‐6415
Ｅ-mail：mail：nakan@dent.kyushunakan@dent.kyushu-u.ac.jp

1

PROGRAM2

February 28 (Sat) MAIZURU Room A (2nd Floor)
Opening remarks
09:00 - 09:05
09:05 - 09:10

Hiroshi Nakanishi (vice Dean)
Akifumi Akamine (Dean, Faculty of Dental Science, Kyushu University)■Session 1: Keynote Lecture
Chairperson: Fusanori Nishimura (Department of Periodontology)
09:10 - 10:00

Andy Y-T. Teng
Discovery and doubt: a prologue of my quest to research and realism in life
Center for Osteoimmunology & Biotechnology Research (COBR), College of Dental
Medicine, Kaohsiung Medical University and the Affiliated Hospital, Kaohsiung, Taiwan

10:00 - 10:10

Coffee Break■Session 2: Student & PhD Student Session
Chairperson: Yoko Hashimoto (Department of Periodontology)
Fumiko Takayama (Department of Aging Science and Pharmacology)
10:10 - 10:15

Mizuki Abe*
Immunocytochemical detection of glucose-related protein 78 in oral squamouscell carcinoma cell lines
S2-1

Laboratory of Oral Pathology, Faculty of Dental Science, Kyushu University(*MA is a 4th grade undergraduate student of Kyushu University School of Dentistry)
10:17 - 10:22

Katsuhiro Furusho*
The expression of Netrin-1 and its receptor DCC in oral squamous cell
carcinoma
S2-2

Laboratory of Oral Pathology, Faculty of Dental Science, Kyushu University(*KF is a resident of the Section of Dentistry in Kyushu University Hospital)
10:24 - 10:29

Pai-Shien Liao
The clinical efficacy of triclosan-containing toothpaste on the anti-plaque andanti-gingivitis scores: a randomized single-blind clinical study
S2-3

Center for Osteoimmunology & Biotechnology Research (COBR), College of Dental
Medicine, Kaohsiung Medical University and the Affiliated Hospital, Kaohsiung, Taiwan

310:31 - 10:41

Hideki Sugii
Effects of Activin A on the phenotypic properties of human periodontal
ligament cells.
S2-4

Department of Endodontology and Operative Dentistry, Faculty of Dental Science , Kyushu
University
10:43 - 10:53

Kyosuke Toyoda
Grp78 is essential for cell migration induced by amelogenin in a humanperiodontal ligament stem/progenitor cell line.
S2-5

Department of Periodontology, Division of Oral Rehabilitation, Faculty of Dental Science ,Kyushu University
10:55 - 11:05

Yoko Tomita
Strain dependent response of osteocytes in peri-implant bone
S2-6

Section of Implant and Rehabilitative Dentistry, Division of Oral Rehabilitation, Faculty of
Dental Science , Kyushu University
11:07 - 11:17

Miho Ohta
DNA microarray analysis of salivary glands involved in IgG4-related disease.
S2-7

Section of Oral and Maxillofacial Oncology , Kyushu University
11:17 - 11:27

Coffee Break

11:27 - 11:37

Reona Aijima
The role of thermosensitive TRP channel in oral wound healing
S2-8

Department of Molecular Cell Biology and Oral Anatomy, Graduate school of Dental
Science, Kyushu University
11:39 - 11:49

Dan Li
Characterization of a new target Porphyromonas sp. in dental caries prevention
S2-9

Section of Preventive and Public Health Dentistry, Division of Oral Health, Growth andDevelopment, Faculty of Dental Science , Kyushu University
11:51 - 12:01

S2-10 DaGuang Wang

Hetero-oligomerization of C2 domains of phospholipase C-related but
catalytically inactive protein and synaptotagmin-1
Laboratory of Molecular and Cellular Biochemistry, Faculty of Dental Science , Kyushu
University4

12:03 - 12:13

S2-11 Junjun Ni

Cathepsin B deficiency inhibits hypoxic ischemia–induced neuronal damage
through promoting the resolution of neuroinflammation by M2-likepolarization of microglia/macrophages
Department of Aging Science and Pharmacology , Kyushu University
12:15 - 12:25

S2-12 Yi Zhao

The selection of the enteral nutritional support route in neurological intensive
patients
Department of General surgery, Department of Neurosurgery, Peking Union Medical
College Hospital, Beijing, China
12:27 - 12:37

S2-13 Dong-Joon Lee

Gastrin releasing peptide (GRP) expression and its effect tomorpho-differentiation of mouse incisor
Division in Anatomy and Developmental Biology, Department of Oral Biology, Oral
Science Research Center, BK21 PLUS Project, Yonsei University College of Dentistry,Seoul, Korea
12:39 - 12:49

