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Distribution and Ecology of Marsh Clam in Gyeongsangbuk-do II. Reproductive Cycle and Larval Development of the Corbicula japonica (경상북도 재첩자원 분포 및 생태 조사 II. 일본재첩 Corbicula japonica의 생식주기 및 유생발생)

  • 변경숙;정의영
    • The Korean Journal of Malacology
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    • v.17 no.1
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    • pp.45-55
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    • 2001
  • Gametogenes, reproductive cycle, first sexual maturity(biological minimum size), sex ratio and larval development of the marsh clam Corbicula japonica were investigated monthly by histological observations. Samples were collected in brackish water of Gokgang stream, Kyungsangbuk-Do, Korea, from August 1997 to July 1998. Sexuality of Corbicula japonica is dioecious and the species are an oviparous clam. The gonads are irregularly arranged from the sub-region of mid-intestinal gland in visceral cavity to reticular connective tissue of foot. The ovary is composed of a number of ovarian sac which are branched arborescent. Oogonia actively proliferate along the germinal epithelium of ovarian sac, in which young oocytes are growing. The testis is composed of a number of testicular tubules, and the epithelium of the tubule has function of germinal epithelium, along which spermatogonia actively proliferate. A great number of undifferentiated mesenchymal tissue and eosinophilic granular cells are abundantly distributed between developing oocytes and spermatocytes in the early developmental stages. With the further development of the ovary and testis these tissue and cells gradually disappear. Then the undifferentiated mesenchymal tissue and eosinophilic granular cells are considered to be related to the growing of the oocytes and spermatocytes. The spawning period is from July to September, and the main spawning occur between July and August when seawater temperatures reach above 22$^{\circ}C$. The reproductive cycle of this species can be divided into five successive stages; early active (February to April), late active (May to July), ripe (June to September), partially spawned (July to September), degenerative (September to October) and resting stage (October to February). Percentages of first sexual maturity of female and male clams ranging in length from 10 mm to 12 mm are over 50% and 100% for clams over 16.0 mm in shell length. Fertilized eggs or Corbicula japonica were 80-90 ${\mu}{\textrm}{m}$ in diameter. In the early embryonic development of C. japonica, the appearance of polar body, trochophore and D-shaped veliger were observed around 40 min., 27 hours and 4 days after spawning, respectively, at a water temperature of 26.5-28.$0^{\circ}C$. The size of larvae of early umbo stage was about 185-210 ${\mu}{\textrm}{m}$ in shell length, 160-180 ${\mu}{\textrm}{m}$ in shell height around 7 days after fertilization. The correlation of relative growth between the culture day (D) and shell length (SL) was expressed by the following simple formula from D-shaped veliger to metamorphosing stage; SL = 13.300D + 209.36($r^2$= 0.9078).

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Effects of Glycyrrhiza inflata Batal Extracts on Adipocyte and Osteoblast Differentiation (감초추출물의 지방세포와 조골세포에 대한 분화효과)

  • Seo, Cho-Rong;Byun, Jong Seon;An, Jae Jin;Lee, JaeHwan;Hong, Joung-Woo;Jang, Sang Ho;Park, Kye Won
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.7
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    • pp.1015-1021
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    • 2013
  • Glycyrrhiza inflata Batal, an important species of licorice, is one of the most widely used medicinal plants for over 4000 years. Glycyrrhiza plant species has been well known for its various therapeutic activities such as anti-inflammatory, anti-allergic, and anti-ulcer. The purpose of this study was to determine the effects of Glycyrrhiza inflata Batal ethanol extracts (GBE) on adipocyte and osteoblast differentiation. Mesenchymal C3H10T1/2 cells were treated with sub-cytotoxic doses of GBE, and its effects on adipocyte differentiation were assessed. We found that GBE dose-dependently increased lipid accumulation and also induced the expression of adipocyte markers, such as $PPAR{\gamma}$ and its target genes, aP2, and adiponectin, in C3H10T1/2 cells. Consistently, similar effects of GBE on lipid accumulation were also observed in preadipocyte 3T3-L1 cells that further supports the pro-adipogenic activities of GBE. We also investigated the effects of GBE on osteoblast differentiation of mesenchymal C3H10T1/2 cells. As a results, we found that GBE increased the activity of alkaline phosphatase in a dose-dependent manner and also promoted the expression of osteoblast markers, such as ALP and RUNX2, during osteoblast differentiation of C3H10T1/2 cells. Similar pro-osteogenic effects of GBE were also observed in preosteoblast MC3T3-E1 cells. Finally, our data show that a major bioactive compound found in Glycyrrhiza inflata Batal, licochalcone A (LA) but not glycyrrhizic acid (GA), can mediate the pro-adipogenic and pro-osteogenic effects of GBE. Taken together, this study provides data to show the possibility of GBE and its bioactive component LA as putative strategies for type 2 diabetes and bone diseases.

