• The Journal of Korean Medicine
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• v.22 no.2
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• pp.109-119
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• 2001

Objective : There are many reports that acupuncture has thermoregulatory effects on human and animals. To investigate the effect and mechanism of antipyretic action of acupuncture, we observed the body temperature and cytokine expressions in the hypothalamus of rats. Methods : Lipopolysaccharide (LPS, i.p., 2.5mg/kg) was injected to conscious rats (Sprague-Dawley, male, n=4l) to cause hyperthermia and simple needling (stainless steel, 0.25 mm o.d., 5 mm insertion for 10 sec with no manipulation) was performed bilaterally with the measurement of rectal temperature. Next, we sacrificed rats to remove brain and determined the level of mRNA for interleukin-6 (IL-6), $interleukin-1{\beta}{\;}(IL-1{\beta})$, interleukin-2 (IL-2) and $interferon-{\gamma}{\;}(IFN-{\gamma})$ in the hypothalamus by using reverse transcriptase-polymerase chain reaction (RT-PCR). Resul1s : Needling on forepaw (acupoint HT8) and needling on hindpaw (acupoint BL66 and acupoint LR2) significantly inhibited LPS-induced fever of rats (P<0.01, 10 min after treatment respectively), but same treatment on proximal tail (non-acupoint) did not cause any change on fever. The levels of IL-6 and $IL-1{\beta}$ mRNA in the hypothalamus was significantly enhanced by LPS-injection, while the level of IL-6 and $IL-1{\beta}$ mRNA was markedly reduced after treatment on BL66 (P<0.01). Treatment on forepaw reduced it slightly, but not significantly. Equivalent stimulation on proximal tail did not cause any changes. Conclusions : Our results indicate that acupuncture stimulation on various body parts has differential thermoregulatory effects on LPS-induced fever of rats with site-specificity. And, we suggest that its antipyretic action might be accompanied with the suppression of hypothalamic production of pro-inflammatory cytokine of immune system, IL-6 and $IL-1{\beta}$.

• Journal of Physiology & Pathology in Korean Medicine
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• v.26 no.3
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• pp.273-280
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• 2012

Scutellaria baicalensis Georgi, which has been known to be able to clear away heat and remove dampness, was used for febrile disease. It is now clear that Advanced glycation end products (AGEs) play major roles in the pathogenesis of diabetic complications such as atherosclerosis. In this study, we examined whether Scutellaria baicalensis Georgi suppress the AGE mediated inflammatory responses in the THP-1 cells. AGE treatment increased the gene expression of pro-inflammatory cytokines such as tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$), monocyte chemotactic protein-1 (MCP-1) and cyclooxygenase-2 (COX-2). Reverse transcriptase-polymerase chain reaction and Western blot analysis revealed that S. baicalensis had inhibitory effects on the expression of pro-inflammatory genes and protein levels in AGE-treated THP-1 cells. S. baicalensis had also reduced the production of ROS in the AGE-treated THP-1 cells. These results suggest that S. baicalensis has inhibitory effects for the development of diabetic vascular complication.

• Journal of Genetic Medicine
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• v.1 no.1
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• pp.51-56
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• 1997

Spermatogenesis is known to be regulated by a number of genes and several factors such as hormones, growth factors, cytokines and others. This study was done to evaluate the relationship between HSPs and DAZ genes in human spermatogenesis; we observed the expression pattern of HSP gene in azoospermia men with DAZ gene that regulated the gene expression related with human spermatogenesis. RT-PCR method was used to detect DAZ, HSP70A, and HSP70B transcripts in all RNA samples. Total RNA was extracted from 21 testis tissues using TRIZOL reagent. cDNAs were synthesized with reverse transcriptase, AMV. All PCR reaction were performed on a PCR themocycler with DAZ, HSP70A, and HSP70B-specific primers. Semen analysis, karyotyping and testis histology were performed. DAZ gene, known as a candidate gene of azoospermia factor(AZF), was deleted in 2 of 21 patients. To evaluate the only effects of HSPs in this patients, 2 DAZ deleted cases were removed. We observed the mRNA of HSP70B in 5 whereas none could be seen with regard to HSP70A. Furthermore, the sperm of these 5 men were discovered to be immature. In conclusion, HSP70B as well ad DAZ gene seem to be involved causing spermatogenic failure. We suggest that HSP70B plays an important role in spermatogenesis and it is one of factors induced sperm maturation in human.

