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Long Non-coding RNAs are Differentially Expressed in Hepatocellular Carcinoma Cell Lines with Differing Metastatic Potential

  • Fang, Ting-Ting;Sun, Xiao-Jing;Chen, Jie;Zhao, Yan;Sun, Rui-Xia;Ren, Ning;Liu, Bin-Bin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10513-10524
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    • 2015
  • Background: Metastasis is a major reason for poor prognosis in patients with cancer, including hepatocellular carcinoma (HCC). A salient feature is the ability of cancer cells to colonize different organs. Long non-coding RNAs (lncRNAs) play important roles in numerous cellular processes, including metastasis. Materials and Methods: In this study, the lncRNA expression profiles of two HCC cell lines, one with high potential for metastasis to the lung (HCCLM3) and the other to lymph nodes (HCCLYM-H2) were assessed using the Arraystar Human LncRNA Array v2.0, which contains 33,045 lncRNAs and 30,215 mRNAs. Coding-non-coding gene co-expression (CNC) networks were constructed and gene set enrichment analysis (GSEA) was performed to identify lncRNAs with potential functions in organ-specific metastasis. Levels of two representative lncRNAs and one representative mRNA, RP5-1014O16.1, lincRNA-TSPAN8 and TSPAN8, were further detected in HCC cell lines with differing metastasis potential by qRT-PCR. Results: Using microarray data, we identified 1,482 lncRNAs and 1,629 mRNAs that were differentially expressed (${\geq}1.5$ fold-change) between the two HCC cell lines. The most upregulated lncRNAs in H2 were RP11-672F9.1, RP5-1014O16.1, and RP11-501G6.1, while the most downregulated ones were lincRNA-TSPAN8, lincRNA-CALCA, C14orf132, NCRNA00173, and CR613944. The most upregulated mRNAs in H2 were C15orf48, PSG2, and PSG8, while the most downregulated ones were CALCB, CD81, CD24, TSPAN8, and SOST. Among them, lincRNA-TSPAN8 and TSPAN8 were found highly expressed in high lung metastatic potential HCC cells, while lowly expressed in no or low lung metastatic potential HCC cells. RP5-1014O16.1 was highly expressed in high lymphatic metastatic potential HCC cell lines, while lowly expressed in no lymphatic metastatic potential HCC cell lines. Conclusions: We provide the first detailed description of lncRNA expression profiles related to organ-specific metastasis in HCC. We demonstrated that a large number of lncRNAs may play important roles in driving HCC cells to metastasize to different sites; these lncRNAs may provide novel molecular biomarkers and offer a new basis for combating metastasis in HCC cases.

Production of Recombinant Human Hyperglycosylated Erythropoietin Using Cell Culture Technology by Improving Sialylation. (Sialic Acid 함량 증가 배양기술에 의한 재조합 인간 다당쇄 에리스로포이에틴의 생산)

  • 박세철;이승오;박만식;김승훈;김준환;송무영;이병규;고인영;강희일
    • Microbiology and Biotechnology Letters
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    • v.32 no.2
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    • pp.142-148
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    • 2004
  • Erythropoietin is a main regulator of human erythropoiesis. Recombinant human erythropoietin (rhEPO) is one of the glycoproteins produced in animal cells, and it has oligo saccharides chains which comprise about 40% of its molecular mass. Because the content of sialic acid can extend circulatory lifetime, the high degree of sialylation is often a desirable feature of therapeutic glycoproteins. In this study, the sialylation of rhEPO produced by chinese hamster ovary cell culture was maximized by supplementing the culture medium with N-acetylm-annosamine (ManNAc), a direct intracellular precursor for sialic acid synthesis and 2-deoxy-2,3-dehydro-N-acetylneuraminic acid (NeuAc2en), a sialidase inhibitor. Feeding of 20 mM ManNAc/0.5 mM NeuAc2en into culture medium increased the sialic acid content by nearly tenfold compared with unsupplemented medium. This effect was achieved without affecting the cell growth or product yield. Six erythropoietin fractions differing in sialic acid content, ranging from 11∼15% of EPO, were identified from chinese hamster ovary cell-derived rhEPO by mono Q column chromatography. It was found that, at 20 mM ManNAc/0.5 mM NeuAc2en feeding, productivity of hyper-glycosylated EPO increased up to 50%, compared with the unsupplemented medium.

