• International Journal of Oral Biology
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• 제46권1호
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• pp.39-44
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• 2021

This study aimed to investigate whether neurotransmitter receptors in the nervous system were also expressed in oral keratinocytes. Expressions of various neurotransmitter receptor genes in immortalized mouse oral keratinocyte (IMOK) cells were examined by reverse transcriptase polymerase chain reaction. IMOK cells expressed calcitonin gene-related peptide (CGRP) receptor subunit genes Ramp1 and Ramp3 and glutamate receptor subunit genes Grina, Gria3, Grin1, Grin2a, and Grin2d. Moreover, IMOK cells expressed Adrb2 and Chrna5 that encode beta 2 adrenergic receptor and cholinergic receptor nicotinic alpha 5 for sympathetic and parasympathetic neurotransmitters, respectively. The expression of Bdkrb1 and Ptger4, which encode receptors for bradykinin and prostaglandin E2 involved in inflammatory responses, was also observed at low levels. Expressions of Ramp1 and Grina in the mouse gingival epithelium were also confirmed by immunohistochemistry. When the function of neurotransmitter receptors expressed on IMOK cells was tested by intracellular calcium response, CGRP, glutamate, and cholinergic receptors did not respond to their agonists, but the bradykinin receptor responded to bradykinin. Collectively, oral keratinocytes express several neurotransmitter receptors, suggesting the potential regulation of oral epithelial homeostasis by the nervous system.

• 대한의생명과학회지
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• 제27권4호
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• pp.283-290
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• 2021

Severe acute respiratory syndrome coronavirus (SARS-CoV2) is a major coronavirus that infects humans with human Coronavirus (HuCoV)-229E, HCoV-OC43, HCoV-HKU1, HCoV-NL63, Severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle east respiratory syndrome coronavirus (MERS-CoV). SARS-CoV2 is currently a global pandemic pathogen. In this study, we developed conventional RT-PCR based diagnostic system for the detection of SARS-CoV2 which is relatively inexpensive but has high stability and a wide range of users. Three conventional RT-PCR primer sets capable of forming specific band sizes by targeting the ORF1ab [232 nucleotide (nt)], E (200 nt) and N (288 nt) genes of SARS-CoV2 were developed, respectively, and it were confirmed to be about 10~100 times higher detection sensitivity than the previously reported methods. In addition, a standard positive control that can generate specific amplicons by reacting with the developed RT-PCR primers and verify the false-positiv from contamination of the laboratory was produced. Therefore, the diagnostic system that uses the RT-PCR method is expected to be used to detect SARS-CoV2.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.53-53
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• 2003

The aim of this study was to determine the interactive effects of BSA and EGF on the viability and development of porcine diploid parthenotes developing in vitro. The addition of 0.1 and 0.4% BSA to the culture medium enhanced the development of 4-cell parthenotes to the blastocyst stage but EGF had no effect. However, while BSA also increased cell numbers, it did so only when EGF was also present. Either agent on its own had no effect. Similarly, apoptosis in the blastocysts was not influenced by either agent on its own but was reduced when both BSA and EGF were present. Furthermore, semi-quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) revealed that EGF enhanced the mRNA expression of BclxL in the presence of 0.4% BSA but BSA and EGF alone had no effect. EGF and/or BSA did not influence Bak gene expression in the blastocyst stage parthenotes. These results suggest that BSA has both beneficial and detrimental effects on the viability of porcine diploid parthenotes developing in vitro and that exogenous EGF may block some of the detrimental effects of BSA, possibly by inhibiting the BSA-induced apoptosis by increasing Bcl-xL expression. This results in a net increase in cell numbers in porcine diploid parthenotes developing in vitro.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.47-47
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• 2003

The present study was conducted to determine the expression of the antioxidant enzyme(CuZn-SOD, Mn-SOD and GPX and apoptosis gene(caspase-3) for in vitro culture in in vitro maturation and in vitro fertilization(IVM/IVF) embryos in porcine. Porcine embryos derived from IVM/IVF were cultured in NCSU23 medium under 5% $CO_2$ in air at 38.5$^{\circ}C$. The patterns of gene expression for several antioxidant enzyme and apoptosis genes during preimplantion porcine embryo development were examined by the modified semi-quantitative single cell reverse transcriptase- polymerase chain reaction (RT-PCR). Preimplantation porcine embryos produced by IVM/IVF have expressed mRNAs for CuZn-SOD and GPX, whereas transcripts for Mn-SOD have not detected at any developmental stages. Expression of caspase-3 mRNA was detected at 2 cell, 8 cell, 16 cell and morula stages. The fas ligand transcripts were detected in porcine blastocyst. These results suggest that various antioxidant enzymes and apoptosis genes play crucial roles in in vitro culture of porcine IVM/IVF embryos.