• The Plant Pathology Journal
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• v.15 no.6
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• pp.335-339
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• 1999

Citrus tristeza virus(CTV), an aphid-borne closterovirus, is one of the most destructive pathogens of citrus. It has caused rapid decline in growth, stem pitting and death in citrus trees. A reverse transcriptase polymerase chain reaction (RT-PCR) assay was developed for detection of CTV and investigation of the CTV infection status of citrus and its related cultivars in Cheju island. For RT-PCR based CTV detection, primers were designed to amplify 670bp of coat protein gene. A screening test for CTV in citrus cultivars was conducted from March to July in 1999. Seventy individual citrus trees representing 9 species of 3 genera were tested. The infection rates of CTV for leaves from the years or older trees of late maturing citrus varieties such as Yuzu (C. junos Sieb. ex Tanaka), Navel orange (C.sinensis Osbeck), Kiyomitanger (C. unshiu x C. sinensis), and Shiranuhi ((C. unshiu x C. sinensis) x C. reticulata) were 100%, 80%, 60%, and 60% respectively. The CTV infection rates in Early satsuma mandarins such as 'Miyagawa Early' Satsuma mandarins (C. unshiu Marc. var. Miyagawa) and 'Okitsu Early' Satsuma mandarins (C. unshiu Marc. var. Okitsu) were 100%, and 60%, respectively. CTV was not detected in Cheju native Dangyooja (C. unshiu Marc. var. Osbeck), Trifoliate orange (Poncirus trifoliata) and Kumquat (Fortunella margarita Swingle). In conclusion, RT-PCR assay can be successfully applied to the detection of CTV in citrus trees.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.4
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• pp.576-584
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• 2002

Bovine PAS-4 is an integral membrane glycoprotein expressed in mammary epithelial cells. Complementary DNA (cDNA) cloning of PAS-4 was performed by reverse-transcriptase polymerase chain reaction (RT-PCR) with oligonucleotide probes based on it's amino terminal and internal tryptic-peptides. The cloned PAS-4 cDNA was 1,852 nucleotides (nt) long and its open reading frame (ORF) was encoded 1,413 base long. The deduced amino acid sequence indicated that PAS-4 consisted of 471 amino acid residues with molecular weight of 52,796, bearing 8 potential N-glycosylation sites and 9 cysteine residues. Partial bovine CD36 cDNA from liver also was sequenced and the homology of both nucleotide sequence was 94%. Most of the identical amino acid residues were in the luminal/extracellular domains. Contrary to PAS-4, bovine liver CD36 displays 6 potential N-glycosylation sites, which were located, except for those at positions 101 and 171, at same positions as PAS-4 cDNA. Cysteine residues of PAS-4 and CD36 were same at position and in numbers. Northern blot analysis showed that PAS-4 was widely expressed, although its mRNA steady-state levels vary considerably among the analyzed cell types. PAS-4 possessed hydrophobic amino acid segments near the amino- and carboxyl-termini. Two short cytoplasmic tails of the amino- and carboxyl-terminal ends constituted of a 5-7 and 8-11 amino acid residues, respectively.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.12
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• pp.1830-1836
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• 2003

Studies using natural resistance-associated macrophage protein (NRAMP) identification indicated that cattle could be selected for immunity. Several studies performed on intracellular organisms such as Mycobacterium, Salmonella, Brucella and Leishmania in human and mouse revealed that resistance against these bacteria was dependent on high activity of NRAMP1 in macrophages. However, hardly any researches have been done on Staphylococcus aureus in bovine mastitis, which is an intracellular organism and the main cause of bovine mastitis. The objectives of this study were to establish reverse transcriptase polymerase chain reaction (RT-PCR) methods, through which NRAMP1 mRNA expression could be compared and analyzed between mastitis-resistant and -susceptible cows. NRAMP1 gene and its expression were investigated using 20 cows (Holstein Friesian) in Korea. Cows were evenly split into two groups, with and without histories of clinical mastitis. Equivalent numbers of cows were randomly selected from each group. Monocytes were isolated from the bovine peripheral blood of each selected cows and activated with lipopolysaccharide (LPS). mRNA was separated from the monocytes and cDNA of NRAMP1 was synthesized and amplified using RT-PCR with amplification of $\beta$-actin as a control. The difference in NRAMP1 expressions of mastitis-resistant (n=10) and -susceptible (n=10) Holstein cows was analyzed. Results demonstrate that resistant cows produced more NRAMP1 mRNA than the susceptible ones, and ratios of NRAMP1:$\beta$-actin expression were higher in resistant cows with or without LPS activation. Therefore, this study could be applied to select bovine mastitis resistant cows before infection based on the expression of NRAMP1.

