• Title/Summary/Keyword: PCR 동정

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Clinical Significance of Low-colony Count Scotochromogen Nontuberculous Mycobacteria (균집락수가 적은 암색소성 비결핵항산균 배양의 임상적 의미)

  • Lee, Jung Yeon;Kim, Mi-Na;Chung, Hee-Jung;Jun, Kyung Ran;Choi, Hee Jin;Lee, Hyeyoung;Joung, Eun Young;Oh, Yeon-Mok;Lee, Sang Do;Kim, Woo Sung;Kim, Dong Soon;Kim, Won Dong;Shim, Tae Sun
    • Tuberculosis and Respiratory Diseases
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    • v.59 no.1
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    • pp.39-46
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    • 2005
  • Background : Even though it has been suggested that low-colony, scotochromogen nontuberculous mycobacteria (NTM) are usually contaminants and not true pathogens, evidence for this hypothesis has not been provided. This study investigated the colony characteristics, organism identification, and clinical significance of low-colony scotochromogen. Methods : The laboratory cultured 6,898 respiratory clinical specimens for an examination of mycobacteria over a three-month period. A low-colony count was arbitrarily defined as ${\leq}20$ colonies. This study analyzed the recovery rate of the mycobacteria, the number of colonies and their gross characteristics, and their clinical significance. PCR-restriction fragment length polymorphism analysis was carried out to identify the NTM species. NTM pulmonary disease was defined according to the American Thoracic Society. Results : A total of 6,898 respiratory specimens for mycobacterium were cultured. Of these, 263 (3.8%) grew NTM, and 382 (5.5%) grew M. tuberculosis. Of the 263 cultured NTM specimens, 124 (47.1%) were scotochromogens. The smear-positive rate was significantly lower in these scotochromogens (4.8%) than in the non-scotochromogens (23.7%) (p<0.05). The most common isolates were M. gordonae (83/102, 81.4%) in the scotochromogens, and MAC (52/121, 43.0%) in the non-scotochromogens. Even though three out of 113 patients with a low-colony scotochromogen has been diagnosed with NTM pulmonary disease, the isolated scotochromogen was not considered to be the cause of the NTM disease but was just a contaminant. Conclusion : In this study, the most common isolate of a low-colony count scotochromogen was M. gordonae, which appeared to be contaminants and not true pathogens. Greater efforts in the quality control of a mycobacterium laboratory are needed in cases where there is a high recovery rate of low-colony count scotochromogen.

Characters of motile aeromonads isolated from imported ornamental fish (수입 관상어에서 분리한 motile aeromonads의 특성)

  • Jin, Se-yun;Ko, Chang-yong;Lee, Ye-ji;Jung, Yun-hee;Ju, Seong-cheol;Kim, Eunheui
    • Journal of fish pathology
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    • v.30 no.2
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    • pp.89-96
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    • 2017
  • The majority of freshwater ornamental fish are imported and distributed domestically, causing high risk of exposure to exotic pathogens and drug resistant bacteria in Korea. Aeromonas hydrophila is known as a common species of fresh water bacteria and opportunistic fish pathogen, as well as a species causing zoonotic infection. In this study, we isolated motile aeromonads from various imported freshwater ornamental fish and studied the characters of the isolates. Imported freshwater ornamental fish were purchased on day 1 after the fish were deposited in the aquarium. Bacteria were isolated from the liver, kidney and spleen of fish using 0.5% NaCl containing tryptic soy agar medium. Bacteria were grouped on the basis of their morphological characteristics. The colonies with clear zone on starch-ampicillin agar (SA agar) were tentatively identified as Aeromonas spp. Two hundred and twenty-six strains, about 70% of total isolates were assumed to be Aeromonas spp. Nine isolates were further identified based on the result of the API 20E test and PCR using primers specific for A. hydrophila 16S rRNA gene. The isolates were identified as A. hydrophila and the API 20E test showed differences in trisodium citrate, D-sucrose, D-melibiose, amygdalin and L-arabinose availability between the nine isolates and standard A. hydrophila. The susceptibilities of the isolated bacteria to 10 antibacterial agents were confirmed by the disk diffusion method. Isolated strains were found to be resistant to amoxicillin and ampicillin and sensitive to florfenicol. However, 7 isolates showed multiple drug resistances to erythromycin, oxytetracycline, nalidixic acid etc. Pathogenicity of the isolates was determined by the artificial challenge test on goldfish (Carassius auratus). Three isolates caused 60 ~ 80% mortality in goldfish within 5 days after the initiation of challenge. These results indicate that multiple drug resistant, highly pathogenic and exotic A. hydrophila can spread to domestic aquarium and the preventive treatment of fish before sale is necessary.

