• Korean Journal of Animal Reproduction
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• v.18 no.3
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• pp.199-206
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• 1994

We present here a new PCR-based cloning technique that allows the different PCR products during mouse embryogenesis. Recently, mRNA differential display described by Liang & Pardee (Science 257, 1992) and re-confirmed by Zimermann & Schultz (PNAS 91,1994). This method will detect the appropriate changes in the temporal patterns of expression or in the transition from maternal control to zygotic control as well as the functional difference of embryo with polyspermy or monospermy, the difference of expression between successfully hatched blastocyst and blastocyst failed to hatching, response to agents, and cell cycle regulation. By this methods, we have cloned an eDNA, which showed mouse 2 cell specific expression. Genomic DNA digested with EcoRI showed approximately 15 kb and then showed higher expression in fetal liver rather than adult liver. Furthermore, this gene is likely to have 2 mRNA by alternative splicing.

• Journal of Pharmacopuncture
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• v.4 no.1
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• pp.5-21
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• 2001

The influence of electroacupuncture (EA), a traditional Chinese medical treatment, on type Ⅱ collagen-induced arthritis (CIA) was examined in DBA/1J mice in vivo. Mice were immunized intradermally twice at the 3-week interval with bovine type Ⅱ collagen(C Ⅱ). EA stimulation, begun on the 21 simultaneously with the second immunization, was applied at the acupoint equivalent to GV4 three times a week for 3 weeks. The results showed that EA delayed the onset, attenuated the severity of arthritis, and reduced the anti-collagen antibody level. Furthermore, we investigated the impact of EA on the productions of endogenous $interleukin-1{\Beta}$ (IL-1 beta) and prostaglandin E2 (PGE2), and the levels of IL-1 beta mRNA in splenocytes and synovial tissues from C Ⅱ immunized mice on the 45 and cyclooxygenase-2 (COX-2) mRNA in lipopolysaccharide (LPS)-stimulated macrophages of normal mice by using reverse transcriptase-polymerase chain reaction (RT-PCR). EA stimulation significant inhibited the concentrations of splenic endogenous IL-1 beta and serum PGE2. The expression of IL-1 beta mRNA in spleen cells was obviously down-regulated and that in synovial tissues was modestly affected by EA. COX-2 mRNA was highly expressed in cultured peritoneal macrophages when stimulated with LPS. Previous treatment with EA also reduced LPS-stimulated induction of COX-2 mRNA. These data suggest that EA has an inhibitory effect on murine CIA, and the partial mechanism of its therapeutic result may be attributed to inhibiting the productions of IL-1 beta and PGE2 by suppression the IL-1 beta and COX-2 gene activations.

• Korean Journal of Animal Reproduction
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• v.25 no.4
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• pp.381-388
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• 2001

The present study was designed to obtain porcine growth hormone binding protein (pGHBP) improved biological activation as derived mutation in binding site with growth horlnone (GH). A 756 bp of fragment encoding the extracellular domain of pGHBP gene was cloned from the total RNA of porcine fat tissue by reverse transcriptase polymerase chain reaction (RT-PCR) and created mutation in positions 26 and 122 using site-directed mutagenesis method. Position 26 is one and it is near to get on five potential N-linked glycosylation sites located in the extracellular domain of porcine growth hormone receptor known to have a direct influence on combination with GH. Position 122 is known as one of conformational epitope in bovine. It was over-expressed in E. coli using pET-32(c) expression vector and precisely purified by S-protein agarose and enterokinase. In our results, we was obtained pmGHBP of 30 kDa. It suggests to study the effects of the pmGHBP on cell proliferation in vitro and growth rate in vivo after administration.

