• Title/Summary/Keyword: IS-PCR

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Analysis of the factors influencing headache and backache following lumbar puncture (요추 천자 후 발생하는 두통 및 요통에 영향을 주는 요인의 분석)

  • Lee, Sang Taek;Chung, Sochung;Park, Yong Mean;Bae, Sun Hwan;Yu, Jeong Jin;Lee, Ran
    • Clinical and Experimental Pediatrics
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    • v.51 no.8
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    • pp.856-860
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    • 2008
  • Purpose : This study aimed to examine the factors influencing the appearance of headache and backache following diagnostic lumbar puncture in children, focusing on the need for strict bed rest after lumbar puncture. Methods : We studied 70 two-fifteen-year-old pediatric patients who underwent diagnostic lumbar puncture from July 2005 to July 2007 at Konkuk University Hospital. We divided them into two groups. Patients in the first group (n=24) were allowed free mobility and patients in the second group (n=46) were to have strict bed rest for four hours after puncture. Data were analyzed by age, sex, number of puncture attempts, cell counts and pressure in the cerebrospinal fluid (CSF), duration of bed rest, and occurrence of headache and backache. Results : The rate of complications was not significantly related to sex, age, presence of enterovirus, CSF pressure, or postural headache. The occurrence of headache was significantly correlated with white blood cell (WBC) count in CSF (P=0.043). Symptom frequency did not differ significantly between the groups. Backache was significantly related to the frequency of puncture attempts (P=0.046). Conclusion : Strict bed rest following diagnostic lumbar puncture in children does not influence headaches and backaches. These are respectively related to the WBC count on the CSF profile and the frequency of attempts. Therefore, after lumbar puncture, absolute bed rest is not necessary and patients are more comfortable with free mobility.

Single Nucleotide Polymorphisms of the GnRHR Gene Associated with Reproductive Traits of Japanese Flounder (Paralichthys olivaceus)

  • He, Feng;Wen, Hai-Shen;Li, Ji-Fang;Yu, Da-Hui;Ma, Rui-Qin;Shi, Dan;Mu, Wei-Jie;Zhang, Yuan-Qing;Hu, Jian;Liu, Miao;Han, Wei-Guo;Zhang, Jia-Nan;Wang, Qing-Qing;Yuan, Yu-Ren;Liu, Qun
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.4
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    • pp.463-470
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    • 2011
  • Gonadotropin-releasing hormone receptor (GnRHR) gene is expressed at the anterior pituitary gland and plays a key role in gonad development. This study aimed to investigate molecular genetic characteristics of the GnRHR gene and elucidate the effects of single nucleotide polymorphisms (SNPs) of GnRHR gene on sex steroid level in Japanese flounder (Paralichthys olivaceus). We used polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) and sequencing of the GnRHR gene in 75 individuals. We identified three SNPs in the GnRHR gene: P1 locus (C759A and C830T) in the coding region of exon2 which were both linked together and P2 locus (G984T) in the coding region of exon3, which added a new transcript factor (ADR1) and a new methylation site (CG). Only C830T of P1 leads to amino acid changes Thr266Ile. Statistical analysis showed that P1 was significantly associated with $17{\beta}$-estradiol ($E_2$) level (p<0.01) and gonadosomatic index (GSI) (p<0.05). Individuals with genotype BB of P1 had significantly higher serum $E_2$ levels (p<0.01) and GSI (p<0.05) than those of genotype AA or AB. Another SNP, P2, synonymous mutation, was significantly associated with GSI (p<0.05). Individuals with genotype AB of P2 had significantly higher GSI (p<0.05) than that of genotype AA. In addition, there was a significant association between one diplotype based on three SNPs and reproductive traits. The genetic effects for both serum $E_2$ level and GSI of diplotype D4 were super diplotypes (p<0.05). These results suggest that the SNPs in Japanese Flounder GnRHR are associated with $E_2$ level and GSI.

