• Title/Summary/Keyword: Gene Medicine

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Toxoplasma gondii B1 Gene Detection in Feces of Stray Cats around Seoul, Korea and Genotype Analysis of Two Laboratory-Passaged Isolates

  • Jung, Bong-Kwang;Lee, Sang-Eun;Lim, Hyemi;Cho, Jaeeun;Kim, Deok-Gyu;Song, Hyemi;Kim, Min-Jae;Shin, Eun-Hee;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.53 no.3
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    • pp.259-263
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    • 2015
  • The increasing prevalence of Toxoplasma gondii infection in the human population in the Republic of Korea (= Korea) is due to various reasons such as an increase in meat consumption. However, the importance of cats in transmitting T. gondii infection through oocysts to humans has seldom been assessed. A total of 300 fecal samples of stray cats captured around Seoul from June to August 2013 were examined for T. gondii B1 gene (indicating the presence of oocysts) using nested-PCR. Fourteen (4.7%) of 300 cats examined were positive for B1 gene. Female cats (7.5%) showed a higher prevalence than male cats (1.4%). Cats younger than 3 months (5.5%) showed a higher prevalence than cats (1.5%) older than 3 months. For laboratory passage of the positive samples, the fecal suspension (0.2 ml) of B1 gene positive cats was orally inoculated into experimental mice. Brain tissues of the mice were obtained after 40 days and examined for the presence of tissue cysts. Two isolates were successfully passaged (designated KNIH-1 and KNIH-2) and were molecularly analyzed using the SAG5D and SAG5E gene sequences. The SAG5D and SAG5E gene sequences showed high homologies with the ME49 strain (less virulent strain). The results indicated the importance of stray cats in transmitting T. gondii to humans in Korea, as revealed by detection of B1 gene in fecal samples. T. gondii isolates from cats were successfully passaged in the laboratory for the first time in Korea.

Multidrug-Resistant Providencia Isolates Carrying $bla_{PER-1},\;bla_{VIM-2}$, and armA

  • Lee, Hee-Woo;Kang, Hee-Young;Shin, Kyeong-Seob;Kim, Jung-Min
    • Journal of Microbiology
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    • v.45 no.3
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    • pp.272-274
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    • 2007
  • During May to July 2004, three strains of Providencia spp. with multidrug-resistance (MDR) were isolated from urinary specimen of three patients hospitalized with a same hospital room. By PCR analysis, all three strains have been found to carry both VIM-2 type $metallo-{\beta}-lactamase$ gene and PER-1 type extended spectrum ${\beta}-lactamase$ gene. One out of three strains carried additional resistance gene, armA, 16S rRNA methylase gene responsible for high level resistance to aminoglycosides. To our knowledge, this is the first report on the identification of Providencia spp. simultaneously carrying $bla_{VIM-2},\;bla_{PER-1}$, and armA genes.

A female patient with Xp21 gene deletion syndrome

  • Kim, Jungeun;Lee, Hyunjoo;Na, Ji-Hoon;Lee, Young-Mock
    • Journal of Genetic Medicine
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    • v.18 no.2
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    • pp.101-104
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    • 2021
  • Xp21 contiguous gene deletion syndrome is associated with complex glycerol kinase deficiency, congenital adrenal hypoplasia, Duchene muscular dystrophy, and intellectual disability. Xp21 gene deletion syndrome is X-linked recessive, so most symptomatic patients are male, and only a few female symptomatic patients have been reported. We report the first female Korean case of an Xp21 deletion. NGS data were analyzed for copy number variation, and the Xp21 deletion (chr X: 29301056-31838200) was confirmed using real-time PCR.