Early Diagnosis of KBG Syndrome Using Diagnostic Exome Sequencing (Diagnostic exome sequencing을 통한 KBG 증후군의 조기 진단)

  • Hong, Jun Ho;Kim, Se Hee;Lee, Seung Tae;Choi, Jong Rak;Kang, Hoon Chul;Lee, Joon Soo;Kim, Heung Dong
    • Journal of the Korean Child Neurology Society
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    • v.26 no.4
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    • pp.272-275
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    • 2018
  • KBG syndrome is a rare neurodevelopmental disorder characterized by intellectual disability, skeletal anomalies, short stature, craniofacial dysmorphism, and macrodontia. ANKRD11 gene mutation and 16q24.3 microdeletion have been reported to cause KBG syndrome. Here, we report two patients with ANKRD11 mutations who initially presented with neurologic symptoms such as developmental delay and seizures. Patient 1 was a 23-month-old boy who presented with a global developmental delay. Language delay was the most dominant feature. He had hypertelorism, hearing impairment, and behavior problems characterized as hyperactivity. A c.1903_1907delAAACA (p.Lys635GInfsTer26) mutation in ANKRD11 was identified with diagnostic exome sequencing. Patient 2 was a 14-month-old boy with developmental delay and seizure. He also had atrial septum defect, and ventricular septal defect. Generalized tonic seizures began at the age of 8 months. Electroencephalography showed generalized sharp and slow wave pattern. Seizures did not respond to antiepileptic drugs. A loss of function mutation c.5350_5351delTC (p.ser1784HisfsTer12) in ANKRD11 was identified with diagnostic exome sequencing. In both cases, characteristic features of KBG syndrome such as short stature or macrodontia, were absent, and they visited the hospital due to neurological symptoms. These findings suggest that more patients with mild phenotypes of KBG syndrome are being recognized with advances in diagnostic exome sequencing genetic technologies.

Integrated Photonic Microwave Bandpass Filter Incorporating a Polymer Microring Resonator (폴리머 마이크로링 공진기를 이용한 광학적 마이크로웨이브 대역통과 필터)

  • Chin, Won-Jun;Kim, Do-Hwan;Song, Ju-Han;Lee, Sang-Shin
    • Korean Journal of Optics and Photonics
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    • v.16 no.5
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    • pp.469-475
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    • 2005
  • An integrated photonic microwave bandpass filter has been proposed and demonstrated incorporating a coherently coupled microring resonator in low-loss polymers. The proposed device may feature compact site, simple structure, tuning via the thermooptic and electrooptic effect, and flexible integration with other electrical and optical devices. The resonator was designed to have an extremely small bandwidth so that it could be used to selectively pass the optical signal carrying the microwave signal to attain efficient bandpass filtering. We made and tested two resonators with a single ring and double rings, and performed a theoretical fitting of their measured transfer curves to predict the performance of the microwave filters based on them precisely. It was found that as the number of the rings used for the resonator increases, the bandwidth gets smaller, the rolloff sharper, and the band rejection higher. Finally our filter exhibited the center frequency of 10GHz, the 3-dB bandwidth of 1.0GHz, the corresponding quality (Q) factor of 10, and the rejection outside of the passband of more than 25dB.

Effect of a Hot Water Extract of Sparasis Crispa on the Expression of Tight Junction-Associated Genes in HaCaT Cells (꽃송이버섯 열수추출물이 HaCaT의 세포 연접 관련 유전자의 발현에 대한 영향)