S2-14 Qinghuang Tang

The role of region-distinctive expression of Rac1 in regulating fibronectinarrangement during palatal shelf elevation
Department of Oral Biology, Yonsei University College of Dentistry, Seoul, Korea
12:51 - 12:56

Closing Remarks

5

ABSTRACTS

6

Session 1: Keynote Lecture

7

Discovery and doubt:
a prologue of my quest to research and realism in life
Andy Y-T. Teng
Center for Osteoimmunology & Biotechnology Research (COBR),
College of Dental Medicine, Kaohsiung Medical University
and the Affiliated Hospital, Kaohsiung, Taiwan

Researchers are supposed and expected to create new knowledge and useful information in so-called discovery,as their mission on the jobs, where their outcome is uniquely situated in society and ultimately to the humanhistory. Say: what are and have been the biggest gap(s) in knowledge of life (& our lives)? How about the fact,
the factual, the real, the realism or the truth? What sort of discovery (& doubt in questions) that can, or
ultimately would, be so fundamentally different or real that will lead to the fact …. truth, and thus will stun andchange our futures in life?! Despite, we know better now!? Our living world (the Earth) did not exist at the
time-period of The Big-Bang until about 7.9∼8 billion years ago; so, if we haven't yet identified the beast in thewoods at least we can detect and measure its footprints [2014-Scientific: Dark matter and energy, which weknow nothing about, amount to 95% of all the matter and energy in the observable multiverse that has existed].
So, it was started in 1986 after a DDS training and mandatory military services, where my journey was beefed-up
by Knowing-Nothing for a simple Quest-to-Research in Chicago, along with oral biology/pathology, thenperiodontology, immunology, till osteoimmunology & bone biology, etc., as I did. The factual was that a quarterof a century passed, through which a few things have been quested in the mind-sets via time (why did what I did?
what value(s) defined what I did?): a) what we care for (i.e., desire for, etc.) often qualify, define and constrain
what we are capable of in life (i.e., doctor & patient, treatments & diseases, and the research, too, etc.); b) whatvalues used (in life) to measure the footprints (like the beast in the woods) determine what (& who) we are; c) life
is a means to end in itself, not to achieve the purpose in life; thereafter, d) lacking the inner values (of our mindsets,
in/with our beliefs) often de-ameliorate us and our life itself. Lastly, e) through the ups-and-downs, the hurtles,
and all others I have learned and the not-yet-learned…, this Quest-to-Research was chosen as itself has realized theinner values of myself and the life as I have continued to proceed to further Doubts (more) and Discoveries (less)ahead of me before the next. So, in this presentation, I will share these experiences from the beginning
Quest-to-Research till the next realism to come. After all, the presenter thanks you all and appreciates for theprestige and this opportunity of sharing being offered!!

8Brief biography:
Andy Y-T. Teng is currently a Professor and Director at the Center for Osteo-immunology &Biotechnology Research (COBR), College of Dental Medicine, Kaohsiung Medical University (KMU),
Kaohsiung, Taiwan, and also at the Lab. of Microbial Immunity, Div. of Periodontology, Eastman Institutefor Oral Health (EIOH), University of Rochester, Rochester, USA. He has held a DDS from KMU, Taiwan
and a DMD from University of Rochester, New York, USA, along with MS of Experimental Pathology from
Loyola University of Chicago, USA, and a PhD. of Immunology from University of Toronto, Canada,through which Teng acquired his full-time academic faculty appointments and career for over 24 years inNorth America.
His specialties lie on Periodontology, Pathology and General Dentistry for over 20 years, with much
focused research interests on the immune-regulation of T-&-bone cells’ interactions & cytokine biology,
experimental models of infectious diseases and molecular cell biology, PCR-based diagnostics, and has been
involved in the editorial services of several scientific journals (i.e., JDR, etc.) and for over 40 scientific &professional journals world-wide in the last twenty years.
Prof. Teng has received several international research awards such as the Career-Scientists Award of
Ontario, Canada, International Research Prize of 2003 by the International Congress of Bone and Mineral
Tissue Research, Austria, and 2006-IADR/GSK International -“Innovation Oral Care Research Award”. Also,
Teng has involved in pioneering the field of osteo-immunology and been working with colleagues on themolecular & cellular mechanisms of periodontal diseases, arthritis and osteoporosis as the future therapies forclinical applications.