Gametogenesis and Reproductive Cycle of the Cockle, Fulvia mutica (Reeve) (새조개, Fulvia mutica (Reeve)의 생식세포형성과정 및 생식주기)

  • CHANG Young Jin;LEE Taek Yuil
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.15 no.3
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    • pp.241-250
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    • 1982
  • The structure of gonads, gametogenesis and reproductive cycle of the cockle, Fulvia mutice, were studied mainly by histological observation. The materials were monthly sampled in the southern area of Yeosu from October 1980 to September 1981. F. mutica was monoecious. The gonads were situated between the liver tissues and the outer fibronuscular layers compacted by the connective tissue fibers and muscle fibers beneath the outermost layer of simple cuboidal epithelium. The gonad was composed of a number of the ovarian sacs and the testicular tubules which form the tubular structure. Testicular tubules in the mature stage sometimes contained 'testis-ova' The undifferentiated mesenchymal tissues and the eosinophilic cells were abundantly distributed on the germinal epithelium in the early development stage. With the further development of the ovary and testis, these tissues and cells gradually disapprared. The undifferentiated mesenchymal tissues and the eosinophilic cells are related to the growing of the oocytes and spermatocytes . Early multiplicating oogonium was about $10{\mu}m$ in diameter. As the oocytes grow to $27-34\times50-58{\mu}m$ by increasing cytoplasm, the oocytes connected to the basement membrane by their egg-stalks. The ripe eggs were about $60{\mu}m$ in diameter and they were surrounded by gelatinous membrane. Most male germ cells in mature stage were transformed into the spermatozoa and they formed the sperm bundles. After spawning, undischarged ripe eggs and spermatozoa remained in the ovarian sac and the testicular tubule respectively for some time, then they finally degenerated. Especially the early spent ovarian sacs in May did not contract significantly and then they took part in the secondary maturation within two or three months during the summer season. The monthly changes of the fatness well agreed with the reproductive cycle. The reproductive cycle of F. mutica could be classified into six successive stages : multiplicative, growing, mature, spent, degenerative and recovery stage. It seems that the spawning season is closely rotated to the water temperature, and the spawning occurs from May to October at about $20^{\circ}C$ in water temperature. The peak spawning seasons appeared twice a year between June and July and in September. Acknowledgement The authors wish to express their gratitude to Dr. Kim, In Bae, Dr. Chun, Seh Kyu and Dr. Yoo, Sung Kyoo of National Fisheries University of Busan and Mr. Min, Byoung Seo of National fisheries Research and Development Agency for their critical reading of the manu script.

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Retrospective Investigation of Canine Skin and Mammary Tumors in Korea (한국에서 개 유선종양과 피부종양의 발생현황)

  • Kim, Yeong-Hun;Ahn, Na-Kyoung;Roh, In-Soon;Yoon, Byung-Il;Han, Jeong-Hee
    • Journal of Veterinary Clinics
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    • v.26 no.6
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    • pp.556-562
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    • 2009
  • Skin tumors and mammary gland tumors have been shown to be the most common neoplasia in most of the strains of dogs. The risk for tumor development increases significantly with age and the prevalence and distribution are various according to individual tumors. The aim of this study is to classify histopathologically the skin and mammary gland tumors for recent two years, 2005 and 2006. A total of 128 skin and 240 mammary gland samples of dogs were selected that were submitted to National Veterinary Research and Quarantine Service and Kangwon National University from January 1, 2005 to December 31, 2006. The excised tissue were fixed in 10 percent neutral buffered formalin and processed routinely to paraffin wax. Sections were cut at $3{\mu}m$, stained with haematoxylin and eosin. The slides were examined based on the morphological criteria of M. H. Goldschmidt and W. Misdorp under a light microscope. The age of the dogs ranged from 1 to 19 years with a median of 8.7 years. The mean age of the skin and mammary gland tumors was 7.4 and 9.3 years. 47 (12.8%) were males and 259 (70.4%) were female with a male to female ratio of 0.18. Yorkshire terrier and maltese were more susceptible breeds, accounting for 44.3% of skin and mammary gland tumors. In skin tumors, epithelial, adnexal, and mesenchymal origin tumors were 18 (14.1%), 53 (41.4%), and 57 cases (44.5%), repectively. Among the epithelial, adenexal, and mesenchymal origin tumors, basal cell tumor (8.6%), sebaceous adenoma (15.6%), and histiocytoma (25.0%) were predominant in the incidence rate, respectively. In case of mammary gland tumors, 201 (83.8%) were benign and 39 (16.3%) were malignant with a benign to malignant ratio of 5.15. The most frequent mammary gland tumor was benign mixed tumor (35.0%) followed by mammary adenoma-complex type (31.7%).