Association between Laryngeal Squamous Cell Carcinoma and Polymorphisms in Tumor Necrosis Factor Related Apoptosis Induce Ligand (TRAIL), TRAIL Receptor and sTRAIL Levels

  • Verim, Aysegul;Turan, Saime;Farooqi, Ammad Ahmad;Kahraman, Ozlem Timirci;Tepe-Karaca, Cigdem;Yildiz, Yemliha;Naiboglu, Baris;Ozkan, Nazli Ezgi;Ergen, Arzu;Isitmangil, Gulbu Aydinoglu;Yaylim, Ilhan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.24
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    • pp.10697-10703
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    • 2015
  • The laryngeal squamous cell carcinoma (LSCC) is one of the most common malignant tumors occurring in the head and neck. Tumor necrosis factor related apoptosis induce ligand (TRAIL) and TRAIL-receptors (DR4, DR5, DcR1, DcR2) are known as important members of TRAIL-mediated biochemical signaling pathway. Associations between polymorphisms in these genes and clinicopathological characteristics of human laryngeal carcinoma are not well defined. This study therefore aimed to investigate a possible relationship among the TRAIL and TRAIL-DR4 polymorphisms and sTRAIL levels in the risk or progression of LSCC. A total of 99 patients with laryngeal cancer and 120 healthy subjects were enrolled in the study. DR4 C626G and TRAIL 1595 C/T genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis and sTRAIL levels were measured by ELISA. There were significant differences in the distribution of DR4 C626G genotypes and frequencies of the alleles between laryngeal cancer patients and controls (p<0.001) but not in TRAIL 1595 C/T. We found the increased frequency of the DR4 C626G homozygote CC genotype in patients than in controls (p<0.001). Haplotype analysis revealed that there was also a statistically significant relationship between TRAIL and TRAIL-DR4 polymorphisms and laryngeal cancer. Serum sTRAIL levels in the laryngeal patients with CC genotype who had advanced tumour stage were lower than those of patients with early tumor stage (p=0.014). Our findings suggest that DR4 C626G genotypes and sTRAIL levels might be associated with progression of laryngeal cancer in the Turkish population.

Neuroprotective effects of herbal mixture HT070 on global cerebral ischemia in rats

  • Song, Jungbin;Lee, Donghun;Kim, Young-Sik;Lee, Hyun Jeong;Lee, Seunggyeong;Kim, Dong Kuk;Kang, Shin Ho;Shin, Yong Kook;Choi, Ho-Young;Kim, Hocheol
    • The Korea Journal of Herbology
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    • v.31 no.4
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    • pp.101-109
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    • 2016
  • Objectives : HT070 is a mixture of herbal extracts from root of Scutellaria baicalensis and stem bark of Eleutherococcus senticosus , which have long been used for stroke therapy in traditional Korean Medicine. The purpose of this study was to investigate the neuroprotective effects of HT070 on global cerebral ischemia and its potential mechanisms.Methods : Transient global cerebral ischemia was produced by 10 min of four-vessel occlusion (4-VO) in male Wistar rats. HT070 was administered orally at a dosage of 200 mg/kg twice at 0 and 90 min after reperfusion. Hippocampal neuronal damage was measured 7 days after reperfusion. To explore the potential mechanisms, we used hydrogen peroxide (H2O2)-induced rat pheochromocytoma (PC12) cells as an in vitro model. PC12 cells were pretreated with HT070 for 1 h and then exposed to 100 μM H2O2 for 6 h in the presence of HT070. Cell viability was measured by MTT assay and the mRNA expression of Bax, Bcl-2, iNOS and COX-2 were measured by quantitative RT-PCR.Results : Oral administration of HT070 at a dose of 200 mg/kg significantly reduced neuronal death in the hippocampal CA1 region by 13.4% as compared to the vehicle-treated group. HT070 increased cell viability, reversed the down-regulated Bcl-2 mRNA level, and suppressed the up-regulated mRNA expressions of Bax, iNOS, and COX-2 in H2O2-treated PC12 cells.Conclusions : HT070 protects against delayed neuronal death after global cerebral ischemia and its neuroprotection properties might be attributed to the inhibition of mitochondrial apoptosis and ROS-generating enzymes.