9Session 2:
Student & PhD Student Session

10

S2-1
Immunocytochemical detection of glucose-related protein 78 in oral squamous cell
carcinoma cell lines
Mizuki Abe*, Kengo Nagata, Akiko Jinno, Tamotsu Kiyoshima
Laboratory of Oral Pathology, Faculty of Dental Science, Kyushu University(*MA is a 4th grade undergraduate student of Kyushu University School of Dentistry)S2-2
The expression of Netrin-1 and its receptor DCC in oral squamous cell carcinoma
Katsuhiro Furusho*, Hiroko Wada, Yurie Mikami, Tamotsu Kiyoshima
Laboratory of Oral Pathology, Faculty of Dental Science, Kyushu University(*KF is a resident of the Section of Dentistry in Kyushu University Hospital)

11

S2-3
The clinical efficacy of triclosan-containing toothpaste on the anti-plaque andanti-gingivitis scores: a randomized single-blind clinical study
Pai- Shien Liao*1 & Andy Y-T. Teng1
1.

Center for Osteoimmunology & Biotechnology Research (COBR), College of Dental Medicine, Kaohsiung
Medical University and the Affiliated Hospital, Kaohsiung, Taiwan

Background: Triclosan has been shown to mediate certain anti-microbial effects and modulation of theimmune responses. To assess its clinical efficacy, we rationalized and designed a single-blinded randomization
clinical study based on its effects. Methods: Sixty male and female subjects with gingivitis and a minimal of 17
teeth present, who have met the inclusion and exclusion criteria, are recruited and consented as well, based on
IRB protocols with approval. Subjects are examined at baseline for the oral indexes including Loe & Silness(GI) and a plaque index (Turesky modified Quigley & Hein (PI). Subjects are informed of possible adverse
reactions that they could experience, such as oral irritations. Then, all subjects are blinded and randomly
subjected to receive a Tricosan-containing toothpaste or another tooth-paste containing separate control
ingredients without Triclosan. All subjects are instructed with standardized protocol to brush their teeth twice
daily for 6 weeks. At the end of 6 weeks, an exit-examination is carried out again for GI & PI, through which a
two factor ANOVA will be used to determine if the differences between the above treatments are significant andthe differences are considered significant if a 95% confidence level is achieved. Summary: The present study is
ongoing without the results yet; however, it is anticipated that certain tooth-paste ingredients like triclosan, in its
technology under test, is to manifest certain anti-microbial effects in human subjects with gingivitis for at least ashort-term 6-week effect. Note: The study has been funded by Center for Osteoimmunology & Biotechnology
Res (COBR), College of Dental Medicine, Kaohsiung Medical University, and a pilot fund from theColgate-Palmolive Co., Global & Taiwan divisions (S102023).

12

S2-4
Effects of Activin A on the phenotypic properties of human periodontal ligament cells
Hideki Sugii1, Hidefumi Maeda2, Atsushi Tomokiyo2, Naohisa Wada2, Satoshi
Monnouchi1, Daigaku Hasegawa1, Sayuri Hamano1, Asuka Yuda1, Shinichiro Yoshida1,
Suguru Serita1, Hiroyuki Mizumachi1,Hiromi Mitarai1, Akifumi Akamine1,2.
1Department of Endodontology and Operative Dentistry, Faculty of Dental Science, Kyushu University, and2
Department of Endodontology, Kyushu University Hospital

Activin A (ACTA) that is a member of the TGF-β superfamily and a dimer of inhibinβa contributes to tissue
healing. It was reported that ACTA exerts an inhibitory effect on osteoblastic differentiation and mineralization.
However, the expression and function of ACTA in human periodontal ligament cells (HPDLCs) have not beenreported. Thus, we aimed to analyze the biological effects of ACTA on HPDLCs.
HPDLCs were isolated from healthy third molars. HPDLCs were cultured with or without ACTA in thepresence or absence of CaCl2 that was used for inducing osteoblastic differentiation. The cells were thensubjected to Alizarin red S staining and von Kossa staining. Gene expressions were evaluated by quantitative
RT-PCR, and protein expressions were investigated by immunofluorescence (IF) and immunochemical (IC)analyses. Chemotaxis, migration and proliferation of HPDLCs treated by ACTA were also examined using
modified Boyden Chamber assay, scratch wound healing assay and WST-1 proliferation assay, respectively.
IC analysis with an anti-ACTA antibody revealed positive staining in HPDLCs and rat PDL tissue. When rat
PDL tissue was experimentally damaged, IF results showed that positive reactions against both anti-ACTA andanti-IL-1β antibodies were up-regulated around the damaged sites where co-localized detection was seen.
ACTA promoted chemotaxis, migration and proliferation of HPDLCs. Quantitative RT-PCR data showed that
this protein induced the expression of genes related to fibroblastic differentiation, but inhibited theosteoblast-related gene expression in HPDLCs. ACTA furthermore inhibited mineralization of CaCl2-treated
HPDLCs.
Our results suggested that ACTA was expressed in the entire PDL tissue at low levels while its expression was
promoted during the early phase of damaged PDL tissue healing. In addition, ACTA could promote
proliferation, migration, chemotaxis and fibroblastic differentiation of PDL cells whereas inhibited theirosteoblastic differentiation. This protein might have the role to prevent ectopic PDL tissue ossification byblocking osteoblastic differentiation of PDL cells. Therefore, ACTA could be applied as a therapeutic factor that
promotes healing and regenerating of PDL tissue damaged by disease, trauma or surgical reconstruction.