Annual Reprodutive Cycle of the Jackknife Clams, Solen strictus and Solen gordonis (맛조개, Solen strictus와 붉은맛, Solen gordonis의 생식년주기)

  • CHUNG Ee-Yung;KIM Hyung-Bae;LEE Taek-Yuil
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.19 no.6
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    • pp.563-574
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    • 1986
  • The structure of gonads, gametogenesis and reproductive cycle of the jackknife clams, Solen strictus and Solen gordonis were investigated mainly by histological observation. The first species used were monthly sampled at the coastal area of Dadaepo, Pusan, Korea and Naechodo, Kunsan, Korea for one year from February 1982 to January 1983. The second species were monthly sampled at the sand beach of Dadaepo, Pusan, Korea, from February 1982 to January 1983. Sexualities of Solen strictus and Solen gordonis are dioecious, and these species are oviparous. The gonads are irregularly arranged from the subregion of mid-intestinal gland in visceral cavity to reticular connective tissue of foot. The ovary was composed of a number of small ovarian sacs and the testis was composed of several testicular lobuli which from the tubular structure. Early multiplicating oogonium was about $10{\mu}m$ in diamater. Nucleus and nucleolus, at that time, were distinct in appearance. Each of the early growing oocytes made an egg-stalk, connected to the germinal epithelium of the ovarian sac. A great number of undifferentiated mesenchymal tissue and eosinophilic granular cells are abundantly distributed in the ovarian sacs in the early development stages. With the further development of gonad, these tissue and cells gradually disappeared. Then the undifferentiated mesenchymal tissue and eosinophilic granular cells function as nutritive cells in the formation and development of the early stage germ cells. Mature oocytes were free in the lumen of ovarian sacs and gradually become round or oval. Ripe oocyte was about 80 to $90{\mu}m$ in diameter. With the further development of testis, each of the testicular lobuli formed stratified layers composed of spermatogonia, spermatocytes, spermatids and spermatozoa in groups on the germinal epithelium. After spawning, the gonad gradually degenerated, and disorganized completely. Then new differentiated tissues were rearranged next year. The annual reproductive cycle of those species could be classified into five stages; multiplicative, growing, mature, spent, degenerative and resting stage. It seems that the spawning season is closely related to the water temperature, and the spawning of Solen strictus occurs from June to July at above $20^{\circ}C$ in water temperature. The peak spawning season appeared in June at Dadaepo and in July at Kunsan, The spawning of Solen gordonis occurs from May to June with the peak spawning season in June. Percentages of the first maturity in female of Solen strictus ranging from 5.1-6.0 cm and 7.1-8.0 cm in shell length were $50\%$ and $100\%$, respectively.

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A STUDY ABOUT THE APOPTOSIS OF EPITHELIAL CELLS IN THE FUSING FETAL RAT PALATE (흰쥐 태아 구개융합부위 상피세포의 아포프토시스에 관한 연구)

  • Lee, Hong-Joo;Cha, Kyung-Suk;Lee, Jin-Woo
    • The korean journal of orthodontics
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    • v.31 no.1 s.84
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    • pp.71-83
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    • 2001
  • The purpose of this study was to Prove that the medial edge epithelial cells covering the secondary palatal shelves were removed by apoptosis during palatal fusion. 12 mature female rats (Suprague-Dawley) were mated overnight with male rats and sacrificed on days 15.0, 16.5, 16.75, 17.0 of pregnancy. The embryos were removed from the uterus and the heads were embedded in paraffin. The paraffin blocks were sectioned and the sections were undergone H-E staining for general histologic feature and TdT staining for detection of apoptotic cells. The obtained results were as follows. k. In the section of 16.0 and 16.5 day embryos, the palatal shelves were prior to contact and no apoptotic cells wereobserved in the medial edge epithelium. At the initial contact of Palatal shelves, there was a few apoptotic cells in the fusing epithelium. 2. In the 16.75 day embryos, the samples that epithelial seams did not lost there continuity, apoptotic cells were rarely seen at the midline epithelial seam. In contrast, a lot of apoptotic cells were observed at epithelial triangles and the junction between palatal shelves and nasal septum. 3. In the 16,75 day embryos, the samples that epithelial seams lost their continuity and disrupted to epithelial islands, large number, of apoptotic cells were observed at epithelial islands and epithelial triangles. Some apoptotic cells were also observed at the oral, nasal epithelium near the midline. 4. In the 17.0 day embryos, most of epithelial islands were disappeared and mesenchymal confluence was achieved. Apoptotic cells were rarely observed in the mesenchymal tissue which replaced epithelial islands, but there were some apoptotic cells at the epithelial triangles, oral and nasal epithelium. From the results of the study, it was revealed that medial edge epithelial cells of fusing palate were removed by apoptosis. Apoptotic cells were found mainly in the disappearing midline epithelial seam and the oral and nasal epithelial triangles at some late stages of palatal fusion.

