• Asian-Australasian Journal of Animal Sciences
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• 제21권11호
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• pp.1544-1550
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• 2008

The retinoids (vitamin A and its derivatives) play a critical role in vision, growth, reproduction, cell differentiation and embryonic development. Using the IMpRH panel, porcine cellular retinol binding protein genes 5 and 7 (RBP5 and RBP7) were assigned to porcine chromosomes 5 and 6, respectively. The complete coding sequences (CDS) of the RBP5 and RBP7 genes were amplified using the reverse transcriptase polymerase chain reaction (RT-PCR) method, and the deduced amino acid sequences of both genes were compared to human corresponding proteins. The mRNA distributions of the two genes in adult Wuzhishan pig tissues (lung, skeletal muscle, spleen, heart, stomach, large intestine, lymph node, small intestine, liver, brain, kidney and fat) were examined. A total of nine single nucleotide polymorphisms (SNPs) were identified in two genes. Three of these SNPs were analyzed using the polymerase chain reaction-restriction-fragment length polymorphism (PCR-RFLP) method in Laiwu, Wuzhishan, Guizhou, Bama, Tongcheng, Yorkshire and Landrace pig breeds. Association analysis of genotypes of these SNP loci with economic traits was done in our experimental populations. Significant associations of different genotypes of $RBP5-A/G^{63}$, $RBP5-A/G^{517}$ and $RPB5-T/C^{intron1-90}$ loci with traits including maximum carcass length (LM), minimum carcass length (LN), marbling score (MS), back fat thickness at shoulder (SBF), meat color score (MCS) and hematocrit (HCT) were detected. These SNPs may be useful as genetic markers in genetic improvement for porcine production.

• Asian-Australasian Journal of Animal Sciences
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• 제33권1호
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• pp.4-11
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• 2020

Objective: Puerarin has the potential of regulating the differentiation of preadipocytes, but its mechanism of action has not yet been elucidated. Adipocytes found in adipose tissue, the main endocrine organ, are the main sites of lipid deposition, and are widely used as a cell model in the study of in vitro fat deposition. This study aimed to investigate the effects of puerarin on adipogenesis in vitro. Methods: Puerarin was added to the culture medium during the process of adipogenesis. The proliferation and differentiation of bovine preadipocytes was measured through cell viability and staining with oil red O. The content of triacylglycerol was measured using a triglyceride assay kit. The mRNA and protein expression levels of adipogenic genes, peroxisome proliferator-activated receptor-γ (PPARγ) and CCAAT/enhancer-binding protein-α, were measured using quantitative real-time polymerase chain reaction and western blotting, respectively. Results: The addition of puerarin significantly increased adipogenesis of bovine preadipocytes and enhanced the mRNA and protein level expression of PPARγ (p<0.01). The expression of P-Akt increased after adipogenic hormonal induction, whereas puerarin significantly increased PPARγ expression by promoting the Akt signaling component, P-Akt. The mechanism of adipogenesis was found to be related to the phosphorylation level of Ser473, which may activate the downstream signaling of the Akt pathway. Conclusion: Puerarin was able to promote the differentiation of preadipocytes and improve fat deposition in cattle. The mechanism of adipogenesis was found to be related to the phosphorylation level of Ser473.

• 생명과학회지
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• 제20권10호
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• pp.1549-1555
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• 2010

Streptococcus suis는 사람과 돼지에서 다양한 감염을 유발하는 병원체이다. 2004년부터 2009년까지 경북지방 양돈장에서 병성감정 의뢰한 돼지 가검물을 대상으로 S. suis를 분리하여 생화학적 성상, 약제감수성 시험 및 PCR을 이용한 동정을 실시한 결과는 다음과 같다. 40개 농장에서 분리한 61주의 S. suis에 대하여 생화학적 성상시험 결과 모두 S. suis로 동정되었으며, VP 음성, hippurate, esculin, arginine decarboxylase 양성, lactose 분해능이 있는 것으로 나타났다. Trehalose와 starch 분해능은 비교적 높게 나타났으며 생화학적 성상에 따른 phenotype은 11형으로 나누어졌다. 항생제 감수성 시험 결과 대부분의 균주가 amoxicillin/clavulanic acid, ampicillin, cephalothin, cefoperazone, florfenicol에 높은 감수성을 나타내었지만 amikacin, kanamycin, neomycin, streptomycin, erythromycin, clindamycin, tetracycline에는 높은 내성을 나타내었다. 16S-rRNA 특이유전자 부위를 이용한 PCR 결과 공시균주 모두 304 bp에서 증폭산물이 관찰되었다. 이들 결과는 국내에서 사육되고 있는 돼지에서 S. suis 감염을 예방하는데 도움이 될 것으로 기대된다.