13

S2-5
Grp78 is essential for cell migration induced by amelogenin in a human periodontal
ligament stem/progenitor cell line.
Kyosuke Toyoda, Takao Fukuda, Terukaze Sanui, Kensuke Yamamichi, Ryo Atomura,
Urara Tanaka, and Fusanori Nishimura.
Department of Periodontology, Division of Oral Rehabilitation, Faculty of Dental Science, Kyushu
University.
Objective
The major component of enamel matrix proteins, amelogenin is a potential bioactive molecule for periodontal
regeneration, although the downstream target molecules and/or signaling are still unknown. Recently, we
performed proteomics analysis to elucidate amelogenin-interacting networks. We identified Grp78 as a newamelogenin binding protein, and the biological interaction of amelogenin with this molecule enhanced cellproliferation in osteoblastic cells (Fukuda et al, 2013). In this study, since specific migration and proliferation of
periodontal ligament cells (PDL) play key role in successful periodontal regeneration, we evaluated thebiological interaction between amelogenin and Grp78, and its effect on cellular responses in periodontal
ligament (PDL) cells.
MethodAn established multipotent clonal human PDL stem/progenitor cell line (PDLSCs), 1-17, was used. GST pull
down assay and confocal co-localization were performed to validate the binding of recombinant amelogenin(rM180:10µg/mL) with endogenous Grp78. Grp78 was overexpressed or knock-downed in PDLSCs for further
characterization of these molecular interactions to observe gene expression profiles (microarray, pathway, andheatmap analysis), cell motility (wound healing and boyden chamber assay), cell viability (WST-8, Ki-67
positive staining), Rho signal transductions (Rho family activation), and morphological changes (confocal
microscopy).
ResultInternalization of rM180 via binding to the cell surface Grp78 was observed in PDLSCs. Microarray analysis
demonstrated that rM180 activates TGF-β pathway, and Grp78 altered the expression of several cell migration
related genes. Overexpression of Grp78 enhanced rM180-induced cell migration and adhesion without
affecting cell proliferation, while silencing of Grp78 diminished these activities. Binding of rM180 with Grp78
promoted lamellipodia formation, and the simultaneous activation of Rac1 was observed.
Discussion
The results suggested that amelogenin binding with Grp78 is involved in the regulation of cell migration,
moreover, the Rac1 activation and lamellipodia formation are critical steps for amelogenin-induced cell
migration.

14

S2-6
Strain dependent response of osteocytes in peri-implant bone
Yoko Tomita1, Yasuko Moriyama1, Yasunori Ayukawa1, Kosaku Kurata2, Takanobu
Fukunaga2, Hiroshi Takamatsu2, Kiyoshi Koyano1
1Section of Implant and Rehabilitative Dentistry, Division of Oral Rehabilitation, Faculty of Dental Science,
Kyushu University
2
Department of Mechanical Engineering, Faculty of Engineering, Kyushu University

Osteocytes are thought to be mechanosensors and responsible for orchestrating bone remodeling process.
However, little is known about how osteocytes react to mechanical stress received from dental implant. The
purpose of this study was to investigate the effects of strain-induced biochemical response of osteocytes in vitro.
We established two types of loading apparatuses, in which MLO-Y4 osteocyte cell line wasthree-dimensionally cultured, subjected to the application of cyclic mechanical stress in physiological strain for
24 hours. One could simultaneously apply five kinds of (from 0.095% to 1.22%) stretching strains and the other
could reciprocate titanium plate, which imitated dental implant screw shape, in the cell-embedded gel. Cyclic
physiological strain applied by both devices induced the cell death of MLO-Y4. Real-time RT-PCR analyses
showed that the mechanical strain increased RANKL/OPG ratio and suppressed sclerostin expression in amagnitude-dependent manner. Connexin43 expression increased in the middle strain magnitude. The
conditioned medium obtained from low magnitude cyclic stretch culture increased ALP activity of bonemarrow cell culture.
These studies suggest that osteocytes may respond to strain from implant and coordinate peri-implant bone
remodeling.