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Radioprotective Effect of Red Ginseng in Irradiated Mice with ${\gamma}$-ray (생쥐에서 홍삼의 감마선조사에 의한 방어효과)

  • Seung, Ka-Yeon;Lee, Heung-Man;Kim, Jong-Sang;Lee, Jun-Haeng
    • Journal of the Korean Society of Radiology
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    • v.4 no.1
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    • pp.31-35
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    • 2010
  • Recently, the incidents of direct or indirect radiation exposure due to increase of use of radiation or radioisotope are on the increase in medical and industrial circles. If cells are irradiated, free radicals are created through biological process, and cells are directly or indirectly damaged. This research intends to explore into the effect of saponin at the level of cell (in vitro) and entity (in vivo), using red ginseng extract "saponin", as radioprotective agent. In the experiment implemented at the level of cell (in vitro), degree of cell activity was measures by adding mouse mesenchymal stem cells "C3H/10T1/2 cells" into red ginseng extract "saponin(0, 0.05, 0.2, and 0.4 g/L)", and then the optimal concentration of saponin influencing cells was calculated, in 24, 48, 72, and 96 hours after gamma irradiation at the optimal concentration of saponin, each cell survival rate was observed through XTT assay. The best time period of cultivation for the optimal activity of C3H/10T1/2 cells was as 48 hours, and the degree of optimal activity was shown at 0.05 g/L. In 48 hours after irradiation of 5 Gy to C3H/10T1/2 cells at 0.05 g/L, the degree of activity of cells increased by 10%. In the experiment implemented at the level of entity (in vivo), red ginseng extract "saponin" at a dose of 100 mg/kg/day was injected into the abdominal cavity of six-week immature mouse for two weeks. Right after the last abdominal injection, total body irradiation of gamma rays was carried out at a dose of 5 Gy and 10 Gy. And after irradiation, the blood sample was taken, and then the number of red corpuscles was counted. In result, the decrement of experimental group treated with red ginseng extract "saponin" was 2.3 times larger than that of control group. In view of the results so far achieved, it was revealed that red ginseng extract "saponin" has a radiation exposure protection effect in the experiment implemented at the level of cell (in vitro). In case of animal experiment, the decrement of number of red corpuscles decreased. Finally, it is necessary to carry out more various researches continuously.

Surface maker and gene expression of human adipose stromal cells growing under human serum. (인체혈청 하에서 배양한 인체지방기질줄기세포의 표면항원 및 유전자 발현)

  • Jun, Eun-Sook;Cho, Hyun-Hwa;Joo, Hye-Joon;Kim, Hoe-Kyu;Bae, Yong-Chan;Jung, Jin-Sup
    • Journal of Life Science
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    • v.17 no.5 s.85
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    • pp.678-686
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    • 2007
  • Human mesenchymal stem cells(hMSC), that have been reported to be present in bone marrow, adipose tissues, dermis, muscles and peripheral blood, have the potential to differentiate along different lineages including those forming bone, cartilage, fat, muscle and neuron. Therefore, hMSC are attractive candidates for cell and gene therapy. The optimal conditions for hMSC expansion require medium supplemented with fetal bovine serum(FBS). Some forms of cell therapy will involve multiple doses, raising a concern over immunological reactions caused by medium-derived FBS proteins. Previously, we have shown that hADSC can be cultured in human serum(HS) during their isolation and expansion, and that they maintain their proliferative capacity and ability for multilineage differentiation and promote engraftment of peripheral blood-derived CD34 cells mobilized from bone marrow in NOD/SCID mice. In this study we determined whether hADSC grown in HS maintain surface markers expression similar with cells grown in FBS during culture expansion and compared gene expression profile by Affymetrix microarray. Flow cytometry analysis showed that HLA-DR, CD117, CD29 and CD44 expression in HS-cultured hADSC during culture expansion were similar with that in FBS-cultured cells. However, the gene expression profile in HS-cultured hADSC was significantly different from that in FBS-cultured cells. Therefore, these data indicated that HS-cultured hADSC should be used in vivo animal study of hADSC transplantation for direct extrapolation of preclinical data into clinical application.