• Childhood Kidney Diseases
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• 제12권1호
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• pp.11-22
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• 2008

Telomeres consist of tandem guanine-thymine(G-T) repeats in most eukaryotic chromosomes. Human telomeres are predominantly linear, double stranded DNA as they ended in 30-200 nucleotides(bases,b) 3'-overhangs. In DNA replication, removal of the terminal RNA primer from the lagging strand results in a 3'-overhang of uncopied DNA. This is because of bidirectional DNA replication and specificity of unidirectional DNA polymerase. After the replication, parental and daughter DNA strands have unequal lengths due to a combination of the end-replication problem and end-processing events. The gradual chromosome shortening is observed in most somatic cells and eventually leads to cellular senescence. Telomere shortening could be a molecular clock that signals the replicative senescence. The shortening of telomeric ends of human chromosomes, leading to sudden growth arrest, triggers DNA instability as biological switches. In addition, telomere dysfunction may cause chronic allograft nephropathy or kidney cancers. The renal cell carcinoma(RCC) in women may be less aggressive and have less genomic instability than in man. Younger patients with telomere dysfunction are at a higher risk for RCC than older patients. Thus, telomeres maintain the integrity of the genome and are involved in cellular aging and cancer. By studying the telomeric DNA, we may characterize the genetic determinants in diseases and discover the tools in molecular medicine.

• Asian Pacific Journal of Cancer Prevention
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• 제13권11호
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• pp.5799-5804
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• 2012

Family with sequence similarity 189, member B (FAM189B), alias COTE1, a putative oncogene selected by microarray, for the first time was here found to be significantly up-regulated in hepatocellular carcinoma (HCC) specimens and HCC cell lines. mRNA expression of COTE1 in HCC samples and cell lines was detected by reverse transcription-polymerase chain reaction (RT-PCR) and real-time PCR, while protein expression of COTE1 in HCC tissues was assessed by immunohistochemistry. In addition, invasion of HCC cells was observed after overexpressing or silencing COTE1. In the total of 48 paired HCC specimens, compared with the adjacent non-cancer tissues, the expression of COTE1 was up-regulated in 31 (p<0.01). In HCC cell lines, COTE1 expression was significantly higher than in normal human adult liver (p<0.01). Overexpression of COTE1 enhanced HCC-derived LM6 and MHCC-L cellular invasion in vitro. In contrast, COTE1 knockdown via RNAi markedly suppressed these phenotypes, as documented in LM3 and MHCC-H HCC cells. Mechanistic analyses indicated that COTE1 could physically associate with WW domain oxidoreductase (WWOX), a tumor suppressor. COTE1 may be closely correlated with invasion of hepatocellular carcinoma (HCC) cells and thus may serve as an effective target for gene therapy.

• Asian Pacific Journal of Cancer Prevention
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• 제17권11호
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• pp.4985-4989
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• 2016

Long non-coding RNAs (lncRNAs) are a novel class of non-protein coding RNAs that are involved in a wide variety of biological processes. There are limited data regarding the impact of lnc-LAMC2-1:1 rs2147578 as well as CASC8 rs10505477 T>C polymorphisms on cancer development. Here we examined for the first time whether rs2147578 and rs10505477 polymorphisms are associated with childhood acute lymphoblastic leukemia (ALL) in a total of 110 cases and 120 healthy controls. Genotyping was achieved by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The rs2147578 variant increased the risk of ALL in codominant (OR=4.33, 95%CI=2.00-9.37, p<0.0001, CG vs CC, and OR=5.81, 95%CI=2.30-14.69, p=0.0002, GG vs CC), dominant (OR=4.63, 95%CI=2.18-9.86, p<0.0001, CG+GG vs CC), overdominant (OR=1.74, 95%CI=1.02-2.97, p=0.0444, CG vs CC+GG) and allele (OR=1.91, 95%CI=1.32-2.77, p=0.0008, G vs C) inheritance models tested. No significant association was found between the CASC8 rs10505477 T>C variant and risk of childhood ALL. In conclusion, the present study revealed that the lnc-LAMC2-1:1 rs2147578 polymorphism may be a risk factor for developing childhood ALL. Further studies with larger sample sizes with different ethnicities are now required to confirm our findings.