Identification of Genes Connected with the Sensitivity to 5-FU and Cisplatin in Squamous Cell Carcinoma Cell Lines (편평세포암 세포주에서 5-FU와 Cisplatin에의 감수성과 관련된 유전자의 동정)

  • Choi, Na-Young;Kim, Ok-Joon;Lee, Geum-Sug;Kim, Byung-Gook;Kim, Jae-Hyeong;Jang, Youn-Young;Lim, Won-Bong;Chong, Min-A;Choi, Hong-Ran
    • Journal of Oral Medicine and Pain
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    • v.30 no.3
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    • pp.287-300
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    • 2005
  • Squamous cell carcinoma (SCC) in head and neck show a variability in the response to chemotherapy, even when it present with similar histological tumor type, grade, and clinical stage. The purpose of present study it to identify predictive bio-marker for the sensitivity or resistance to conventional chemotherapeutic agents, 5-fluorouracil (5-FU) and Cisplatin Oral cancer cell lines were used in present study. MTT assay was performed to evaluate the sensitivity and/or resistance to 5-FU and Cisplatin. And RT-PCR was carried out for evaluation of the mRNA expressions of various genes associated with mutation, inflammation (COX pathway), cell cycle, senescence and extracellular matrix (ECM). The molecules which are correlated with the sensitivity to 5-FU are XPA, XPC, OGG, APEX, COX-2, PPAR, Cyclin E, Cyclin B1, CDC2, hTERT, hTR, TIMP-3, TIMP-4 and HSP47. And the molecules are correlated with the sensitivity to Cisplatin are COX-1, iNOS, eNOS, PCNA, collagen 1 and MMP-9. Taken together, when choosing the appropriate chemotherpeutic agents for patients, considering the molecules which are correlated or reversely correlated is helpful to choose the resonable agents for cancer patients.

Microbial and Pathogenic Contamination of Ready-to-eat Fresh Vegetables in Korea (한국에 유통중인 신선편이 채소류의 미생물 품질 및 병원성 세균의 오염도 조사)

  • Bae, Young-Min;Hong, Yu-Jin;Kang, Dong-Hyun;Heu, Sung-Gi;Lee, Sun-Young
    • Korean Journal of Food Science and Technology
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    • v.43 no.2
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    • pp.161-168
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    • 2011
  • The purpose of this study was to evaluate microbiological contamination of fresh vegetables in Korea. Twenty types of vegetables were tested for total aerobic bacteria, coliforms, Escherichia coli, yeast and mold, and pathogenic bacteria such as Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus, Salmonella, E. coli O157:H7, Cronobacter sakazakii, Shigella, and Campylobacter. Levels of total aerobic bacteria and coliform on 20 vegetables were between 3.74 and 8.04 log CFU/g, and 0.16 and 5.02 log CFU/g, respectively. The highest contamination levels of total aerobic bacteria were observed on water dropwort, sprouts, mungbean sprout, and ballflower root. There was no significant difference (p>0.05) in microbial contamination levels of total aerobic count, coliform, E. coli, yeast and mold between organic and nonorganic vegetables. When isolation methods using selective agars were applied, L. monocytogenes, B. cereus, Salmonella and Campylobacter were isolated from some fresh vegetable samples. Results of API kit tests showed that L. monocytogenes was identified on Chinese cabbage, cucumber, soybean sprouts, and iceberg lettuce while Salmonella was identified on Korean leek. Furthermore, Campylobacter jejuni was also identified in more than 50 of the 100 samples. However, when positive samples from API kit were tested for real-time PCR or 16S rRNA sequencing method, only B. cereus from perilla leaf, carrot, water dropwort, and sprouts showed positive results. These results indicate that selective agar and API kit detection methods might result in false positive results for some pathogens. Therefore, studies need to improve isolation or confirmation methods for such pathogens.