Effect of Prostaglandin F2 Alpha on E-cadherin, N-cadherin and Cell Adhesion in Ovarian Luteal Theca Cells (난소의 황체협막세포에서 E-cadherin, N-cadherin과 세포부착에 미치는 Prostaglandin F2 Alpha의 영향)

  • Lee, Sang-Hee;Jung, Bae Dong;Lee, Seunghyung
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.3
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    • pp.360-369
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    • 2019
  • Cadherins are essential transmembrane proteins that promote cell-cell adhesion and maintain the corpus luteum structure in the ovary. This study examined the influence of prostaglandin F2 alpha ($PGF2{\alpha}$) on E-cadherin, N-cadherin, and adhesion in luteal theca cells (LTCs). The luteal cells were isolated from the mid-phase corpus luteum, and the LTCs were cultured separately from the luteal heterogeneous cells according to the morphology of the mesenchymal cells and to determine if steroidogenic and endothelial cells of LTCs, 3beta-hydroxysteroid dehydrogenase ($3{\beta}$-HSD), and vascular endothelial growth factor receptor 2 (VEGFR2) mRNA were used. The LTCs were then incubated in the culture medium supplemented with 0.01, 0.1, and 1.0 mM $PGF2{\alpha}$ for 24 h, and the E-cadherin and N-cadherin proteins in the LTCs were detected by confocal laser scanning microscopy. The results revealed $3{\beta}$-HSD mRNA expression in the LTC but no VEGF2R mRNA expression. The E-cadherin and N-cadherin proteins of the LTCs were damaged in the 0.01, 0.1, and 1.0 mM $PGF2{\alpha}$ treatment groups, and the expression of the N-cadherin protein was reduced significantly in 0.01 mM $PGF2{\alpha}$ compared to the 0 mM $PGF2{\alpha}$ treatment groups (P<0.05). In addition, the number of attached LTCs were significantly lower in the 0.01 mM $PGF2{\alpha}$ treatment group than in the 0 mM $PGF2{\alpha}$ treatment group (P<0.05). In conclusion, $PGF2{\alpha}$ affected the disruption of cadherin proteins and cell adhesion in LTCs. These results may help better understand the cadherin and adhesion mechanism during corpus luteum regression in the ovary.

Alteration of MicroRNAs Targeted Integrins by PD-MSCs Transplantation Is Involved in Hepatic Regeneration in a Rat Model with BDL (담관결찰 쥐 모델에서 태반유래중간엽줄기세포 이식에 의한 miRNA 표적 인테그린 변화의 간재생 효과)

  • Park, Sohae
    • Journal of Life Science
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    • v.31 no.8
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    • pp.710-718
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    • 2021
  • Placenta-derived mesenchymal stem cells (PD-MSCs) are promising candidates for cell-based therapy in regenerative medicine. The migration and homing potential of PD-MSCs to injured sites is a critical property of MSC engraftment. MicroRNAs (miRNAs) have recently been shown to regulate the critical functions of MSCs, such as proliferation, survival, and migration. The objective of the present study was to identify the miRNA and target genes involved in PD-MSCs homing in a bile duct ligation (BDL) rat model. We selected candidate miRNAs targeting genes for PD-MSCs homing based on microarray analysis. PD-MSC engraftment in BDL-injured rat liver was identified by immunofluorescence assay and human-specific Alu gene expression by quantitative real-time polymerase chain reaction (qRT-PCR) one week after transplantation. Compared with migrated naïve PD-MSCs under hypoxic and normoxic conditions (Hyp/Nor), the transplanted group with PD-MSCs (Tx) showed distinct differences in miRNA expressions in BDL-injured rat liver. We also validated the miRNAs and their target genes for PD-MSCs homing. The expressions of integrin α4 (ITGA4) and integrin α5 (ITGA5) target genes for miR-199a-5p and miR-148a-3p were significantly upregulated in the Tx group (p<0.05). In addition, integrin β1 (ITGB1) and integrin β8 (ITGB8) were upregulated by suppressing miR-183-5p and miR-145-5p, respectively. These results demonstrated that PD-MSCs regulate miRNA expression related to the integrin family for their homing effects on the BDL-injured rat liver. The findings further suggest that miRNA-mediated regulation of the integrin family contributes to the therapeutic efficacy of PD-MSCs in the rat hepatic fibrosis model by BDL.