• Asian Pacific Journal of Cancer Prevention
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• 제15권4호
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• pp.1675-1679
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• 2014

Objectives: MicroRNAs (miRNAs) are important regulators of many physiological and pathological processes, including tumorigenesis and metastasis. In this study, we sought to determine the underlying molecular mechanisms of metastatic cervical carcinoma by performing miRNA profiling. Methods: Tissue samples were collected from ten cervical squamous cancer patients who underwent hysterectomy and pelvic lymph node (PLN) dissection in our hospital, including four PLN-positive (metastatic) cases and six PLN-negative (non-metastatic) cases. A miRNA microarray platform with 1223 probes was used to determine the miRNA expression profiles of these two tissue types and case groups. MiRNAs having at least 4-fold differential expression between PLN-positive and PLN-negative cervical cancer tissues were bioinformatically analyzed for target gene prediction. MiRNAs with tumor-associated target genes were validated by quantitative reverse transcription-polymerase chain reaction (RT-PCR). Results: Thirty-nine miRNAs were differentially expressed (>4-fold) between the PLN-positive and PLN-negative groups, of which, 22 were up-regulated and 17 were down-regulated. Sixty-nine percent of the miRNAs (27/39) had tumor-associated target genes, and the expression levels of six of those (miR-126, miR-96, miR-144, miR-657, miR-490-5p, and miR-323-3p) were confirmed by quantitative (q)RT-PCR. Conclusions: Six MiRNAs with predicted tumor-associated target genes encoding proteins that are known to be involved in cell adhesion, cytoskeletal remodeling, cell proliferation, cell migration, and apoptosis were identified. These findings suggest that a panel of miRNAs may regulate multiple and various steps of the metastasis cascade by targeting metastasis-associated genes. Since these six miRNAs are predicted to target tumor-associated genes, it is likely that they contribute to the metastatic potential of cervical cancer and may aid in prognosis or molecular therapy.

• Asian-Australasian Journal of Animal Sciences
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• 제31권1호
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• pp.3-12
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• 2018

Objective: DNA methylation plays a major role in regulating the expression of genes related to traits of economic interest (e.g., weight gain) in livestock animals. This study characterized and investigated the functional inferences of genome-wide DNA methylome in the loin (longissimus dorsi) muscle (LDM) of swine. Methods: A total of 8.99 Gb methylated DNA immunoprecipitation sequence data were obtained from LDM samples of eight Duroc pigs (four pairs of littermates). The reference pig genome was annotated with 78.5% of the raw reads. A total of 33,506 putative methylated regions (PMR) were identified from methylated regions that overlapped at least two samples. Results: Of these, only 3.1% were commonly observed in all eight samples. DNA methylation patterns between two littermates were as diverse as between unrelated individuals (p = 0.47), indicating that maternal genetic effects have little influence on the variation in DNA methylation of porcine LDM. The highest density of PMR was observed on chromosome 10. A major proportion (47.7%) of PMR was present in the repeat regions, followed by introns (21.5%). The highest conservation of PMR was found in CpG islands (12.1%). These results show an important role for DNA methylation in species- and tissue-specific regulation of gene expression. PMR were also significantly related to muscular cell development, cell-cell communication, cellular integrity and transport, and nutrient metabolism. Conclusion: This study indicated the biased distribution and functional role of DNA methylation in gene expression of porcine LDM. DNA methylation was related to cell development, cell-cell communication, cellular integrity and transport, and nutrient metabolism (e.g., insulin signaling pathways). Nutritional and environmental management may have a significant impact on the variation in DNA methylation of porcine LDM.

• 한국동물학회지
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• 제38권4호
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• pp.505-513
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• 1995

P-lement의 germ line transformation을 이용하여 초파리 난소에서의 세포특이적 유전자발현 양상을 조사하였다. 표지유전자인 lacZ의 발현양상은 생식세포, 체세포, 두 종류의 세포에서 모두 발현되는 경우로 대별되며, 생식세포에서 lacZ의 발현은 조세포 또는 조세포와 난자에서 동시적으로 발현되는 경우로 구분되었다. 난자주변 난포세포에서의 lacZ 발현은 난자형성 9-10기 난포의 경우 모든 세포에서 균일하게 발현되거나 일부 세포에서만 발현되는 양상으로 구분되었다. 일부의 난포세포에서 발현되는 경우 난자와 조세포 경계의 난포세포에서 발현되는 경우, 이들을 제외한 난포세포에서 발현되는 경우로 구분되었다. 난자주위의 난포세포에서 난자의 전후축을 따른 구배현상도 관찰되었다. 극세포 또는 경계세포에서 발현되는 3개의 독립라인을 분리하였다. 독립된 EDL의 난소에서 나타난 lacZ의 발현양상은 초파리 난자형성에 관여하는 유전자의 시간과 공간적인 발현을 반영하며 이들 유전자 분리의 기초를 제공할 것으로 사료된다.