• Journal of Veterinary Science
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• 제21권5호
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• pp.71.1-71.10
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• 2020

Background: Feline calicivirus (FCV) is a major and highly infectious pathogen in cats worldwide. However, there have been limited studies about the status of FCV infections in Korea. Objectives: To investigate the current status of FCV infections in stray cats in Korea. Methods: A novel reverse transcription polymerase chain reaction (RT-PCR) assay was developed based on the conserved nucleotide sequences of reported FCV strains. Field swab samples were collected from 122 cats (2 hospital admitted cats and 120 stray cats) in 2016 and 2017. All the samples were tested by virus isolation and 2 different RT-PCRs, including the novel RT-PCR, for the detection of FCV. Results: The novel RT-PCR assay showed no cross-reactivity to the nucleic acids of the other feline pathogens tested, and the limit of detection was calculated as 10⁰ TCID₅₀/mL based on an in vitro assessment. The novel RT-PCR assay detected 5 positive samples from the 122 field samples, which showed perfect agreement with the results of the virus isolation method. In contrast, another RT-PCR assay used in a previous study in Korea detected no positive samples. The prevalence of FCV infection in stray cats was 2.5% (3/120) based on the results of virus isolation and the novel RT-PCR assays. Conclusions: The current study is the first report of the detection and prevalence of FCV in stray cats in Korea. The novel RT-PCR assay developed in this study showed high sensitivity and specificity, which indicates a useful diagnostic assay to identify FCV infection in cats.

• Korean Journal of Acupuncture
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• 제31권2호
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• pp.56-65
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• 2014

Objectives : The purpose of this study is to compare the effects of laser acupuncture to the 830 nm on the five transport points with the spleen meridian for treatment to intestinal disease in rat with dextran sulfate sodium(DSS)-induced colitis. Methods : Colitis was induced by DSS for 20 days. The laser therapy on the five transport points of spleen meridian (Laser Well Point-SP1(L-WE), Laser Brook Point-SP2(L-BR), Laser Stream Point-SP3(L-ST), Laser River Point-SP5(L-RI) and Laser Sea Point-SP9(L-SE) was practiced twice a week for 5 times. Colon length was measured using a measuring point. Histological evaluation of colitis was conducted by hematoxylin and eosin(H&E) staining. Reverse transcription polymerase chain reaction(RT-PCR) was determined using western blotting and quantitative reverse-transcriptase polymerase chain reaction, respectively. Results: Colon length increased significantly L-BR and L-ST points after 5 times of therapy. Damage to the colonic mucosa is an integral feature of the DSS model, so control colonic mucosa tissue was damaged in the areas of ulceration resulting in complete epithelial loss. However histological damage decreased on the epithelial lining at all points. Cyclooxygenase(COX)-2 concentrations decreased in all points groups and Interferon(IFN)-${\gamma}$ increased in L-WE, L-BR, L-RI and L-SE points but L-ST was decreased when compared with control. White blood cell(WBC) and neutrophils(NE) decreased after the fifth acupuncture on the all points. But hemoglobin(HGB) increased after the fifth acupuncture on the L-WE, L-BR, L-ST and L-RI points. Also Mean corpuscular hemoglobin(MCH) and Mean corpuscular hemoglobin concentration(MCHC) decreased after the fifth acupuncture on the all points. Conclusions: The present study indicated that five transport points of the spleen meridian can prevent the development of DSS-induced colitis in rat. Thereby suggesting that should be available for decreasing DSS-induced inflammation in a colonic mucosa of tissue.

• 동의생리병리학회지
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• 제23권6호
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• pp.1305-1313
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• 2009

Saengjihwangeum-ja (SJHEJ) was recorded in DongEuiBoGam as being able to be used for treatment of Sogal whose concept had been applied to Diabetes Mellitus (DM). Modification of proteins by long term circulation of glucose leads to the formation of advanced glycation end product(AGE). Recent immunological studies demonstrated that ligation of AGE play an important role in the development of diabetic complications including atherosclerosis, which includes activation, adhesion, and migration of monocytes. Also, AGE and Maillard reaction product(MRP) could augment monocyte inflammatory responses via ligation of AGE receptor. In this study, the effects of SJHEJ extracts on the expression of inflammatory response-related genes such as tumor necrosis factor-$\alpha$, monocyte chemoattractant protein-1, interferon-g-inducible protein-10, and cyclooxygenase-2 in the human monocyte cell line, THP-1 cells. Reverse transcriptase-polymerase chain reaction revealed that SJHEJ had inhibitory effects on the expression of the TNF-a, MCP-1, IP-10, COX2, IL-1b genes in MRP-induced THP-1 cells. Treatment with SJHEJ had reduced reactive oxygen production in THP-1 cells stimulated by MRP. These inhibitory effects might be exerted via prevention of oxidative stress in activated monocytes. In addition, radical scavenging activity of SJHEJ was increased. These results suggest that SJHEJ has a beneficial effects for improve diabetic vascular complication.