Effect of Schisandrae Chinensis Fructus on Keratinocyte Damage by UV Irradiation (오미자(五味子)가 자외선유발 피부각질세포 손상에 미치는 효과)

  • Park, Hyung-Hoon;Lee, Jang-Suk;Yun, Hyuk;Hwang, Gwi-Seo;Chong, Myong-Soo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.26 no.3
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    • pp.330-337
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    • 2012
  • This study was performed to assess the Effect of SCF(Schisandrae Chinensis Fructus) on Keratinocyte Damage by UV irradiation. The effect of SCF were determined in UV irradiated HaCaT. We measured LDH release and NO release from HaCaT to elucidate the effect of SCF. And iNOS, TNF-${\alpha}$, COX-2, Bax, Bcl-2, Bcl-xL, c-jun, c-fos gene expression were determined in HaCat using real time PCR method. The results are as follows. SCF inhibited LDH-release, NO production in UV irradiated HaCaT. SCF increased the gene expression Bax, Bcl-2 and Bcl-xL protein in UV irradiated HaCaT. SCF suppressed the gene expression TNF-${\alpha}$ in UV irradiated HaCaT. SCF suppressed the gene expression iNOS, c-fos, and c-jun in UV irradiated HaCaT. SCF not affected the suppression of the gene expression COX-2 in UV irradiated HaCaT. The study showed SCF inhibited the cell damage in UV irradiated HaCaT.

Survivin Gene Functions and Relationships between Expression and Prognosis in Patients with Nasopharyngeal Carcinoma

  • Cai, Jun-Hong;Fu, Sheng-Miao;Tu, Zhi-Hua;Deng, Li-Qun;Liang, Zhu;Chen, Xin-Ping;Gong, Xuan-Ju;Wan, Li-Hua
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.6
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    • pp.2341-2345
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    • 2015
  • This study aimed to investigate the relationship between prognosis and protein and mRNA expression of an apoptotic inhibitor gene, survivin, in patients with nasopharyngeal carcinoma. Furthermore, functions of the survivin gene in the CNE2 nasopharyngeal carcinoma cell line were assessed. Immunohistochemistry and in situ hybridization were used in detecting the survivin protein and mRNA in 44 nasopharyngeal carcinoma specimens, and 30 chronic nasopharyngitis samples as controls. Survivin gene expression in CNE2 cell line was suppressed with an shRNA (short hairpin RNA). The positive ratios of expression for survivin protein and mRNA in nasopharyngeal carcinoma were 79.5% and 75.0% respectively, obviously higher than in the control group (p<0.01), and there is very good consistency between the two methods. The mean survival time of patients with higher survivin protein or mRNA expression was shorter than in patients with lower levelsv(p<0.01). Proliferation of the CNE2 cell line was distinctly inhibited by the shRNA. The results indicate that overexpression of the survivin gene plays an important role in onset and development of nasopharyngeal carcinoma, and it may be helpful for prognostic appraisal.

Molecular Cloning, Identification and Characteristics of a Novel Isoform of Carbamyl Phosphate Synthetase I in Human Testis

  • Huo, Ran;Zhu, Hui;Lu, Li;Ying, Lanlan;Xu, Min;Xu, Zhiyang;Li, Jianmin;Zhou, Zuomin;Sha, Jiahao
    • BMB Reports
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    • v.38 no.1
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    • pp.28-33
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    • 2005
  • A gene coding a novel isoform of carbamyl phosphate synthetase I (CPS1) was cloned from a human testicular library. As shown by cDNA microarray hybridization, this gene was expressed at a higher level in human adult testes than in fetal testes. The full length of its cDNA was 3831 bp, with a 3149 bp open reading frame, encoding a 1050-amino-acid protein. The cDNA sequence was deposited in the GenBank (AY317138). Sequence analysis showed that it was homologous to the human CPS1 gene. The putative protein contained functional domains composing the intact large subunit of carbamoyl phosphate synthetase, thus indicated it has the capability of arginine biosynthesis. A multiple tissue expression profile showed high expression of this gene in human testis, suggesting the novel alternative splicing form of CPS1 may be correlated with human spermatogenesis.