Comparing the anti-inflammatory effect of nanoencapsulated lycopene and lycopene on RAW 264.7 macrophage cell line (RAW 264.7 대식세포주에서 나노입자화 리코펜의 항염증 증진 효과)

  • Seo, Eun Young;Kim, Myung Hwan;Kim, Woo-Kyoung;Chang, Moon-Jeong
    • Journal of Nutrition and Health
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    • v.48 no.6
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    • pp.459-467
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    • 2015
  • Purpose: We developed a method to load lycopene into maltodextrin and cyclodextrin in an attempt to overcome the poor bioavailability and improve the anti-inflammatory effect of this polyphenol. Methods: Nanosized lycopenes were encapsulated into biodegradable amphiphillic cyclodextrin and maltodextrin molecules prepared using a high pressure homogenizer at 15,000~25,000 psi. Cell damage was induced by lipopolysaccharides (LPS) in a mouse macrophage cell line, RAW 264.7. The cells were subjected to various doses of free lycopene (FL) and nanoencapsulated lycopene (NEL). RT-PCR was used to quantify the tumor necrosis factor (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$), IL-6, inducible nitric oxide synthase (iNOS), and cyclooxigenase-2 (COX-2) mRNA levels, while ELISA was used to determine the protein levels of TNF-${\alpha}$, IL-$1{\beta}$, and IL-6. Results: NEL significantly reduced the mRNA expression of IL-6 and IL-$1{\beta}$ at the highest dose, while not in cells treated with FL. In addition, NEL treatment caused a significant reduction in IL-6 and TNF-${\alpha}$ protein levels, compared to cells treated with a similar dose of FL. In addition, mRNA expression of iNOS and COX-2 enzyme in the activated macrophages was more efficiently suppressed by NEL than by FL. Conclusion: Overall, our results suggest that lycopene is a potential inflammation reducing agent and nanoencapsulation of lycopene can further improve its anti-inflammatory effect during tissue-damaging inflammatory conditions.

Comparative Analysis of the Community of Culturable Bacteria Associated with Sponges, Spirastrella abata and Spirastrella panis by 16S rDNA-RFLP (16S rDNA-RFLP에 의한 Spirastrella abata와 Spirastrella panis 해면에 서식하는 배양가능한 공생세균 군집의 비교)

  • Cho, Hyun-Hee;Park, Jin-Sook
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.155-162
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    • 2009
  • A cultivation-based approach was employed to compare the culturable bacterial diversity associated with two phylogenetically closely related marine sponges, Spirastrella abata and Spirastrella panis, which have geologically overlapping distribution patterns. The bacteria associated with sponge were cultivated using MA medium supplemented with 3% sponge extracts. Community structures of the culturable bacteria of the two sponge species were analyzed with PCR-RFLP (restriction fragment length polymorphism) based on 16S rDNA sequences. The RFLP fingerprinting of 16S rDNA digested with HaeIII and MspI, revealed 24 independent RFLP types, in which 1-5 representative strains from each type were partially sequenced. The sequence analysis showed >98.4% similarity to known bacterial species in public databases. Overall, the microbial populations of two sponges investigated were found to be the members of the classes; Alphaproteobacteria, Gammaproteobacteria, Firmicutes, and Actinobacteria. The Alphaproteobacteria were predominant in the bacterial communities of the two sponges. Gammaproteobacteria represented 38.5% of bacterial community in S. abata. Whereas only 1.6% of this class was present in S. panis. Bacillus species were dominat in S. panis. Bacillus species were found to be 44.3% of bacterial species in S. panis, while they were only 9.7% in S. abata. It is interesting to note that Planococcus maritimus (8.1%, phylum Firmicutes) and Psychrobacter nivimaris (28.9%, phylum Gammaproteobacteria) were found only in S. abata. This result revealed that profiles of bacterial communities from the sponges with a close phylogenetic relationship were highly species-specific.