15

S2-7
DNA microarray analysis of salivary glands involved in IgG4-related disease.
Miho Ohta, Masafumi Moriyama, Akihiko Tanaka, Takashi Maehara, Sachiko
Furukawa, Shintaro Kawano, Jun-Nosuke Hayashida, and Seiji Nakamura
Section of Oral and Maxillofacial Oncology, Kyushu University
Objectives. IgG4-related disease (IgG4-RD) is a systemic disease characterized by infiltration of IgG4-positive
plasma cells with severe fibrosis in multiple target organs. Our previous reports demonstrated that helper T type2 (Th2) cells are involved in IgG4 production of IgG4-RD. Moreover, Watanabe et al. reported that abnormal
innate immune responses might enhance Th2 immune responses and the immunopathogenesis of IgG4-RD. In
this study, we thus addressed to identify the disease-associated genes, especially innate immune molecules,
using DNA microarray.
Methods. Gene expression was analyzed by DNA microarray in submandibular gland (SMG) from patientswith IgG4-RD (n=3) and healthy controls (n=3). Differentially expressed genes (DEGs) between the twogroups were identified, and gene-annotation enrichment analysis was performed by using Gene Ontology (GO)annotation. Validation of the results was performed by quantitative PCR and immunohistochemistry using
salivary glands from IgG4-RD (n=7), Sjögren’s syndrome (n=10), and healthy controls (n=10).
Results. Gene expression patterns in IgG4-RD and healthy controls were quite different with each other inhierarchical clustering as well as principal component analysis. In IgG4-RD, 450 up-regulated genes and 732
down-regulated genes were identified as DEGs (adjust p-value <0.01). GO term analysis indicated that the
up-regulated genes of DEGs in IgG4-RD encoded proteins that function in T/B cell activation, and chemotaxis.
PCR validated significantly higher expression of “macrophage receptor with collagenous structure (MARCO)”
in IgG4-RD than that in controls (p<0.001). MARCO belongs to Class A scavenger receptor and expressed on“alternatively activated (M2) macrophages” which is activated by Th2 cytokines can induce wound healing andfibrosis. Immunohistochemical analysis confirmed that the number and the frequency of M2 macrophages in
IgG4-RD were significantly higher than those in the other groups.
Conclusion. MARCO was identified as a disease-associated gene and might be critically important inelucidating the relationship between innate immunity and the pathogenesis of IgG4-RD.

16

S2-8
The role of thermosensitive TRP channel in oral wound healing
Reona Aijima1,2,3, Bing Wang1, Tomoka Takao1, Hiroshi Mihara4, Makiko Kashio4,
Yasuyoshi Ohsaki1, Jing-Qi Zhang1, Sadahiko Masuko2, Makoto Tominaga4, Mizuho A.
Kido1

1

Department of Molecular Cell Biology and Oral Anatomy,
Graduate school of Dental Science, Kyushu University
2
Department of Oral and Maxillofacial Surgery, Faculty of Medicine, Saga University
3
Division of Histology and Neuroanatomy, Department of Anatomy and Physiology,
Faculty of Medicine, Saga University
4
Division of Cell Signaling, Okazaki Institute for Integrative Bioscience (National Institute for Physiological
Sciences), National Institutes of Natural Sciences
Oral cavity, the entrance to the alimentary tract, experiences drastic changes of temperature, chemical ormechanical stimuli, compared with other tissues. Because of this, oral mucosa is susceptible to injury, but it is
known to provide faster wound healing than the skin and less scar formation. However, the molecular bases that
regulate this wound healing are still unclear.
Recently, there has been accumulating evidence that thermosensitive transient receptor potential (TRP)
channels contribute to not only environmental sensors but also physiological functions. We found TRP vanilloid
3 (TRPV3), a thermosensitive Ca2+-permeable channel activated by warm temperatures (above 33°C), wasfunctionally expressed in oral epithelia. Using a molar tooth extraction model, TRPV3-deficient (TRPV3KO)
mice showed delayed wound closures compared with those in wild-type (WT) mice. Furthermore, prominent
up-regulation of TRPV3 mRNA was found in the wounded tissues at an early stage of wounding. TRPV3
agonist stimulation or temperature above 33°C to primary cultured oral epithelial cells enhanced the number ofproliferating cells. In addition, the activation of TRPV3 increased phosphorylation of epidermal growth factor
receptor (EGFR) in the cells from WT, which was not found in the cells from TRPV3KO.
These results suggest TRPV3 in oral epithelia promotes the proliferation of oral epithelial cells and contributes
to rapid wound repair via EGFR phosphorylation. Our findings suggest that TRPV3 activation could be apotential therapeutic target for wound healing in skin and oral mucosa.
Aijima R, Wang B, Takao T, Mihara H, Kashio M, Ohsaki Y, Zhang JQ, Mizuno A, Suzuki M, Yamashita Y,
Masuko S, Goto M, Tominaga M, Kido MA. FASEB J. 2015 29(1):182-92.