Increases in Doxorubicin Sensitivity and Radioiodide Uptake by Transfecting shMDR and Sodium/Iodide Symporter Gene in Cancer Cells Expressing Multidrug Resistance (다약제내성 암세포에서 shMDR과 Sodium/Iodide Symporter 유전자의 이입에 의한 Doxorubicin 감수성과 방사성옥소 섭취의 증가)

  • Ahn, Sohn-Joo;Lee, Yong-Jin;Lee, You-La;Choi, Chang-Ik;Lee, Sang-Woo;Yoo, Jeong-Soo;Ahn, Byeong-Cheol;Lee, In-Kyu;Lee, Jae-Tae
    • Nuclear Medicine and Molecular Imaging
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    • v.41 no.3
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    • pp.209-217
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    • 2007
  • Purpose: Multidrug resistance (MDR) of the cancer cells related to mdr1 gene expression can be effectively treated by selective short hairpin RNA for mdr1 gene (shMDR). Sodium/iodide symporter (NIS) gene is well known to have both reporter and therapeutic gene characteristics. We have co-transfected both shMDR and NIS gene into colon cancer cells (HCT15 cell) expressing MDR and Tc-99m sestamibi and I-125 uptake were measured. In addition, cytotoxic effects of doxorubicin and I-131 therapy were also assessed after transfection. Material and Methods: At first, shMDR was transfected with liposome reagent into human embryonic kidney cells (HEK293) and HCT cells. shMDR transfection was confirmed by RT-PCR and western blot analysis. Adenovirus expressing NIS (Ad-NIS) gene and shMDR (Ad-shMDR) were co-transfected with Ad-NIS into HCT15 cells. Forty-eight hours after infection, inhibition of P-gycoprotein (Pgp) function by shMDR was analyzed by a change of Tc-99m sestamibi uptake and doxorubicin cytotoxicity, and functional activity of induced NIS gene expression was assessed with I-125 uptake assay. Results: In HEK293 cells transfected with shMDR, mdr1 mRNA and Pgp protein expressions were down regulated. HCT15 cells infected with 20 MOI of Ad-NIS was higher NIS protein expression than control cells. After transfection of 300 MOI of Ad-shMDR either with or without 10 MOI of Ad-NIS, uptake of Tc-99m sestamibi increased up to 1.5-fold than control cells. HCT15 cells infected with 10 MOI of Ad-NIS showed approximately 25-fold higher I-125 uptake than control cells. Cotransfection of Ad-shMDR and Ad-NIS resulted in enhanced cytotoxic by doxorubicin in HCT15 cells. I-131 treatment on HCT15 cells infected with 20 MOI of Ad-NIS revealed increased cytotoxic effect. Conclusion: Suppression of mdr1 gene expression, retention of Tc-99m sestamibi, enhanced doxorubicin cytotoxicity and increases in I-125 uptake were achieved in MDR expressing cancer cell by co-transfection of shMDR and NIS gene. Dual therapy with doxorubicin and radioiodine after cotransfection shMDR and NIS gene can be used to overcome MDR.

Derivation of Neural Precursor Cells from Human Embryonic Stem Cells

  • Kim Sehee;Hong Ji Young;Joo So Yeon;Kim Jae Hwan;Moon Shin Yong;Yoon Hyun Soo;Kim Doo Han;Chung Hyung Min;Choi Seong-Jun
    • Reproductive and Developmental Biology
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    • v.28 no.4
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    • pp.247-252
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    • 2004
  • Human embryonic stem (ES) cells are derived from the inner cell mass of the preimplantation embryo. Human ES cells have the capacity to differentiate into various types of cells in the body. Human ES cells are indefinite source of cells for cell therapy in various degenerative disorders including neuronal disorders. Directed differentiation of human ES cells is a prerequisite for their clinical application. The objective of this study is to develop the culture condition for the derivation of neural precursor cells from human ES cells. Neural precursor cells were derived from human ES cells in a stepwise culture condition. Neural precursor cells in the form of neural rosette structures developed into neurospheres when cultured in suspension. Suspension culture of neurospheres has been maintained over 4 months. Expressions of nestin, soxl, sox2, pax3 and pax6 transcripts were upregulated during differentiation into neural precursor cells by RT-PCR analysis. In contrast, expression of oct4 was dramatically downregulated in neural precursor cells. Immunocytochemical analyses of neural precursor cells demonstrated expression of nestin and SOX1. When induced to differentiate on an adhesive substrate, neuro-spheres were able to differentiate into three lineages of neural systems, including neurons, astrocytes and oligo-dendrocytes. Transcripts of sox1 and pax6 were downregulated during differentiation of neural precursor cells into neurons. In contrast, expression of map2ab was elevated in the differentiated cells, relative to those in neural precursor cells. Neurons derived from neural precursor cells expressed NCAM, Tuj1, MAP2ab, NeuN and NF200 in immunocytochemical analyses. Presence of astrocytes was confirmed by expression of GFAP immuno-cytochemically. Oligodendrocytes were also observed by positive immuno-reactivities against oligodendrocyte marker O1. Results of this study demonstrate that a stepwise culture condition is developed for the derivation of neural precursor cells from human ES cells.