Prevalence of CTX-M-type Extended-Spectrum $\beta$-Lactamases Producing Escherichia coli and Klebsieilla pneumoniae Isolates in General Hospitals in 2005 (임상에서 분리된 CTX-M형 Extended-Spectrum $\beta$-Lactamases를 생산하는 Escherichia coli와 Klebsiella pneumoniae의 유행)

  • Kim, Yun-Tae;Kim, Tae-Un
    • Microbiology and Biotechnology Letters
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    • v.34 no.4
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    • pp.342-351
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    • 2006
  • The aim of this study was to survey susceptibilities of Escherichia coli and Klebsiella pneumoniae isolates against cefotaxime and to determine the prevalences of CTX-M type extended-spectrum $\beta$-lactamases (ESBLs) producing E. coli and K. pneumoniae in Korea. During the period of February to July, 2005, 153 E. coli and 52 K. neumoniae isolates were collected from 2 hospitals in Busan. Antimicrobial susceptibilities to cefotaxime were tested by the disk diffusion method. ESBL production of E. coli and K. pneumoniae was determined by the double disk synergy test. MICs of $\beta$-lactam antibiotics were determined by the agar dilution method. Blac$_{CTX-M}$ genes of the organism were detected by PCR. Among 153 isolates of E. coli and 52 isolates of K. neumoniae, 27 (17.6%) and 25 (48.0%) were intermediate or resistant to cefotaxime, respectively. Twenty-three (15.0%) isolates out of 153 E. coli and 13 (25.0%) out of 52 K. neumoniae isolates showed positive results for ESBL by the double disk synergy test. Twenty isolates out of 23 ESBL producing E. coli and 12 out of 13 ESBL producing K. neumoniae isolates harbored biacTx-M gene,11 of ESBL producing E. coli and 12 of ESBL producing K. neuinoniae isolates harbored bla$_{CTX-M}$ gene, 11 of the ESBL producing E. coli and 2 of ESBL producing K. neumoniae isolates harbored bla$_{TEM}$ gene, and 1 of the ESBL producing E. coli and 12 of ESBL producing K. neumoniae isolates harbored bla$_{SHV}$ gene. E. coli and K. neumoniae isolates producing CTX-M-type ESBLs were not uncommon in Korea. It is thought that continuous survey are necessary for inspecting the spread and novel variants of CTX-M-type ESBL genes. Further me]'e investigation and research on ESBL producing strains are needed in order to prevent the spread of resistant bacteria.

Immunomodulatory Effects of β-sitosterol and Daucosterol Isolated from Dioscorea batatas on LPS-stimulated RAW 264.7 and TK-1 Cells (산약에서 분리한 β-sitosterol과 daucosterol의 RAW 264.7 세포와 TK-1 세포에서의 면역 활성 조절 효능)