• Journal of Microbiology and Biotechnology
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• 제25권3호
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• pp.343-352
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• 2015

H9 is an ethanol extract prepared from nine traditional/medicinal herbs. This study was focused on the anticancer effect of H9 in non-small-cell lung cancer cells. The effects of H9 on cell viability, apoptosis, mitochondrial membrane potential (MMP; ${\Delta}\psi_{m}$), and apoptosisrelated protein expression were investigated in A549 human lung cancer cells. In this study, H9-induced apoptosis was confirmed by propidium iodide staining, expression levels of mRNA were determined by reverse transcriptase polymerase chain reaction, protein expression levels were checked by western blot analysis, and MMP (${\Delta}\psi_{m}$) was measured by JC-1 staining. Our results indicated that H9 decreased the viability of A549 cells and induced cell morphological changes in a dose-dependent manner. H9 also altered expression levels of molecules involved in the intrinsic signaling pathway. H9 inhibited Bcl-xL expression, whereas Bax expression was enhanced and cytochrome C was released. Furthermore, H9 treatment led to the activation of caspase-3/caspase-9 and proteolytic cleavage of poly(ADP-ribose) polymerase; the MMP was collapsed by H9. However, the expression levels of extrinsic pathway molecules such as Fas/FasL, TRAIL/TRAIL-R, DR5, and Fas-associated death receptor were downregulated by H9. These results indicated that H9 inhibited proliferation and induced apoptosis by activating intrinsic pathways but not extrinsic pathways in human lung cancer cells. Our results suggest that H9 can be used as an alternative remedy for human non-small-cell lung cancer.

• 한국동물학회지
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• 제33권1호
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• pp.35-44
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• 1990

생쥐 악하선으로부터 분리한 전 RNA를 poly(U)-sepharose chromatography와 sucrose linear density gradient centrifugation 방법으로 레닌 mRNA를 분리하여 in vitro translation과 immunoprecipitation에 의하여 레닌 mRNA를 확인하였다. 레닌 mRNA로부터 레닌 cDNA를 합성하여 EcoRI inker를 이용하여 pUC19에 삽입시켰고, Taq, polymerase를 이용한 PCR방법으로 합성한 레닌 cDNA는 pUC19의 Hinalll 부의에 삽입하여 재조합 plamid를 각각 작성하였다. 이것을 JM103에 형질전환시켜 레닌 유전자 발현을 유도하여 45,000의 분자량을 갖는 레닌을 얻었으며 이 레닌 단백은 토끼으 혈압을 850115mmHg에서 115-140mmHg로 증가시키는 생리 활성을 나타냈다. 토끼의 신장 DNA를 EMBL3 phage에 삽입시켜 genomic library를 작성한 후, 레닌 cDNA로부터 합성한 probe로 plaque hybridization시켜 genomic 유전자를 갖는 재조합 phage를 분리하였다.

• KSBB Journal
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• 제16권6호
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• pp.562-567
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• 2001

Styrene oxide 계열의 라세믹 에폭사이드 기질에 대한 입체선택적 가수분해능이 우수한 Aspergillus nigerr계열의 생촉매를 선발하였고, A.niger LK 유래의 EHase의 기질 특이성을 분석하였다. A. niger LK의 EHase는 benzene ring에 oxirane ring이 직접 연결되어 있는 styrene oxide, p-nitrostyrene oxide 기질에 대해서는 (R)-이성질체, benzene ring과 oxirane ring사이에 ether 등의 연결 chain이 있는 기질에 대해서는 (S)-이 성질체에 대한 입체선택적 가수분해능이 우수하였다. A niger LK의 EHase 유전자를 RT-PCR 방법으로 클로닝하였고, sequencing을 통해 다른 미생물 유래의 EHase와의 sequence identity 분석 등을 통해 특성을 분석하였다. Yeast 유래의 EHase와는 32% 수준의 sequence identity를 보였으며, Agrobacterisum, Corynebacterium 등의 박테리아 유래 EHase와는 identity가 매우 낮은 특성을 보였다. E. coli 숙주에서 발현된 재조합 EHase의 활성은 라세믹 에폭사이드 기질에 대한 입체선택적 가수분해 반응을 통해 확인할 수 있었다. 클러닝된 EHase의 보다 효율적인 발현 연구가 필요하며, 이러한 재조합 EHase는 고부가가치 광학활성 에폭사이드 제조를 위한 생물전환공정 시스템의 생촉매로 응용될 수 있을 것으로 기대된다.