  • Han, Hyo-Sang
    • Journal of The Korean Society of Integrative Medicine
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    • v.9 no.2
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    • pp.83-92
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    • 2021
  • Purpose : Keratinocytes are the main cellular components involved in wound healing during re-epithelization and inflammation. Dysfunction of tight junction (TJ) adhesions is a major feature in the pathogenesis of various diseases. The purpose of this study was to identify the various effects of a Sparassis crispa water extract (SC) on HaCaT cells and to investigate whether these effects might be applicable to human skin. Methods : We investigated the effectiveness of SC on cell HaCaT viability using MTS. The antioxidant effect of SC was analyzed by comparing the effectiveness of ABTS to that of the well-known antioxidant resveratrol. Reverse-transcription quantitative polymerase chain reaction (qRT-PCR) is the most widely applied method Quantitative RT-PCR analysis has shown that SC in HaCaT cells affects mRNA expression of tight-junction genes associated with skin moisturization. In addition, Wound healing is one of the most complex processes in the human body. It involves the spatial and temporal synchronization of a variety of cell types with distinct roles in the phases of hemostasis, inflammation, growth, re-epithelialization, and remodeling. wound healing analysis demonstrated altered cell migration in SC-treated HaCaT cells. Results : MTS analysis in HaCaT cells was found to be more cytotoxic in SC at a concentration of 0.5 mg/㎖. Compared to 100 µM resveratrol, 4 mg/㎖ SC exhibited similar or superior antioxidant effects. SC treatment in HaCaT cells reduced levels of claudin 1, claudin 3, claudin 4, claudin 6, claudin 7, claudin 8, ZO-1, ZO-2, JAM-A, occludin, and Tricellulin mRNA expression by about 1.13 times. Wound healing analysis demonstrated altered cell migration in SC-treated HaCaT cells and HaCaT cell migration was also reduced to 73.2 % by SC treatment. Conclusion : SC, which acts as an antioxidant, reduces oxidative stress and prevents aging of the skin. Further research is needed to address the effects of SC on human skin given the observed alteration of mRNA expression of tight-junction genes and the decreased the cell migration of HaCaT cells.

Sphingomonas abietis sp. nov., an Endophytic Bacterium Isolated from Korean Fir

  • Lingmin Jiang;Hanna Choe;Yuxin Peng;Doeun Jeon;Donghyun Cho;Yue Jiang;Ju Huck Lee;Cha Young Kim;Jiyoung Lee
    • Journal of Microbiology and Biotechnology
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    • v.33 no.10
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    • pp.1292-1298
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    • 2023
  • PAMB 00755T, a bacterial strain, was isolated from Korean fir leaves. The strain exhibits yellow colonies and consists of Gram-negative, non-motile, short rods or ovoid-shaped cells. It displays optimal growth conditions at 20℃, 0% NaCl, and pH 6.0. Results of 16S rRNA gene-based phylogenetic analyses showed that strain PAMB 00755T was most closely related to Sphingomonas chungangi MAH-6T (97.7%) and Sphingomonas polyaromaticivorans B2-7T (97.4%), and ≤96.5% sequence similarity to other members of the genus Sphingomonas. The values of average nucleotide identity (79.9-81.3%), average amino acid identity (73.3-75.9%), and digital DNA-DNA hybridization (73.3-75.9%) were significantly lower than the threshold values for species boundaries; these overall genome-related indexes (OGRI) analyses indicated that the strain represents a novel species. Genomic analysis revealed that the strain has a 4.4-Mbp genome encoding 4,083 functional genes, while the DNA G+C content of the whole genome is 66.1%. The genome of strain PAMB 00755T showed a putative carotenoid biosynthetic cluster responsible for its antioxidant activity. The respiratory quinone was identified as ubiquinone 10 (Q-10), while the major fatty acids in the profile were identified as C18:1ω7c and/or C18:1ω6c (summed feature 8). The major polar lipids of strain PAMB 00755T were diphosphatidylglycerol, phosphatidylethanolamine, sphingoglycolipid, and phosphatidylcholine. Based on a comprehensive analysis of genomic, phenotypic, and chemotaxonomic characteristics, we proposed the name Sphingomonas abietis sp. nov. for this novel species, with PAMB 00755T as the type strain (= KCTC 92781T = GDMCC 1.3779T).

Understanding the protox inhibition activity of novel 1-(5-methyl-3-phenylisoxazolin-5-yl)methoxy-2-chloro-4-fluorobenzene derivatives using comparative molecular field analysis (CoMFA) methodology (비교 분자장 분석 (CoMFA) 방법에 따른 1-(5-methyl-3-phenylisoxazolin-5-yl)methoxy-2-chloro-4-fluoro-benzene 유도체들의 Protox 저해 활성에 관한 이해)