17

S2-9
Characterization of a new target Porphyromonas sp. in dental caries prevention
Dan Li, Yukie Shibata, Toru Takeshita, Yoshihisa Yamashita
Section of Preventive and Public Health Dentistry, Division of Oral Health, Growth and Development,
Faculty of Dental Science, Kyushu University, Fukuoka, Japan
Dental caries forms by cariogenic virulent bacteria through complex interactions among oral microbiota.
Although the causative agents of dental caries have been extensively studied, little attention was given tocomplex and forming process of healthy plaque. Our previous study with pyrosequencing analysis indicatedthat the prevalence ratio of an oral taxonomic unit (OTU) assigned to genus Porphyromonas showed a higher
tendency in plaque of caries-free subjects than in that of caries-experienced ones during a 7-day in vivo
experiment. The OTU presented a colonization mode similar to a middle colonizer, Fusobacterium species andwas significantly more predominant in saliva of caries-free subjects than in that of caries-experienced ones.
In the present study, in order to further confirm the results of the pyrosequencing analysis, quantitative realtime PCR and clone library were performed. The results of quantitative real time PCR of saliva and day-7
plaque were consistent with that of the pyrosequencing analysis. Clone library analysis revealed that an oralclone CW034 resembler which belong to Porphyromonas sp. HOT-279 was identified as the predominant
species in caries free subjects of both in saliva and day-7 plaque. We isolated a bacterial strain resembling anoral clone CW034 from human saliva and characterized its biological property. It was weakly saccharolytic anddid not form pigmented colonies, being different from Porphyromonas gingivalis, a periodontal pathogen, in
general. Based on characterized its property, we proposed to name it Porphyromonas pasteri KUFDS01.
Fusobacterium species are considered important “bridge” organisms that coaggregate initial, early, and latecolonizers, and contribute to an increase in microbial diversity in plaque development. We compared the abilityof coaggregation of Fusobacterium nucleatum ATCC 10953 and P. pasteri KUFDS01, and P. pasteri
KUFDS01 and F. nucleatum ATCC 10953 differed in coaggregation ability.
These results suggested P. pasteri KUFDS01 is a new target to elucidate the mechanism of plaque biofilm
formation. To investigate ‘healthy plaque’ is a novel viewpoint and meaningful for examining the progress ofdental caries.

18

S2-10
Hetero-oligomerization of C2 domains of phospholipase C-related but catalyticallyinactive protein and synaptotagmin-1
DaGuang Wang1, Hiroshi Takeuchi2, Jing Gao1, Zhao Zhang1,3, Masato Hirata1
1

Laboratory of Molecular and Cellular Biochemistry, Faculty of Dental Science, Kyushu University,
2
Division of Applied Pharmacology, Kyushu Dental University,
3
Stomatological Hospital of Hebei Medical University,

The C2 domain is a protein module often found in molecules that regulate exocytosis. C2 domains mediate
interactions between the parental molecule and Ca2+, phospholipids, and proteins. Although various molecules
have been shown to interact with several C2 domains, no interactions between the C2 domains from different
molecules have yet been reported. In the present study, we identified direct interactions between the C2 domain of
PRIP (phospholipase C-related but catalytically inactive protein) and the C2 domains of other molecules. Among
the C2 domains examined, those of synaptotagmin-1 (Syt1-C2A and Syt1-C2B) and phospholipase C δ-1 boundto the C2 domain of PRIP. We investigated the interactions between the C2 domain of PRIP (PRIP-C2) with
Syt1-C2A and Syt1-C2B, and the mode of binding of each was Ca2+-dependent and -independent, respectively.
We further demonstrated that the Ca2+ dependence of the interaction between PRIP-C2 and Syt1-C2A wasattributed to Ca2+ binding with Syt1-C2A, but not PRIP-C2, using a series of mutants prepared from both C2
domains. We previously reported that the interaction between PRIP-C2 and the membrane fusion machinerysuggested a critical role for PRIP in exocytosis; therefore, the results of the present study further support theimportance of PRIP-C2 in the inhibitory function of PRIP in regulating exocytosis.