• 한국균학회지
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• 제44권1호
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• pp.1-7
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• 2016

우리나라 토양에서 효모 종 분포특성을 조사하기 위해 먼저 대전광역시를 포함하는 충청남도 전 지역의 밭 토양 61점을 2015년 6월부터 8월까지 수집하여 97균주 20종의 야생효모들을 분리하였고, polymerase chain reaction (PCR)을 통해 internal transcribed spacer (ITS) 부위와 26S rDNA의 D1/D2 부위의 염기서열 상동성 비교법을 이용하여 종을 동정하였다. 이들 가운데 Cryptococcus podzolicus가 11균주를 포함하는 Cryptococcus 속 균이 전체 분리 균주 중 16균주(16%)로 가장 많이 분리되었고, 다음으로 Debaryomyces hansenii와 Trichorsporon asahii도 각각 6주씩 분리되었다.

• Journal of Chest Surgery
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• 제40권4호
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• pp.264-272
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• 2007

배경: 혈관내피 성장인자(vascular endothelial growth factor, VEGF)는 혈관평활근세포(vascular smooth muscle cell)의 증식과 이동을 촉진함으로써 혈관신생에 중요한 역할을 한다. 당뇨병은 VEGF의 발현과 연관되어 정상 혈당상태에서 보다 세포의 증식을 더욱 촉진시킨다. 당뇨병쥐에서 VEGF 수용체의 선택적 차단이 손상된 혈관에서 신내막 형성과 혈관평활근세포의 이동에 미치는 영향에 대해 알아보고자 했다. 대상 뜻 방법: 당뇨병 쥐의 경동맥 풍선손상 모델에서 위약을 투여하거나, 혈관내피 성장 인자 수용체-1(VEGFR-1)에 선택적으로 작용하는 항-Flt-1 펩타이드(anti-Flt-1 peptide; Gly-Asn-Gln-Trp-Phe-Ile)를 풍선손상 2일 전부터 0.5mg/kg의 용량으로 2주간 매일 투여한 군으로 나누어 Hematoxylin-Eosin 염색을 하여 신내막의 형성정도와 혈관내강의 협착정도를 비교하였으며, proliferative cell nuclear antigen (PCNA)에 대한 면역조직화학염색법을 시행하여 세포의 증식정도를 관찰하였다. 혈관평활근세포를 고혈당환경에서 배양하고 transwell assay를 시행하여 혈관평활근세포의 이동 정도를 측정하였다. 고혈당 환경에서 자라고 있는 혈관평활근세포에 50ng/mL의 VEGF를 단독 또는 3ug/mL의 항-Flt-1 펩타이드와 함께 처리하고 일정시간이 지난 후 matrigel filter를 통과한 세포를 세어 아무런 처치를 받지 않은 세포가 이동한 정도와 비교하였다. 또한, 혈관평활근세포에 세포이동 정도 측정 시와 같은 처리를 한 후, RNA를 분리하고 reverse transcription-polymerase chain reaction (RT-PCR)을 시행하여 기질금속단백분해효소(matrix metalloprotenase, MMP)의 발현 정도를 관찰하였다. 결과: 신내막의 면적은 위약 투여 쥐는 $0.24{\pm}0.03 mm^2$이었으나, 항-Flt-1 펩타이드의 처리에 의해 $0.15{\pm}0.04 mm^2$로 유의하게 감소하였으며(p<0.01), 신내막 형성에 따른 내강의 협착 정도도 위약 투여 쥐는 $61.85{\pm}5.11%$, 항-Flt-1 펩타이드 투여 쥐는 $36.03{\pm}3.78%$로 항-Flt-1 펩타이드 투여에 의하여 유의하게 감소하였다(p<0.01). 신내막의 전체 세포수에 대한 PCNA(+)인 세포를 백분율로 구하였으며, 위약 투여 쥐와 항- Flt-1 펩타이드 투여 쥐에서 각각 $52.82{\pm}4.20%,\;38.11{\pm}6.89%$로 나타나 항-Flt-1 펩타이드를 투여한 쥐에서 PCNA(+)인 세포가 유의하게 적음을 보이고 있다(p<0.05). 혈관평활근세포의 이동 정도 측정에서는 항-Flt-1 펩타이드 처리에 의하여 VEGF에 의한 혈관평활근 세포의 이동이 유의하게 감소하였다(p<0.01). 또한, 항-Flt-1 펩타이드 처리에 의하여 VEGF에 의한 MMP-3와 MMP-9 mRNA의 발현 증가가 억제되었다. 결론: 항-Flt-1 펩타이드는 당뇨병쥐의 경동맥손상모델에서 신내막 형성을 억제하였으며, 고혈당 환경에서 배양된 혈관평활근세포의 이동과, MMP-3와 MMP-9의 활성을 억제하였다.