Expression and Optimum Production of Cyclodextrin Glucanotransferase Gene of Paenibacillus sp. JB-13 in E. coli (Paenibacillus sp. JB-13 Cyclodextrin Glucanotransferase 유전자의 E. coli 에서의 발현 및 최적 생산)

  • Kim, Hae-Yun;Lee, Sang-Hyeon;Kim, Hae-Nam;Min, Bok-Kee;Baik, Hyung-Suk;Jun, Hong-Ki
    • Korean Journal of Microbiology
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    • v.44 no.1
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    • pp.74-79
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    • 2008
  • The purpose of this study is to clone cgt gene from Paenibacillus sp. JB-13 and to overexpress the protein in E. coli. For this purpose, the cgt gene was amplified from Paenibacillus sp. JB-13 genomic DNA by PCR using degenerate oligonucleotide primers. The sequence analysis results showed that the cgt gene from Paenibacillus sp. JB-13 has 98% homology with the cgt gene of Bacillus sp. To overexpress the protein, the cgt gene was cloned into pEXP7 expression vector and transformed into E. coli. The production of CGTase by recombinant E. coli was optimized under following conditions: 0.5% glucose, 3.0% polypeptone, 0.3% $K_2HPO_4$, 0.5% NaCl, and 7.0 of initial pH, 2.0% of inoculum, $37^{\circ}C$ of culture temperature for 14 hr. And the optimal agitation was found at 0.1 vvm. The synthesis of 2-O-${\alpha}$-D-Glucopyranosyl L-Ascorbic acid (AA-2G) using the CGTase expressed in E. coli was identified as AA-2G by HPLC and HPLC confirmed that treating AA-2G made by cloned CGTase with ${\alpha}$-glucosidase substantially produced AA and glucose.

Cellular Responses to Alcohol in Escherichia coli, Clostridium acetobutylicum, and Saccharomyces cerevisiae (알코올에 대한 Escherichia coli, Clostridium acetobutylicum, Saccharomyces cerevisiae의 반응)

  • Park, Ju-Yong;Hong, Chun-Sang;Han, Ji-Hye;Kang, Hyun-Woo;Chung, Bong-Woo;Choi, Gi-Wook;Min, Ji-Ho
    • Korean Chemical Engineering Research
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    • v.49 no.1
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    • pp.105-108
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    • 2011
  • The increased concern for the security of the oil supply and the negative impact of fossil fuels on the environment, particularly greenhouse gas emissions, has put pressure on society to find renewable fuel alternatives. Compared to the traditional biofuel, ethanol, higher alcohols offer advantage as gasoline substitutes because of their higher energy density and lower hygroscopicity. For this reason, microbial fermentation is known as potential producers for sustainable energy carriers. In this study, bacterial responses including cellular and molecular toxicity were studied in three different microorganisms, such as Escherichia coli, Clostridium acetobutylicum, and Saccharomyces cerevisiae. In this study, it was analyzed specific stress responses caused by ethanol and buthanol using four different stress responsive genes, i.e. fabA, grpE, katG and recA. The expression levels of these genes were quantified by semi-quantitative reverse transcription-PCR. It was found that four genes have shown different responsive patterns when E. coli cultures were under stressful conditions caused by ethanol and buthanol, respectively. Therefore, in this study, the stress responsive effects caused by these alcohols and the extent of each stress response can be analyzed using the expression levels and patterns of different stress responsive genes.

Therapeutic Effects of Yijungtang on Atopic Dermatitis-like Skin Lesions of NC/Nga Mouse Induced by Mite Antigen (이중탕(理中湯)이 Mite Antigen으로 유발된 NC/Nga 생쥐의 아토피 피부염에 미치는 영향)