• Tuberculosis and Respiratory Diseases
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• v.68 no.6
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• pp.350-353
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• 2010

Here we report the first fatality caused by H1N1 influenza virus infection with acute respiratory distress syndrome in Korea. A 55-year-old man presented at our emergency department with dyspnea, fever, diffuse myalgia and malaise. Bilateral lung air-space consolidation was detected on his initial chest radiograph combined with severe hypoxemia. He was supported by mechanical ventilation and treated with antibiotics. A nasopharyngeal aspirate was positive for influenza A rapid antigen and oseltamivir was started on day 3 of admission. The nasal swab sample was positive for influenza H1N1 virus by real-time reverse-transcriptase polymerase chain reaction. Despite aggressive treatment, he had refractory hypoxemia and uncontrolled septic shock. On day 5 of admission he went into cardiac arrest and expired.

• Journal of Aquaculture
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• v.13 no.1
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• pp.79-85
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• 2000

The effects of estrogen and androgens on Vg gene expression were examined in primary hepatocyte culture and livers of the immature male trout. Specific primers of Vg cDNA were designed with already reported Vg gene nucleotide sequences. PCR product was sequenced and verified with Vg cDNA of rainbow trout. Total RNA was extracted from the cultured hepatocytes and livers of steroid-treated rainbow trout and then it was analyzed by reverse transcriptase- polymerase chain reaction (RT-PCR) analysis. The Vg mRNA and Vg protein synthesis were increased in rainbow trout in vivo and in vitro with E$_2$ and methyltestosterone (MT) There were dose and time-related effects of E$_2$ and MT on vitellogesis. Androgens such as progesterone androsterone and testosterone also stimulated Vg mRNA expression in vitro. The results show that androgens as well as E$_2$ can induce expression of Vg mRNA in trout in vivo and in vitro.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.28 no.4
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• pp.51-61
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• 2015

Objective : This research has been conducted to comparative analysis of anti-inflammatory effects to suggest a usefulness of ethanol extracts fromGentianae sino-ornata(GSO) as a functional material for inflammatory activities.Methods : Cell viability was measured by neutral red (NR) assay, and nitric oxide (NO) production in RAW264.7 cells was monitored by measuring the nitrite content in culture medium. The expressions of cyclooxigenase-2 (COX-2) was determined by western blot analysis, and Inducible nitric oxide syntase (iNOS) and cytokine were determined by reverse transcriptase-polymerase chain reaction (RT-PCR).Results : When the GSO extract was added the concentration of 5-20 ㎍/㎕, the viability of cells was maintained 90% or more at all levels. NO production was suppressed by the treatment of GSO in LPS-stimulated RAW264.7 cells. GSO inhibited the expression of iNOS, COX-2, IL-1βand IL-6 in LPS-stimulated RAW264.7 cells.Conclusions: From this results, we consider that GSO can be a useful therapeutic and preventive approach to various inflammatory diseases as a functional material for inflammatory activities.

• Journal of Sasang Constitutional Medicine
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• v.29 no.2
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• pp.136-153
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• 2017

Objectives This experimental study was designed to investigate the effect of Phaseolus angularis shell on metabolic syndrome. Methods Each 5 C57BL/6J mice were randomly assigned to normal diet group, high-fat diet(HFD) control group, high-fat diet plus 15.6 mg/kg/day of Orlistat(HFD-Orlistat) group, high-fat diet plus 100mg/kg/day of Phaseolus angularis shell extract(HFD-PAS_E) group. Weight, the blood chemical and hematologic parameter was med. The mRNA expression was assayed through Reverse transcriptase polymerase chain reaction(RT-PCR). Results In HFD-PAS_E group, the body weight gain, weight of liver, and the level of LDL-Cholesterol were significantly decreased and the level of HDL-Cholesterol were significantly increased. The size of adipocyte in HFD-PAS_E group was smaller than HFD group's. In HFD-PAS_E group, the expression of leptin, PPAR-${\gamma}$, AP2/FABP4 mRNA in liver adipocyte tissue was decreased, the expression of Adiponectin, UCP-2 mRNA in liver adipocyte tissue was increased and the expression of Leptin, C/EBP-a, AP2/FABP4 mRNA in epididymal adipocyte tissue was decreased. Conclusion These results suggest that Phaseolus angularis shell has inhibitory effects on metabolic syndrome by reducing the body weight and the levels of lipid contents in high-fat-diet induced obese mice.