  • Park, Min-Kyung;Cho, Sehee;Ahn, Tae-Kyu;Kim, Do-Hyun;Kim, So-Yeon;Lee, Jin-Wook;Kim, Jee-In;Seo, Eul-Won;Son, Kun-Ho;Lim, Jae-Hwan
    • Journal of Life Science
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    • v.30 no.4
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    • pp.359-369
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    • 2020
  • Although many studies on immune modulatory materials have used RAW 264.7 cells, few have used T cell-derived TK-1 cell lines. Moreover, although some studies have investigated the efficacy of plant-derived β-sitosterol, few have examined the immunomodulatory activity of its analogue, daucosterol. In this study, β-sitosterol and daucosterol were isolated from D. batatas and identified by nuclear magnetic resonance spectroscopy. To evaluate the immune-enhancing or inhibitory effects of the isolated phytosterols, the expression levels of the inflammatory response genes COX-2, TNF-α, IL-6, and iNOS were analyzed by RT-PCR. The relative expression levels of TNF-α and iNOS in RAW 264.7 cells were increased more than threefold with β-sitosterol treatment comparing to those of untreated control. In the case of TK-1 cells, the expression level of TNF-α was decreased and the expression level of iNOS was increased in a β-sitosterol concentration-dependent manner. The expression levels of COX-2, TNF-α, and IL-6 increased by approximately 0.7-1.2 times in RAW 264.7 cells treated with daucosterol compared to those of untreated control, but iNOS expression decreased by 0.8-0.18 times. In the case of daucosterol-treated TK-1 cells, the expression levels of TNF-α, IL-6, and iNOS were markedly reduced from those of TK-1 cells treated only with lipopolysaccaride. As a conclusion, β-sitosterol treatment increased TNF-α and iNOS expression levels in RAW 264.7 cells, thus exerting an immune- boosting effect. However, in TK-1 cells, iNOS expression increased while TNF-α expression decreased, indicating an immunosuppressive activity of β-sitosterol. Daucosterol appears to exert an immunosuppressive effect in both macrophages and T cell lines by inhibiting iNOS expression in RAW 264.7 cells and greatly inhibiting the expression of TNF-α, IL-6, and iNOS in TK-1 cells.

Isolation of Mycoplasma pneumoniae and Antimicrobial Susceptibilities of the Isolates(III) (Mycoplasma pneumoniae의 분리 및 항생제 감수성 검사(III))

  • Chang Myung-Woong;Kim Kwang-Hyuk;Park In-Dal;Song Gap-Young;Kim Sung-Won;Lee Eun-young;Kim Moon-Chan;Cho Myung-Hoon;Kim Kyu-Earn;Choi Choong-Eon;Park Seon Yeong;Jo Hyeon Jang
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.479-485
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    • 2005
  • The 994 throat swabs obtained from 688 adults and 306 children patients with respiratory diseases were examined for Mycoplasma pneumoniae infection by culture method. Antimicrobial susceptibilities of the resulting 123 M. pneumoniae isolates were evaluated by testing minimum inhibitory concentrations (MICs) of erythromycin, minocycline, tetracycline, josamycin, sparfloxacin, ofloxacin, and ciprofloxacin by a broth micro-dilution method. The erythromycin resistant strains of M. pneumoniae was determined above $1.0{\mu}g/ml$ of MIC for erythromycin. The erythromycin resistant strains of M. pneumoniae was confirmed resistant gene mutation of the portions of genes 23S rRNA (domain II and V), and ribosomal protein 14 and L22 by PCR amplified and their nucleotide sequenses were compared to those of the susceptible strain M129. The isolation rate of M. pneumoniae was $12.9\%$ (89/688) for the adults and $11.1\%$ (34/306) for the children. The $MICs_{90}$ of the M. pneumoniae isolates were $0.12{\mu}g/ml$ for minocycline, $0.25{\mu}g/ml$ for sparfloxacin, $0.5{\mu}g/ml$ for ciprofloxacin, ofloxacin, and tetracycline, respectively, and $2.0{\mu}g/ml$ for josamycin and erythromycin, respectively. The isolation rate of erythromycin resistant M. pneumoniae from patients was $49.4\%\;(44/89)$ for the adults, $47.1\%\;(16/34)$ for children, and $48.8\%\;(60/123)$ for the total. No mutation could be detected in the ribosomal protein L22 region, but all strains were mutated in the ribosomal protein L4 as two point mutation M144V. Two point mutations in domain V of 23S rRNA were selected in the presense of erythromycin resistant M. pneumoniae isolates, such as one strain was G2057C mutant, two strains were A2059C mutants, three strains were C2611G mutants, four strains were A2058C mutants, five strains were A2058T mutants, twenty strains were A2059G mutants, and twenty-five strains were A2058G mutants, respectively. These results show that erythromycin was not the most active compound against M. pneumoniae infection in Korea and clinical studies of macrolides in human patients are demanded.