  • Seo, Hui-Yeon;Han, Jae-Kyung;Kim, Yun-Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.25 no.1
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    • pp.1-27
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    • 2011
  • Objectives: The purpose of this study is to investigate the effects of Yijungtang(YJT) on atopic dermatitis in an in-vitro and in-vivo experiment using a RBL-2H3 mast cells and a NC/Nga atopic dermatitis mouse. Methods: In-vitro experiment, IL-4, IL-13 mRNA expression were evaluated by a real-time PCR, IL-4, IL-13 production by ELISA and transcription factor as GATA-1, GATA-2, NF-AT1, NF-AT2, AP-1 and NF-kB by western blotting. In-vivo experiment, clinical skin score we evaluated by, hematology and Serum total IgE and IgG1 of NC/Nga atopic dermatitis mouse, cytokine level, total number of cell, Immunohistochemical staining and Histological features of auxiliary lymph node(ALN), draining lymph node(DLN), peripheral blood mononuclear cells(PBMCs) and dorsal skin tissue in NC/Nga mouse. Results: YJT decreased IL-4, IL-13 mRNA expression, IL-4, IL-13 production and prominently decreased the expression of mast cell specific transcription factors including GATA-2, NF-AT2, c-Fos and NF-kB. YJT oral administration reduced the levels of skin severity scores. It also decreased the level of inflammatory cytokines such as IL-5, IL-13, histamine and IgE in the serum. It elevated IFN-gamma level in the spleenocyte culture supernatant but decreased. $CD3e^+$, $CD19^+$, $CD4^+$, $CD8^+$, $CD3e^+CD69^+$, $CD11b^+Gr-1^+$, $CCR3^+$ in the PBMCs, $CD4^+$, $CD8^+$, $CD3e^+CD69^+$, $B220^+CD23^+$ in the ALN, $CD4^+$, $CD3e^+CD69^+$ in the ALN and $CD4^+$, $CD11b^+Gr-1^+$ in the dorsal skin. Histological examination showed that infiltration levels of immune cells in the skin of AD-induced NC/Nga mice were much improved by YJT oral administration. Conclusions: The anti-allergic activities of YJT may be mediated by down-regulation of Th2 cytokines, such as IL-4 and IL-13, through the regulation GATA-2, NF-AT2 and NF-kB transcription factors in mast cells. YJT would be regulate molecular mediators and immune cells which are functionally associated with atopic dermatitis induced in NC/Nga mice, and may play an important role in recovering AD symptoms.

Analysis of Monocyte Chemoattractant Protein 1(MCP-1) Polymorphism in Korean Patients with Asthma (한국인 천식환자의 Monocyte chemoattractant protein 1(MCP-1) 유전자 다형성에 대한 분석)

  • Hwang, Woo-Suk;Jeong, Seung-Yeon;Kim, Jin-Ju;Jung, Hee-Jae;Jung, Sung-Ki
    • The Journal of Internal Korean Medicine
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    • v.29 no.1
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    • pp.32-41
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    • 2008
  • Background : Monocyte chemoattractant protein-1(MCP-1), one of the CC chemokines, appears to play a significant role in asthma pathogenesis. It was reported that polymorphism in the MCP-1(-2518 A/G promoter) was associated with asthma in Caucasians, but the association of this polymorphism and asthma patients in the Korean population has not yet been clarified. Objective : We investigated the possible association between 2 polymorphisms (-2518 A/G promoter and Cys35Cys) and asthma patients in a Korean population. Materials and Methods : DNA samples were obtained from 86 Korean asthma patients and 270 healthy controls. MCP-1 genomic variants (-2518 A/G promoter and Cys35Cys polymorphism) were detected by PCR-RFLP. Level of MCP-1 was measured by ELISA for each genotype (n=8) (AA, AG, GG) and allele types of -2518 A/G promoter polymorphism for control subjects. Results : The Cys35Cys polymorphism was associated with asthma patients in Korean population [genotype distribution ($X^{2}=16.011$, P<0.001)]. Comparison of the two groups revealed no detectable differences in genotype and allele frequencies of the -2518 A/G polymorphism. Haplotype frequencies analysis revealed significant difference $(X^{2}=51.70$, P<0.001). MCP-1 serum level of subjects with G genotype of -2518 A/G promoter polymorphism was statistically higher than that with AA genotype (P<0.05). Conclusion : Our data indicate that no association exists between the MCP-1 -2518 A/G polymorphism and asthma susceptibility in the Korean population. However, it is noteworthy that the high prevalence of the -2518 G allele in the Korean population suggests a potentially important ethnic variation in the regulation of MCP-1 production. This variation must be considered in gene-association studies in different ethnic populations.

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