• Animal cells and systems
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• v.1 no.4
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• pp.589-594
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• 1997

The neuronal voltage-sensitive calcium channels (VSCCs) are multisubunit complexes consisting of $\alpha_1,\;\alpha_2-\delta$ and $\beta$ subunits. Heterologous expression and biochemical studies have shown that the activity of VSCCs is regulated by their $\beta$ subunits in a $\beta$ subunit isoform-specific manner. To elucidate the $\beta$ subunit identity of the P/Q-type calcium channel encoded by an $\alpha_{1A}$ subunit, which is exclusively expressed in the Purkinje and granule cell of the cerebellum, we have examined the spatial and temporal expression patterns of $\beta$ subunits and compared them with those of $\alpha_{1A}$ subunit in the developing rat cerebellum. Reverse transcriptase- polymerase chain reaction (RT-PCR) and Northern blot analysis have shown that $\beta_4$ subunit mRNA was prominently expressed in the cerebellum and much more abundant than any other distinct $\beta$ subunits. RNase protection assay has further demonstrated that the expression of $\alpha_{1A}$ and $\beta_4$ subunits increased during cerebellar development, while the amount of $\beta_2$ and $\beta_3$ mRNAs did not significantly change. In addition, a $\beta_4$ transcript was present in cultured cerebellar granule cells, but not in astrocyte cells, and the level of $\beta_4$ mRNA expression increased gradually in vitro seen as in vivo. Based on the spatial and temporal expression patterns of $\beta_4$ subunit, we conclude that $\beta_4$ may predominantly associate, but probably not exclusively, with the $\alpha_{1A}$ subunit in rat cerebellar granule cells.

• Korean Journal of Veterinary Service
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• v.40 no.2
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• pp.101-105
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• 2017

The purpose of this study was to investigate prevalence and antimicrobial resistance patterns of Campylobacter spp. from duck feces in northern area of the Gyeongnam province, Korea. Samples of 121 duck feces were taken from April to December 2014 for this survey. Samples were examined by bacteria isolation and reverse transcriptase-polymerase chain reaction assay for detection of Campylobacter spp. Campylobacter were isolated in 37 samples (30.6%). Among these samples, C. jejuni and C. coli were isolated in 35 samples and 2 samples, respectively. Minimum inhibitory concentration (MIC) test is performed to investigate antimicrobial resistance patterns of Campylobacter spp. C. jejuni were resistant to ciprofloxacin (85.7%), nalidixic acid(82.9%), tetracycline (77.1%), gentamicin (57.1%), azithromycin (40.0%), clindamycin (34.3%), erythromycin (22.9%), and florfenicol (8.6%). These data support a database of pollution and antimicrobial resistance of Campylobacter spp. from duck feces and provide a basic information of reducing the secondary damage of antibiotic misuse.

• Korean Journal of Veterinary Research
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• v.46 no.3
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• pp.235-248
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• 2006

The VP2 full gene of Korean infectious bursal disease virus(IBDV) strain, SH/92, three attenuated vaccine strains, Bur706, Bursine-2 and CEV/AC strains, were amplified by reverse transcriptase-polymerase chain reaction and sequenced and compared with published VP2 gene sequences of IBDVs. The VP2 nucleotide sequence similarity between SH/92 and three vaccine stains was 95.6~96.5% whereas the nucleic acid similarity among three vaccine strains was 97.5~98.5%. The amino acid sequence similarity of VP2 of SH/92 compared with three vaccine strains was between 94.4 and 97.6% while the amino acid similarity among three vaccine strains was between 97.4 and 98.4%. The amino acid similarity between SH/92 and classical virulent strain, 52/70 and STC strain was 96.4 and 96.5%, respectively. The serine-rich heptapeptide was conserved in CEVAC and Bursine-2 as well as SH/92 but not in Bur706. The phylogenetic tree developed from amino acid sequences showed that SH/92 was categorized with vv IBDVs(HK46, OKYM, KKI, UPM94/273, SH95) in one branch while three vaccine strains were catagorized with STC strain in the other branch.