• 생명과학회지
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• 제25권8호
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• pp.880-888
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• 2015

회향은 미나리과에 속하는 다년생식물인 Foeniculum vulgare Mill.의 성숙한 과실로서 항염, 진통, 피부노화방지 등의 효과를 가지고 있어 다양한 질환에 우수한 치료효과를 나타내는 것으로 알려져 왔다. 따라서 본 연구에서는 기능성이 우수한 회향 열매 추출물을 이용하여 피부 질환 치료제 및 피부장벽 기능 개선 소재로서의 적용가능성을 확인하였다. 이를 위해 각질형성세포주인 HaCaT에 회향 열매 메탄올 추출물과 그 분획물(hexane, methyl chloride, ethyl acetate, butanol)을 처리한 후, involucrin, loricrin, filaggrin의 발현과 hyaluronic acid의 생성 및 β-defensin -1, -2, -3, LL-37의 mRNA 발현을 측정하였다. 그 결과butanol 분획물 50 μg/ml을 24시간 동안 처리시 involucrin과 filaggrin단백질 발현을 각각122.8%, 105%로 유의하게 증가시킴을 확인하였다. Elisa assay를 통해 분석한 결과, ethyl acetate와 butanol 분획물은 대조군에 비해 hyaluronic acid의 생성을 각각17%, 11% 증가시켰으며, 이는 hyaluronic acid synthesis-1의 mRNA 발현 증가에 의한 것임을 확인하였다. 그러나 회향 열매 메탄올 추출물과 분획물 모두에서 β-defensin -1, -2, -3, LL-37의 mRNA 발현은 증가되지 않았다. 이러한 결과는 회향 열매 butanol 분획물이 각질형성세포에서 물리적 장벽을 형성하고 보습인자 조절을 통해 피부장벽 기능을 강화하는데 효과적임을 의미한다.

• Journal of Periodontal and Implant Science
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• 제24권3호
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• pp.472-482
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• 1994

Cytokines expressed specifically in leukocytes subsets and in activated cells, which are involved in chemotaxis and activation of leukocytes, are recently defined as chemokines. Macrophage inflammatory $protein-1{\alpha}(MIP-1{\alpha})$ and $MIP-1{\beta}$ are members of C-C chemokine subfamily which produces wide immunomodulatory, proinflammatory, and hematopoietic modulatory actions. We have studied their gene expression by using Northern blot analysis in various blood cells such as cytolytic T lymphocyte(CTL), helper T lymphocyte(HTL), macrophage, and B lymphocyte. Resting CTL line CTLL-R8 expressed $MIP-1{\alpha}$ mRNA which was downregulated by ConA stimulation. Both of resting and ConA stimulated HTL line Hut78 and Jurkat did not express $MIP-1{\alpha}$ mRNA. There was detectable $MIP-1{\alpha}$ transcript in HTL hybridoma 2B4.11 which was a little upstimulated by ConA stimulation. B cell line 230, and macrophage cell line RAW264.7 and WR19M.1 showed distinct $MIP-1{\alpha}$ message which were induced after LPS stimulation. Expression pattern of $MIP-1{\beta}$ in all cell lines or cell were almost identical to that of $MIP-1{\alpha}$. Also strategies employed to identify and characterize the biological functions was preceded by receptor cloning to trace the shorcut to the final goal of cytokine research. For the cloning of $MIP-1{\alpha}$ receptor(R), we used synthetic oligonucleotides of transmembrane(T) conserved sequences of already cloned human(h) IL-8-R, and performed reverse transcription-polymerase chain reaction(RT-PCR) amplification using murine(m) macrophage cell line mRNA. Among 5RT-PCR products, we isolated a homologous cDNA with hIL-8-R which were shown to be putative mIL-8-R cDNA.