• Journal of Pharmacopuncture
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• v.7 no.3
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• pp.5-25
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• 2004

Objectives : To observe changes in the serum proteins before and after intravenous injection of wild ginseng herbal acupuncture. Methods : Blood was collected before and after the administration of wild ginseng herbal acupuncture and only the serum was centrifuged. Then differences in the spots on the scanned image after running 2-Dimensionl electrophoresis were located and conducted mass analysis and protein identification. Results : Following results were obtained from the comparative analysis of serum proteins before and after the administration of wild ginseng herbal acupuncture. 1. 28 spots were identified before and after the administration. 2. In confirming manifestation degree, spots with more than two-times increase were 204, 803, 1505, 2205, 3105, 7104, 9001 spots, with more than one-time increase were 1101, 1302, 2013, 3009, 3010, 4002, 4009, 6706, 7103, 8006, 8101, and spots with decrease were 205, 801, 3205, 5202, 6105. 3. After conducting protein identification, proteins 205, 804, 1302, 4009, 6105, 6106 are unidentified yet, and 1101 is unnamed protein. Protein 204 is identified as complement receptor CR2-C3d, 801 as YAP1 protein, 803 as antitrypsin polymer, 1505 as PRO0684, 2013 and 3010 as proapolipoprotein, 2205 as USP48, 2403 as vitamin D binding protein, 3009 as complement component 4A preprotein, 3105 as immunoglobulin lambda chain, 3205 as transthyretin, 4002 as Ras-related protein Ral-A, 4204 as beta actin, 5202 and 7104 as apolipoprotein L1, 6704 as alpha 2 macroglobulin precursor, 7103 as complement component 3 precursor, 8006 as testis-specific protein Y, 8101 as Transferrin, 9001 as(Alpha-Oxy, Beta-(C112g)deoxy) T-State Human Hemoglobin, and 9003 as human hemoglobin. 4. Immune protein CR2-C3d, which acts against microbes and pathogenic organisms, and Antitrypsin(803), which is secreted with inflammatory response in the lungs, were increased by more than 200% after the administration of herbal acupuncture. 5. Immunoglobulin lambda chain(3105), Alpha-Oxy, Beta-(C112g)deoxy T-State Human Hemoglobin(9001), and human hemoglobin(9003) were increased by more than two-times after the administration of herbal acupuncture. 6. Proapolipoprotein(2013, 3010) and apolipoprotein(7104), key components of the HDL-cholesterol which plays an important role in preventing arteriosclerosis, were increased after the administration of herbal acupuncture. 7. Vitamin D binding protein(DBP, 2403), protecting the lung at the time of inflammatory response, was increased after the administration of herbal acupuncture. 8. Transthyretin(TTR, 3205), which is the main protein causing familial aimyloid polyneuropathy(FAP), was decreased after the administration of herbal acupuncture. 9. Ras-related protein Ral-A(4002) that controls phospholipid metabolism, cytoskeletal formation, and membrane traffic, was increased after the administration of herbal acupuncture. 10. Testis-specific protein Y(8006), which takes part in determination of the gender, was increased by more than two-times after the administration of herbal acupuncture. 11. Transferrin(8101), T-State Human Hemoblobin(9001), and Human Hemoblobin(9003) which balances the iron level in the body, were increased after the administration of herbal acupuncture. Conousion : Above results support the notion that intravenous injection of cultivated wild ginseng herbal acupuncture induce changes in serum proteins and this research can be a pioneer work in finding biomarkers.

• Parasites, Hosts and Diseases
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• v.54 no.1
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• pp.39-46
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• 2016

Theileria annulata is a tick-borne intracellular protozoan parasite that causes tropical theileriosis, a fatal bovine lymphoproliferative disease. The parasite predominantly invades bovine B lymphocytes and macrophages and induces host cell transformation by a mechanism that is not fully comprehended. Analysis of signaling pathways by quantitative real-time PCR (qPCR) could be a highly efficient means to understand this transformation mechanism. However, accurate analysis of qPCR data relies on selection of appropriate reference genes for normalization, yet few papers on T. annulata contain evidence of reference gene validation. We therefore used the geNorm and NormFinder programs to evaluate the stability of 5 candidate reference genes; 18S rRNA, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ACTB (${\beta}-actin$), PRKG1 (protein kinase cGMP-dependent, type I) and TATA box binding protein (TBP). The results showed that 18S rRNA was the reference gene most stably expressed in bovine PBMCs transformed and non-transformed with T. annulata, followed by GAPDH and TBP. While 18S rRNA and GAPDH were the best combination, these 2 genes were chosen as references to study signaling pathways involved in the transformation mechanism of T. annulata.

• Journal of the East Asian Society of Dietary Life
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• v.7 no.4
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• pp.484-490
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• 1997

In an attempt to investigate the effects of white ginseng on the proliferation and the nitric oxide(NO) secretion of mouse peritoneal macrophages, which are the first mai or defense phagocytes in the immune system, the studies have been carried out. In the macrophage proliferation assay using the $^3$H-thymidine incorporation, the total saponin or Ginsenoside Rb$_2$ were added to the medium at the concentration of 0 to 256$\mu\textrm{g}$/$m\ell$. DNA synthesis of the macrophage was increased at 64$\mu\textrm{g}$/$m\ell$ of total saponin and either 16$\mu\textrm{g}$/$m\ell$ or 64$\mu\textrm{g}$/$m\ell$ of Ginsenoside Rb$_2$, respectively. Also, the effect o(white ginseng on the nitric oxide secretion of the macrophages was investigated. The addition of either total saponin or Ginsenoside Rb$_2$ at the concentration of 20$\mu\textrm{g}$/$m\ell$ significantly increased the secretion of NO from the macrophages. The nitric oxide synthase (NOS) gene expression which is responsible for the synthesis of the nitric oxide has been studied using reverse transcription polymerase chain reaction. In RT-PCR, the $\beta$-actin and nos gene expression have been analyzed. 20$\mu\textrm{g}$/$m\ell$ of either total saponin or Ginsenoside Rb$_2$ increased nos gene expression of the macrophages.