• Journal of Ginseng Research
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• v.35 no.4
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• pp.479-486
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• 2011

Atopic dermatitis (AD) is an allergic, inflammatory skin disease characterized by chronic eczema and mechanical injury to the skin, caused by scratching. Korean red ginseng (RG) has diverse biological activities, but the molecular effects of RG on allergic diseases, like AD, are unclear. The present study was designed to investigate whether RG inhibits 1-chloro-2,4-dinitrobenzene (DNCB)-induced AD in a mouse model. DNCB was applied topically on the dorsal surface of Balb/c mice to induce AD-like skin lesions. We observed the scratching behavior and examined the serum IgE level and interleukin (IL)-4 and IL-10 in splenocytes compared with dexamethasone. We also evaluated the DNCB-induced mitogen-activated protein kinases (MAPKs), NF-${\kappa}B$, and Ikaros activities after RG treatment using reverse transcriptase-polymerase chain reaction, Western blotting, and ELISA. Our data showed that the topical application of RG significantly improved the AD-like skin lesions and scratching behavior. RG decreased not only the mRNA expression of IL-4 and IL-10, but also the secretion of IL-4 protein and serum IgE in mice. Additionally, RG treatment decreased the DNCB-induced MAPKs activity and subsequent Ikaros translocation irrespective of NF-${\kappa}B$. We suggest that RG may be useful as a therapeutic nutrition for the treatment of AD.

• Journal of Ginseng Research
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• v.33 no.3
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• pp.212-218
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• 2009

Triterpenoid saponins were synthesized in Panax ginseng C.A. Meyer via the isoprenoid pathway by cyclization of 2,3-oxidosqualene to give primarily oleanane (beta-amyrin) or dammarane triterpenoid skeletons. The triterpenoids are backbone and undergoes various modifications (oxidation, substitution and glycosylation), mediated by cytochrome P450 (CYP)-dependent monooxygenases, glycosyltransferase and other enzymes. This is likely to be due in part to the complexity of the molecules and the lack of pathway intermediates for biochemical studies. A cDNA clone encoding a putative CYP gene was isolated from flower bud of ginseng and transformed into the plant(Nicotiana tabacum cv. Xanthi) and confirmed by PCR analysis. The CYP gene (PgCYP) contained an open reading frame(ORF) encoding mature protein of 500 amino acids. The expression of PgCYP were investigated in transgenic tobacco by reverse transcriptase-polymerase chain reaction (RT-PCR).

• Plant Biotechnology Reports
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• v.5 no.1
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• pp.53-60
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• 2011

Artemisinin is effective against both chloroquine-resistant and -sensitive strains of Plasmodium species. However, the low yield of artemisinin from cultivated and wild plants is a serious limitation to the commercialization of this drug. Optimization of artemisinin yield either in vivo or in vitro is therefore highly desirable. To this end, we have overexpressed the 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (HMGR) gene (hmgr) from Catharanthus roseus L. in Artemisia annua L. and analyzed its influence on artemisinin content. PCR and Southern blot analyses revealed that the transgenic plants showed stable integration of the foreign hmgr gene. The reverse transcriptase-PCR results suggested that the hmgr was expressed at the transcriptional level in transgenic lines of Artemisia annua L., while the high-performance liquid chromatography analysis showed that artemisinin content was significantly increased in a number of the transgenic lines. Artemisinin content in one of the A. annua transgenic lines was 38.9% higher than that in non-transgenic plants, and HMGR enzyme activity in transgenic A. annua L. was also higher than that in the non-transgenic lines.

• Korean Journal of Veterinary Research
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• v.50 no.2
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• pp.139-143
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• 2010

A 2-month-old male Jeju native black calf with respiratory distress was died and requested to the Veterinary Pathology Laboratory of Jeju National University for diagnosis. Grossly, lungs were focally attached to the pleura and heart with fibrin. Purple red sublobar consolidations were distributed in both apical and cardiac lobes of lungs. Histopathologically, diffuse severe bronchointerstitial pneumonia characterized by multifocal necrotizing bronchiolitis, formation of numerous multinucleated syncytial cells in bronchiolar and alveolar lumens, and diffuse alveolar wall thickening were observed in lungs. Eosinophilic intracytoplasmic inclusions were observed in bronchiolar epithelial cells and syncytial cells. According to reverse transcriptase polymerase chain reaction (RT-PCR), bovine respiratory syncytial virus (BRSV) was detected in the lung of calf. Based on the histopathologic findings and RT-PCR, this calf was diagnosed as BRSV infection. In our best knowledge, this is the first case of BRSV infection in Jeju native black calf.