• Neonatal Medicine
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• 제28권3호
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• pp.133-138
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• 2021

Osteopetrosis refers to a group of genetic skeletal disorders characterized by osteosclerosis and fragile bones. Osteopetrosis can be classified into autosomal dominant, autosomal recessive, or X-linked forms, which might differ in clinical characteristics and disease severity. Autosomal recessive osteopetrosis, also known as malignant osteopetrosis, has an earlier onset, more serious clinical symptoms, and is usually fatal. We encountered a 1-day-old girl who was born full-term via vaginal delivery, which was complicated by meconium-stained amniotic fluid, cephalo-pelvic disproportion, and nuchal cord. Routine neonatal care was provided, in addition to blood tests and chest radiography to screen for sepsis, as well as skull radiography to rule out head injuries. Initial blood tests revealed hypocalcemia, which persisted on follow-up tests the next day. Radiographic examinations revealed diffusely increased bone density and a "space alien" appearance of the skull. Based on radiographic and laboratory findings, the infantile form of osteopetrosis was suspected and genetic testing for identification of the responsible gene. Eventually, a heterozygous mutation of the T cell immune regulator 1, ATPase H+ transporting V0 subunit a3 (TCIRG1) gene (c.292C>T) was identified, making this the first reported case of neonatal-onset malignant osteopetrosis with TCIRG1 mutation in South Korea. Early-onset hypocalcemia is common and usually results from prematurity, fetal growth restriction, maternal diabetes, perinatal asphyxia, and physiologic hypoparathyroidism. However, if hypocalcemia persists, we recommend considering 'infantile of osteopetrosis' as a rare cause of neonatal hypocalcemia and performing radiographic examinations to establish the diagnosis.

• Journal of Genetic Medicine
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• 제20권1호
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• pp.25-29
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• 2023

The CYP11A1 gene encodes for the cholesterol side-chain cleavage enzyme (P450scc), which initiates steroid hormone biosynthesis. Defective P450scc activity results in severe glucocorticoid and mineralocorticoid deficiencies. We describe a case of P450scc deficiency due to a novel homozygous CYP11A1 variant inherited from the mother with a possibility of uniparental disomy (UPD). The patient was a female, had no family history of endocrine disease, and showed adrenal insufficiency at 13 days of age. Hormonal analysis with an adrenocorticotropic hormone stimulation test showed both glucocorticoid and mineralocorticoid deficiencies, presumed to be a defect of the early stage of steroidogenesis. Exome sequencing reported a novel homozygous frameshift variant of CYP11A1 (c.284_285del, p.Asn95Serfs*10), which was inherited from the mother. Additionally, homozygosity in 15q22.31q26.2, which included CYP11A1, was identified using a chromosomal microarray. It was suggested that the possibility of maternal UPD was involved as the cause of a P450scc deficiency by unmasking the maternally derived affected allele. To our understanding, P450scc deficiency associated with UPD encompassing CYP11A1 had not been reported in Korea before. Genetic analysis can help diagnose rare causes of primary adrenal insufficiency, including P450scc deficiency.

• BMB Reports
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• 제36권2호
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• pp.179-184
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• 2003

Duchenne muscular dystrophy (DMD) is the most common hereditary neuromuscular disease. It is inherited manifestations. In some rare cases, the disease can also be manifested in females. The aim of the present study was to determine the molecular alteration in two cases of nonrelated DMD symptomatic carriers with no previous history of DMD. Multiplex PCR is commonly used to search for deletion in the DMD gene of affected males. This method could not be used in females because the normal X chromosome masks the deletion of the mutated one. Therefor, we used a set of seven highly polymorphic dinucleotide $(CA)_n$ repeat markers that lie within the human dystrophin gene. The deletions were evidenced by hemizygosity of the loci under study. We localized a deletion in the locus 7A (intron 7) on the maternal X chromosome in one case, and a deletion in the region of introns 49 and 50 on the paternal X chromosome in the other. The use of microsatellite genotyping within the DMD gene enables the detection of the mutant allele in female carriers. It is also a useful method to provide DMD families with more accurate genetic counseling.

• 대한바이러스학회지
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• 제26권1호
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• pp.9-22
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• 1996

Respiratory Syncytial virus (RSV) is an important cause of acute lower respiratory tract infections in human, with infants and young children being particularly susceptible. In the temperate zones, sharp annual outbreaks of RSV occur during the colder months, in both the northern and the southern hemisphere. RSV is unusual in that it can repeatedly reinfect individuals throughout life and infect babies in the presence of maternal antibody. RSV isolates can be divided into two subgroups, A and B, on the basis of their reactions with monoclonal antibodies, and the two subgroups are also distinct at the nucleotide sequence level. The specific diagnosis of RSV infection was best made by isolation of virus in tissue culture, identification of viral antigen, or by specific serologic procedures. Recently, rapid detection of RSV and analysis of RSV strain variation became possible by development of methods of reverse transcription and polymerase chain reaction amplification. In this study, to determine the genetic diversity of RSV found in Korea, 173 bp and 164 bp spanning selected regions of the RSV F and SH genes were enzymatically amplified and sequenced, respectively. Eight for F gene and three for SH gene were detected in 66 nasopharyngeal swap samples tested. Two major antigenic subgroups, A and B were confirmed from Korean samples (seven for subgroup A and one for subgroup B). At the nucleotide level of the F gene region, Korean subgroup A strains showed 95-99% homologies compared to the prototype A2 strain of subgroup A and 93-100% homologies among Korean subgroup A themselves. For the SH gene region, Korean subgroup A strain showed 97.5% homology compared to the prototype A2 strain of subgroup A, and Korean subgroup B strain showed 97% homology compared to the prototype 18537 strain of subgroup B. Most of base changes were transition and occured in codon position 3, which resulted in amino acid conservation. Using the maximum parsimony method, phylogenetic analysis indicated that Korean RSV strains formed a group with other RSV strains isolated from the United States, Canada, the Great Britain and Australia.