Cervical Cancer Gene Therapy by Gene Loaded PEG-PLA Nanomedicine

  • Liu, Bo;Han, Shu-Mei;Tang, Xiao-Yong;Han, Li;Li, Chang-Zhong
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.12
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    • pp.4915-4918
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    • 2014
  • Background and Aims: Advances in the treatment of cervical cancer over the last decade have predominantly involved the development of genes directed at molecular targets. Gene therapy is recognized to be a novel method for the treatment of cervical cancer. Genes can be administered into target cells via nanocarriers. This study aimed to develop systemically administrable nano-vectors. Floate (Fa) containing gene loaded nanoparticles (NPs) could target HeLa human cervical cancer cells through combination with receptors on the cells to increase the nuclear uptake of genetic materials. Methods: Fa was linked onto Poly (ethylene glycol)-b-poly (D, L-lactide) (PEG-PLA) to form Fa-PEG-PLA, and the resulting material was used to load plasmids of enhanced green fluorescence protein (pEGFP) to obtain gene loaded nanoparticles (Fa-NPs/DNA). Physical-chemical characteristics, in vitro release and cytotoxicity of Fa-NPs/DNA were evaluated. The in vitro transfection efficiency of Fa-NPs/DNA was evaluated in HeLa cells and human umbilical vein endothelial cells (HUVEC). PEG-PLA without Fa was used to load pEGFP from NPs/DNA as a control. Results: Fa-NPs/DNA has a particle size of 183 nm and a gene loading quantity of 92%. After 72h of transfection, Fa-NPs/DNA displayed over 20% higher transfection efficiency than NPs/DNA and 40% higher than naked DNA in HeLa cells. However, in HUVECs, no significant difference appeared between Fa-NPs/DNA and NPs/DNA. Conclusions: Fa-PEG-PLA NPs could function as excellent materials for gene loading. This nano-approach could be used as tumor cell targeted medicine for the treatment of cervical cancer.

Magnolol Inhibits LPS-induced NF-${\kappa}B$/Rel Activation by Blocking p38 Kinase in Murine Macrophages

  • Li, Mei Hong;Kothandan, Gugan;Cho, Seung-Joo;Huong, Pham Thi Thu;Nan, Yong Hai;Lee, Kun-Yeong;Shin, Song-Yub;Yea, Sung-Su;Jeon, Young-Jin
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.6
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    • pp.353-358
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    • 2010
  • This study demonstrates the ability of magnolol, a hydroxylated biphenyl compound isolated from Magnolia officinalis, to inhibit LPS-induced expression of iNOS gene and activation of NF-${\kappa}B$/Rel in RAW 264.7 cells. Immunohisto-chemical staining of iNOS and Western blot analysis showed magnolol to inhibit iNOS gene expression. Reporter gene assay and electrophoretic mobility shift assay showed that magnolol inhibited NF-${\kappa}B$/Rel transcriptional activation and DNA binding, respectively. Since p38 is important in the regulation of iNOS gene expression, we investigated the possibility that magnolol to target p38 for its anti-inflammatory effects. A molecular modeling study proposed a binding position for magnolol that targets the ATP binding site of p38 kinase (3GC7). Direct interaction of magnolol and p38 was further confirmed by pull down assay using magnolol conjugated to Sepharose 4B beads. The specific p38 inhibitor SB203580 abrogated the LPS-induced NF-${\kappa}B$/Rel activation, whereas the selective MEK-1 inhibitor PD98059 did not affect the NF-${\kappa}B$/Rel. Collectively, the results of the series of experiments indicate that magnolol inhibits iNOS gene expression by blocking NF-${\kappa}B$/Rel and p38 kinase signaling.