Assessment of Microbiological Quality for Raw Materials and Cooked Foods in Elementary School Food Establishment (초등학교에 공급되는 급식용 식재료 및 조리식품의 미생물학적 품질평가)

  • Shin, Weon-Sun;Hong, Wan-Soo;Lee, Kyung-Eun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.3
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    • pp.379-389
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    • 2008
  • This study was conducted to assess the microbiological quality of raw and cooked foods served in the elementary school food service. Raw and cooked food samples were collected from 11 selected elementary schools in both June to July and September to October of 2005. Petrifilm plates were used to determine (in duplicate) total aerobic colony counts (PAC), Enterobacteriaceae (PE), coliform counts (PCC), and E. coli counts (PEC). Heavy contamination of Enterobacteriaceae (from 0.08 to 7.40 log CFU/g) and total coliform (0.50 to 6.52 log CFU/g) were observed in raw materials and cooked foods. Escherichia coli (E. coli) were detected in the sample of currant tomato (3.70 log CFU/g), sesame leaf (3.59 log CFU/g), dropwort (0.20 log CFU/g), crown daisy (3.15 log CFU/g), parsley (3.00 log CFU/g), peeled green onion (1.74 log CFU/g), frozen pork (0.65 log CFU/g), frozen beef (0.20 or 1.50 log CFU/g), chicken (1.78 log CFU/g), and young radish leaf seasoned with soybean paste (1.24 log CFU/g). Multiplex PCR system was used to determine the food-borne pathogens: Salmonella spp., Bacillus cereus (B. cereus), E. coli O157:H7, Staphylococcus aureus, Listeria monocytogenes (L. monocytogenes), Vibrio parahaemolyticus, Campylobacter jejuni (C. jejuni), Shigella spp., B. cereus was detected in 19 samples of raw materials and 8 samples of cooked foods. With regard to quantitative analysis, B. cereus counts exceeded 5.46, 3.48 and 1.79 log CFU/g in sesame leaf, peeled green onion and seasoned mungbean jelly, respectively. E. coli O157:H7 was detected on 2 samples of frozen beefs, and its biochemical characteristics of one beef sample was confirmed with API 20E kit (93.7%). L. monocytogenes was detected in fried rice paper dumpling, but the presumptive colonies were not detected onto the conventional plate. C. jejuni was detected in peeled & washed onion.

Carriage Rates and Serogroups of Neisseria meningitides in Children Attending Day Care Centers (유아원에 다니는 소아의 구인강 내 수막구균 보균율과 혈청군)

  • Kim, Nam Hee;Lee, Jina;Lee, Jung Won;Lee, Soo Young;Choi, Eun Hwa;Kim, Kyung Hyo;Kim, Eui Chong;Kang, Jin Han;Lee, Hoan Jong
    • Pediatric Infection and Vaccine
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    • v.16 no.1
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    • pp.31-39
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    • 2009
  • Purpose : Neisseria meningitides is one of the most common causative pathogens of bacteremia and meningitis. Recently protein-conjugated vaccines have been developed and included in the routine vaccination schedule in a few countries. In Korea, carriage rates of N. meningitides among healthy adults have been reported. However, systematic data for childhood carriage rates are not available. This study was performed to evaluate the carriage rates of N. meningitides and the serotype distribution among healthy children attending day care centers. Methods : During the period of January through May 2005, nasopharyngeal swabs and culture were obtained from 904 children attending 13 different day care centers located in Seoul and Gyeonggi Province. The Vitek NHI card was used to identify N. meningitides and the crgA gene was detected via polymerase chain reaction (PCR). Serotype determination was performed by agglutination test using N. meningitides antisera to serotypes A, B, C, D, 29E, W135, X, Y, and Z. PCR for detection of the org2 and saiD gene confirmed serotypes A, B, C, W135, and Y. Results : The mean age among 904 children was 4.5 years; 6.5% (59/904) were children <2 years old, 53.8% (486/904) were 2-5 years old, and 39.7% (359/904) were >5 years old; 52.0% (468/904) were male. N. meningitides was isolated from only 7 children attending 5 different day care centers and the overall carriage rate of N. meningitides was 0.8%. The detected serotypes of N. meningitides were serotype A (n=2), C (n=2), and Y (n=3). Conclusion : The carriage rate of N. meningitides among healthy children attending day care centers was very low in Korea and the detected serotypes were A, C, and Y.

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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.