19S2-11Cathepsin B deficiency inhibits hypoxic ischemia–induced neuronal damage throughpromoting the resolution of neuroinflammation by M2-like polarization of
microglia/macrophages
Junjun Ni, Zhou Wu, Hiroshi Nakanishi
Department of Aging Science and Pharmacology, Kyushu University, Fukuoka, Japan
It is widely accepted that microglia/macrophages acquire distinct functional phenotypes: classically (M1) oralternatively activated (M2) microglia/macrophages. M1- and M2-like microglia/macrophages respectively
dominate in the chronic and acute states of neuroinflammation. In spite of clinical importance, little is known about
the mechanisms underlying phenotypic shift of microglia/macrophages. Here, we provide the first evidence thatcathepsin B (CatB), a typical cysteine lysosomal protease, can work as a phenotype switch of
microglia/macrophages along the M1-M2 phenotypic continuum. Neonatal wild-type mice subjected tohypoxic/ischemia (HI) injury showed an extensive brain injury. CatB was significantly increased exclusively in
microglia/macrophages after HI injury. It was also noted that microglia/macrophages showed early and persistent
M1-like polarization followed by transient M2-like polarization after HI injury. In neonatal CatB-/- mice, however,HI-induced brain injury was significantly reduced. Furthermore, CatB-deficient microglia/macrophages showed
only the early and transient M2-like, but not M1-like, polarization. When conditioned medium from M1-polarized
microglia (M1-CM) was prepared in the presence of CA-074Me, a specific inhibitor of CatB, the neuronal deathof primary cultured hippocampal neurons induced by M1-CM was significantly declined. Furthermore,
CA-074Me prevented activation of nuclear factor-κB (NF-κB) through inhibiting autophagic degradation of IκBαin microglia following oxygen glucose deprivation. These observations suggest that CatB is a potential phenotype
switch for promoting M1-like polarization of microglia/macrophages through prolonged activation of NF-κB, thus
inhibition of CatB activity, either through genetic deletion a pharmacological inhibitor, ameliorates HI-induced
brain injury via promoting M2-like polarization of microglia/macrophages.
Keywords: microglia/macrophage, phenotype switch, hypoxic-ischemia

20

S2-12The selection of the enteral nutritional support route in neurological intensive patientsYi Zhao1,2, Jianchun Yu1, Renzhi Wang3, Zhiqiang Ma4
1

Dept. of General surgery, Peking Union Medical College Hospital2
Dept. of Neurosurgery, Peking Union Medical College Hospital3
Dept. of Neurosurgery, No.1 Hospital of Jilin University
4
Dept. of Neurology, No.1 Hospital of Jilin University

Background:
Neurological patients suffered from swallow disorders in acute or chronic neurological diseases such as skull
fracture, stroke, and motor neuron disease and so on, have more nutritional risks, complications and mortality.
Early and adequate nutritional support is thought to improve neurological outcome and decrease morbidity. The
selection of the enteral tube feeding route by oral or nasal gastric tube (NGT) or percutaneous endoscopic
gastrostomy (PEG) for long-term feeding issues is controversy.
Objective:
1. To compare the influence of normal format meal and non-normal format meal to the nutritional condition,complication and prognosis of patients with a nasogastric tube suffering from neurological diseases.
2. Retrospective analysis of the influence of PEG treatment to gender ratio, distribution of etiology, turnover,reason of death, and complications of the patients.
3. Compare incidence rate of nutrition risk, nutrition index and complications between patients whose GCS
was less than 8 and those whose GCS not less than 8.
Methods:
1. Admitted patients who have swallowing disorder and with nutritional risk were treated with NGT therapy,
being divided by two groups (group A and Group B, each group contains 9 patients). Patients in group A weretreated with normal format meal which was given 1125kcal/d, with 48g protein per day in the first 3d, and given
1500kcal/d, with 64g protein per day after 3d, while those in Group B were treated with non-normal format
meal which was given less than 1125kcal/d, with no more than 32g protein per day in the first 3d, and given lessthan or more than 1500kcal/d, with more than or less than 64g protein per day after 3d. Nutrition index, immune
index, and complications of three periods (before intubation, 1w after intubation and 2w after intubation) wereobserved.
2. Retrospective analysis of 72 cases of PEG treatment of neurological patients was performed, in which
patients’ gender ratio, distribution of etiology, turnover, reason of death, complications were analyzed.
3. Divide the 32 admitted patients into 2 groups, one is whose GCS was less than 8, and the other is whose
GCS was not less than 8. Compare incidence rate of nutrition risk as soon as they came to hospital, and nutritionindex and complications when they came to hospital and a week later between patients whose GCS was lessthan 8 and those whose GCS not less than 8.