  • Sung, Nack-Do;Song, Jong-Hwan;Yang, Sook-Young;Park, Kyeng-Yong
    • The Korean Journal of Pesticide Science
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    • v.8 no.3
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    • pp.151-161
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    • 2004
  • Three dimensional quantitative structure-activity relationships (3D-QSAR) studies for the protox inhibition activities against root and shoot of rice plant (Orysa sativa L.) and barnyardgrass (Echinochloa crus-galli) by a series of new A=3,4,5,6-tetrahydrophthalimino, B=3-chloro-4,5,6,7-tetrahydro-2H-indazolyl and C=3,4-dimethylmaleimino group, and R-group substituted on the phenyl ring in 1-(5-methyl-3-phenylisoxazolin-5-yl)methoxy-2chloro-4-fluorobenzene derivatives were performed using comparative molecular field analyses (CoMFA) methodology with Gasteiger-Huckel charge. Four CoMFA models for the protox inhibition activities against root and shoot of the two plants were generated using 46 molecules as training set and the predictive ability of the each models was evaluated against a test set of 8 molecules. And the statistical results of these models with combination (SIH) of standard field, indicator field and H-bond field showed the best predictability of the protox inhibition activities based on the cross-validated value $r^2_{cv.}$ $(q^2=0.635\sim0.924)$, conventional coefficient $(r^2_{ncv.}=0.928\sim0.977)$ and PRESS value $(0.091\sim0.156)$, respectively. The activities exhibited a strong correlation with steric $(74.3\sim87.4%)$, electrostatic $(10.10\sim18.5%)$ and hydrophobic $(1.10\sim8.30%)$ factors of the molecules. The steric feature of molecule may be an important factor for the activities. We founded that an novel selective and higher protox inhibitors between the two plants may be designed by modification of X-subsitutents for barnyardgrass based upon the results obtained from CoMFA analyses.

Comparison of Tension Type Headache Associated with Pericranial Tenderness and Headache Attributed to Temporomandibular Joint Disorder Using RDC/TMD Axis II (두개주변압통과 관련된 긴장성 두통과 측두하악장애에 기인한 두통과의 RDC/TMD Axis II에 따른 비교)

  • Park, Hyung-Yun;Bae, Sung-Jae;Yoo, Sang-Hoon;Chun, Yang-Hyun;Hong, Jung-Pyo;Auh, Q-Schick
    • Journal of Oral Medicine and Pain
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    • v.35 no.2
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    • pp.123-133
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    • 2010
  • The purpose of this study is to investigate Tension-type headache associated with pericranial tenderness and Headache attributed to temporomandibular joint disorder among Characteristic pain index, Disability points, Chronic pain grade, Depression index, Nonspecific physical symptoms (pain items included), Nonspecific physical symptoms (pain items excluded), Jaw disability score using RDC/TMD Axis II. Eighty five patients who visited the Kyung Hee University Dental Hospital were diagnosed as Tension-type headache associated with pericranial tenderness(n=48) and Headache attributed to temporomandibular joint disorder(n=37) by the International Classification of Headache Disorders, 2nd Edition, and were administered the Korean versions of the RDC/TMD Axis II- Characteristic pain index, Disability points, Chronic pain grade, Depression index, Nonspecific physical symptoms (pain items included), Nonspecific physical symptoms (pain items excluded), Jaw disability score. Then it was analyzed statistically by SPSS(ver. 10.0). T-test, The Wilcoxon-signed rank test and Mann-Whitney U test (p<0.05) were used. There were no significant differences in Characteristic pain index, Disability points, Chronic pain grade, Depression index, Nonspecific physical symptoms (pain items included), Nonspecific physical symptoms (pain items excluded), Jaw disability score between two groups. Tension type headache associated with pericranial tenderness and Headache attributed to temporomandibular joint disorder showed similar Axis II feature. Tension-type headache associated with pericranial tenderness may be related to Headache attributed to temporomandibular joint disorder.

Applicability of plate tectonics to the post-late Cretaceous igneous activities and mineralization in the southern part of South Korea( I ) (한국남부(韓國南部)의 백악기말(白堊紀末) 이후(以後)의 화성활동(火成活動)과 광화작용(鑛化作用)에 대(對)한 판구조론(板構造論)의 적용성(適用性) 연구(硏究)( I ))