• 한국해양학회지:바다
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• 제5권2호
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• pp.157-168
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• 2000

우리나라 동해안에 서식하는 새치성게(Strongylocentrotus intermedius)는 둥근성게과(Strongylocentrotidae)에 속하는 냉수성 해양 무척추동물이다. 둥근성게과에는 현재 9종의 성게가 속해 있으나, 아직 종간의 분류 기준, 계통 분류학적 유연관계, 진화과정 등이 잘 밝혀져 있지 않다. 본 연구는 유전자 염기서열이라는 분자형질을 이용하여 새치성게의 종 분류기준을 확립하고 이 종의 계통진화 및 분화 시기를 파악하고자 수행되었다. 이를 위하여 변화율이 빠르고 모계로만 유전되는 특성을 가진 미토콘드리아의 한 유전자인 cytochrome c oxidase subunit I(COI)을 분석하였다. 새치성게의 생식소에서 DNA를 추출하고 중합효소연쇄반응으로 COI 유전자 단편을 선택적으로 증폭하였으며, 클로닝과 시퀀싱 과정을 거쳐 COI 유전자의 단편 1077개 염기쌍 순서(염기서열)를 확정하였다. 이 염기서열과 유전자 데이터베이스(GenBank)에 들어있는 다른 성게 및 해삼, 불가사리의 유전자를 비교하고 그 분자 계통수를 작성함으로써 새치성게의 진화과정을 분석하였다. COI 유전자 계통수는 새치성게가 태평양 동쪽 연안에 서식하는 S. purpuratus와 계통적으로 자매군(sister species)의 관계에 있음을 보였다. 두 종의 분화 시기는 계통수 상 분지의 길이와 화석연대를 고려하여 산출했을 때 지구 온도의 변동이 심했던 약 890만년 전으로 추정되었다. 태평양의 동안과 서안으로 분리된 두 종의 현재 분포와 종분화 시기의 지구 환경조건은 두 종간의 분화가 환경변화에 따른 개체군의 지리적 분리(vicariance)에 의한 것임을 시사해 준다. 한편, 새치성게의 COI 유전자염기서열은 이 종을 대표하는 분자형질로서 둥근성게과의 성게들을 서로 구분할 수 있는 종분류의 기준이 될 것이다.

• Asian-Australasian Journal of Animal Sciences
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• 제20권2호
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• pp.178-183
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• 2007

China has numerous native domestic goat breeds, but so far there has been no extensive study on genetic diversity, population demographic history, and origin of Chinese goats. To determine the origin and genetic diversity of Chinese goats, we analyzed the complete mtDNA D-loop sequences of 183 goats from 13 breeds. The haplotype diversity value found in each breed ranged from 0.9333 to 1.0000. The nucleotide diversity value ranged from 0.006337 to 0.025194. Our results showed that there were four mtDNA lineages (A, B, C and D), in which lineage A was predominant, lineage B was moderate, and lineages C and D were at low frequencies. Lineages C and D were observed only in the Tibetan breed. The results revealed multiple maternal origins of Chinese domestic goats. There was weaker geographical structuring in the 13 Chinese goat populations, which suggested that there existed high gene flow among goat populations caused by the extensive transportation of goats in the course of history.

• Clinical and Experimental Reproductive Medicine
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• 제40권1호
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• pp.12-22
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• 2013

Objective: We investigated the norepinephrine transporter (NET) expression in normal and pre-eclamptic placentas and analyzed the invasion activity of trophoblastic cells based on norepinephrine (NE)-NET regulation. Methods: NET and NE expression levels were examined by western blot and enzyme-linked immunosorbent assay, respectively. Trophoblast invasion activity, depending on NE-NET regulation, was determined by NET-small interfering RNA (siRNA) and NET transfection into the human extravillous trophoblast cells with or without NE treatment and invasion rates were analyzed by zymography and an invasion assay. Results: NET mRNA was expressed at a low level in pre-eclamptic placentas compared with normal placentas and NE concentration in maternal plasma increased significantly in pre-eclamptic women compared to normal pregnant women (p<0.05). NET gene upregulation and NE treatment stimulated trophoblast cell invasion up to 2.5-fold (p<0.05) by stimulating matrix metalloproteinase-9 activity via the phosphoinositol-3-kinase/AKT signaling pathway, whereas NET-siRNA with NE treatment reduced invasion rates. Conclusion: NET expression is reduced by inadequate regulation of NE levels during placental development. This suggests that a complementary balance between NET and NE regulates trophoblast cell invasion activities during placental development.