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Result:
1.As soon as they came to hospital, the serum Creatinine of Group A is 72.0(55.0-82.5), of Group B is50.3(32.5-63.7), with p=0.027(p<0.05); 1w later, the CHOL of Group A is 3.05(2.88-3.90), of Group B is
3.98(3.83-4.48), which is significant with p=0.031(p<0.05). 2w later, the BUN of Group A is 6.89(5.32-7.34), ofGroup B is 4.33(3.11-5.51), with p=0.009(<0.05); the serum Creatinine of Group A is 56.0(42.5-66.0), ofGroup B is 37.5(30.4-51.0), p=0.047(<0.05). However, the incidence of other immune index and complications
does not have statistical significance.
2. The average age of patients in the research is 73.1±15.5 yrs old, male to female 1.77:1. Those above 80 werein the largest number 29(40.3%) were mainly suffering from cerebral thromboembolism 35(48.6%) and Motorneuron disease 21(29.2%). The total number of patients are 72, with 63 of whom ( 87.5%) improved, 9 ofwhom died ( 12.5%) while most of death cases of 7 (77.8%) were due to pneumonia. 65 patients were foundwith complications which contains 34 aspirated pneumonia (47.2%) and 51 swallowing difficulty (70.8%).
3. Among the admitted 32 patients, there are 3 patients of GCS less than 8 who had the complications (33.3%),
6 patients without any complication(66.7%), none of patients whose GCS was not less than 8 points had any
complications, p<0.05. At the time of admission, great significance was found among the patients from twogroups, p=0.032 (<0.05). The calf girth of patients of GCS<8 is (36.0±1.1) cm , and is (34.8±1.3) cm of GCS≥8,
with p=0.032(<0.05).
Conclusion: Treating patients suffering from neurological diseases with nutritional risk with normal-format
meal can significantly ameliorate their index of BUN and serum Creatinine, however cannot lower theincidence of complications, there are no significant difference among other nutritional indicators. PEG has moremeanings for old age patients, pneumonia is the chief cause for bad prognosis. Patients whose GCS is less than
8 have more probability in nutrition risk and complications than those whose GCS is not less than 8.

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S2-13
Gastrin releasing peptide (GRP) expression and its effect to morpho-differentiation ofmouse incisor
Dong-Joon Lee, Han-Sung Jung
Division in Anatomy and Developmental Biology, Department of Oral Biology, Oral Science ResearchCenter, BK21 PLUS Project, Yonsei University College of Dentistry, 50 Yonsei-Ro, Seodaemun-Gu, Seoul,
120-752, Korea.
The gastrin-releasing peptide (GRP), and its receptor (GRPR) are widely distributed in mammalian peripheral
tissues and in the central nervous system. Effects of GRP and its receptor on the production and release ofcytokines are described both in animal models and humans with inflammatory diseases. Moreover, GRP is amajor growth factor in many types of human cancers. The GRPR is overexpressed in different malignancies andits activation stimulates tumor growth. And GRP oncogenic signaling pathway was turned out relatively
detailed. Thus, GRPR antagonists have been developed as candidate anticancer agents and shown to display
anti-proliferative activity in experimental cancer models. Recent studies of GRP and GRPR are concentrated on
gastrointestinal tract, pulmonary region and central nervous system. However, the function of GRP and GRPR
is still poorly revealed in the oro-facial region.
The GRPR is G-protein coupled receptor and 7-transmembrane glycosylated receptor that activates thephospholipase C signaling pathway. And the GRP regulates mobilization of calcium from intracelluar stores.
We suggest that GRP can be one of the factors which affect regulation of calcification during tooth development
based on these previous studies.
This study was supported by BK21 PLUS Project, College of Dentistry, Yonsei University23

S2-14
The role of region-distinctive expression of Rac1 in regulating fibronectin arrangement
during palatal shelf elevation
Qinghuang Tang, Liwen Li, Chengri Jin, Jong-Min Lee, Han-Sung Jung
Department of Oral Biology, Yonsei University College of Dentistry, Seoul, Korea
Palatal shelf elevation is a crucial process in palate development, contributed by various factors. Disturbances inany factor during this process result in cleft palate. Prior to palatal shelf elevation, started from E12.5, higher
Rac1 expression level in bend region and increased mesenchymal cell density in the bend and groove regions of
mid-palatal shelf were observed. The high cell densities in the bend and groove regions correlated with aring-like arrangement of fibronectin (FN), whereas low cell density in middle region correlated with a fibrillar
FN arrangement. Rac1 overexpression altered the fibrillar FN arrangement in the middle region to the ring-likearrangement. This alteration was sufficient to induce the failure of palatal shelf elevation and, ultimately leading
to cleft palate. Furthermore, inhibition of FN delayed palatal shelf elevation. Our results indicate that theregional expression of Rac1 played an impressive role in palatal shelf elevation, by regulating the arrangementof FN within the palatal shelf.
Key words: Rac1, Fibronectin, Palatal shelf, Elevation, Cleft palate

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K 20152015