  • Min, Kyung Duck;Kim, Ok Joon;Yun, Suckew;Lee, Dai Sung;Joo, Sung Whan
    • Economic and Environmental Geology
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    • v.15 no.3
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    • pp.123-154
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    • 1982
  • Petrochemical, K-Ar dating, Sand Rb/Sr isotopes, metallogenic zoning, paleomagnetic and geotectonic studies of the Gyongsang basin were carried out to examine applicability of plate tectonics to the post-late Cretaceous igneous activity and metallogeny in the southeastern part of Korean Peninsula. The results obtained are as follows: 1. Bulgugsa granitic rocks range from granite to adamellite, whose Q-Ab-Or triangular diagram indicates that the depth and pressure at which the magma consolidated increase from coast to inland varying from 6 km, 0.5-3.3 kb in the coastal area to 17 km, 0.5-10 kb in the inland area. 2. The volcanic rocks in Gyongsang basin range from andesitic to basaltic rocks, and the basaltic rocks are generally tholeiitic in the coastal area and alkali basalt in the inland area. 3. The volcanic rocks of the area have the initial ratio of Sr^{87}/Sr^{86} varying from 0.706 to 0.707 which suggests a continental origin; the ratio of Rb/Sr changing from 0.079-0.157 in the coastal area to 0.021-0.034 in the inland area suggests that the volcanism is getting younger toward coastal side, which may indicate a retreat in stage of differentiation if they were derived from a same magma. The K_2O/SiO_2 (60%) increases from about 1.0 in the coastal area to about 3.0 in the inland area, which may suggest an increase indepth of the Benioff zone, if existed, toward inland side. 4. The K-Ar ages of volcanic rocks were measured to be 79.4 m.y. near Daegu, and 61.7 m.y. near Busan indicating a southeastward decrease in age. The ages of plutonic rocks also decrease toward the same direction with 73 m.y. near Daegu, and 58 m.y. near Busan, so that the volcanism predated the plutonism by 6 m.y. in the continental interior and 4 m.y. along the coast. Such igneous activities provide a positive evidence for an applicability of plate tectonics to this area. 5. Sulfur isotope analyses of sulfide minerals from 8 mines revealed that these deposits were genetically connected with the spacially associated ingeous rocks showing relatively narrow range of ${\delta}^{34}S$ values (-0.9‰ to +7.5‰ except for +13.3 from Mulgum Mine). A sequence of metallogenic zones from the coast to the inland is delineated to be in the order of Fe-Cu zone, Cu-Pb-Zn zone, and W-Mo zone. A few porphyry type copper deposits are found in the Fe-Cu zone. These two facts enable the sequence to be comparable with that of Andean type in South America. 6. The VGP's of Cretaceous and post Cretaceous rocks from Korea are located near the ones($71^{\circ}N$, $180^{\circ}E$ and $90^{\circ}N$, $110^{\circ}E$) obtained from continents of northern hemisphere. This suggests that the Korean peninsula has been stable tectonically since Cretaceous, belonging to the Eurasian continent. 7. Different polar wandering path between Korean peninsula and Japanese islands delineates that there has been some relative movement between them. 8. The variational feature of declination of NRM toward northwestern inland side from southeastern extremity of Korean peninsula suggests that the age of rocks becomes older toward inland side. 9. The geological structure(mainly faults) and trends of lineaments interpreted from the Landsat imagery reveal that NNE-, NWW- and NEE-trends are predominant in the decreasing order of intensity. 10. The NNE-trending structures were originated by tensional and/or compressional forces, the directions of which were parallel and perpendicular respectively to the subduction boundary of the Kula plate during about 90 m.y. B.P. The NWW-trending structures were originated as shear fractures by the same compressional forces. The NEE-trending structures are considered to be priginated as tension fractures parallel to the subduction boundary of the Kula plate during about 70 m.y. B.P. when Japanese islands had drifted toward southeast leaving the Sea of Japan behind. It was clearly demonstrated by many authors that the drifting of Japanese islands was accompanied with a rotational movement of a clock-wise direction, so that it is inferred that subduction boundary had changed from NNE- to NEE-direction. A number of facts and features mentioned above provide a suite of positive evidences enabling application of plate tectonics to the late Cretaceous-early Tertiary igneous activity and metallogeny in the area. Synthesizing these facts, an arc-trench system of continental margin-type is adopted by reconstructing paleogeographic models for the evolution of Korean peninsula and Japan islands. The models involve an extention mechanism behind the are(proto-Japan), by which proto-Japan as of northeastern continuation of Gyongsang zone has been drifted rotationally toward southeast. The zone of igneous activity has also been migrated from the inland in late-Cretaceous to the peninsula margin and southwestern Japan in Tertiary.

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