• Journal of Periodontal and Implant Science
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• v.32 no.2
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• pp.389-402
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• 2002

BMP can induce ectopic bone formation when implanted into sites such as rat muscle and can greatly enhance healing of bony defects when applied exogenously. In addition, BMP stimulated osteoblastic differentiation in vitro in various types of cells. The aim of this study was to investigate the effect of recombinant human bone morphogenetic protein(rhBMP-2) on the proliferation and osteoblastic differentiation of human periodontal ligament cells and gingival fibroblasts. The cell number and alkaline phosphatase activity were measured in 3 experimental groups of human periodontal ligament cells and gingival fibroblasts (control group, rhBMP-2 50ng/ml group, and rhBMP-2 100ng/ml group) at 1 and 2 weeks after culture. At the same time, total RNA of cultured cells were extracted and reverse trascription polymerase chain reaction(RT-PCR) was performed to determine the expression of mRNA of bone matrix protein. RhBMP-2 had no effect on the cell proliferation of human periodontal ligament cells and gingival fibroblasts. Alkaline phosphatase activity was elevated significantly by rhBMP-2 in both cells. And periodontal ligament cells showed significantly higher alkaline phosphatase activity than gingival fibroblasts. ${\beta}$-actin, type I collagen, alkaline phosphatase, BMP-2 mRNA were expressed in all of the samples. Osteopontin, osteocalcin mRNA were expressed in all periodontal ligament cell groups, and rhBMP-2 50ng/ml group, rhBMP-2 100ng/ml group of 2 week culture period of gingival fibroblasts. Bone sialoprotein mRNA was only expressed in rhBMP-2 50ng/ml group and rhBMP-2 100ng/ml group of 2-week culture period. These results suggest that rhBMP-2 stimulates osteoblastic differentiation in human periodontal ligament cells and gingival fibroblasts in vitro.

• Journal of Pharmacopuncture
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• v.9 no.3 s.21
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• pp.23-35
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• 2006

Objectives : The aim of this study was to investigate the effect of Asthma-depression and Immunoregulation with PR-HAS(Herbal-acupuncture with Platycodi Radix infusion solution) infection at Joksamni(St36) on ovalbumin-induced asthma in mice. Methods : C57BL/6 mice were sensitized and challenged with OVA(ovalbumin) for 12 weeks(once a week). The experimental group(OVA-PR-HA) was treated with concentrations(1%) of PR-HAS at Joksamni(St36) for the later 8 weeks(3times/week). The second experimental group(OVA-Needle prick) was treated with Needle-Prick at Joksamni(St36) for the later 8 weeks(3times/week). Results : 1. The weight and total cells in the mice lung treated with PR-HA decreased significantly compared with that of control group. 2. Total leukocytes and eosinophils in BALF of the mice group treated with PR-HA decreased remarkably compared with those of control group. 3. The sticking of collagen on histological analysis of lung sections, the mice group treated with PR-HA decreased significantly compared with those of control group. 4. The concentrations of IL-4, IL-5, IgE in BALF, and IL-4, IL-5, IL-13 in serum of the mice group treated with PR-HA decreased significantly compared with those of control group. 5. The number of Gr-1+/CD11b+ and CD11b+ cells in the lungs of the mice group treated with PR-HA decreased significantly compared with that of control group. 6. The numbers of CCR3+ cells, CD4+ cells and CD8+ cells in the lungs, and CD3e+/CD69+ in the lungs of the mice group treated with PA-HA decreased significantly compared with those of control group. 7. The mRNA expression of ${\beta}$-actin, TNF-${\alpha}$, IL-4, IL-5, IL-13 in the mice group treated with PR-HA with RT-PCR decreased significantly compared with those of control group. Conclusions : These result suggests that Platycodi Radix Herbal-acupuncture at Joksamni(St36) in C57BL/6mice may be an effictive part to OVA-induced asthma in C57BL/6 mice.

• Proceedings of the Korea Society of Poultry Science Conference
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• 2004.11a
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• pp.23-25
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• 2004

This study was conducted to investigate the effects of dietary supplementation of Ecklonia cava kjellman(ECK) and crude lectin extracted from ECK (CLEEC) on performances and immune responses in broiler chicks. A total of two hundreds thirty four 1 day old male broiler chicks (Ross) were fed corn-soy based diets containing 0 % (with or without vaccination and Salmonella challenge), 1.0 % ECK, 0.05 %, 0.1 % and 0.3 % CLEEC for 38 days and vaccinated against inactivated ND-IB combined oil vaccine on the fourth day. After S. gallinarum challenge. mortality was measured daily. The spleens of birds were removed for RNA extraction and reverse transcription polymerase chain reaction (RT-PCR) with primer sets for IFN-ν, IL-2, IL-6 and ${\beta}$-actin were performed with RNA samples. At the 28th day, pancreas weights were heavier 0.3 % CLEEC than 1.0 % ECK group. At the 21st day after ND-IB oil vaccine injection, dietary supplementation of ECK and CLEEC tended to increase or significantly (P<0.05) improved ND or IB titer compared to the positive control. Mortality was significantly (P<0.05) decreased by dietary CLEEC treatments. Chicken splenic IFN-ν, IL-2, and IL-6 cytokines mRNA expressions were enhanced by challenge with S, gallinarum. Dietary treatments did not affect mRNA expression of IFN-ν. However, IL-2 and IL-6 expressions in Salmonella challenged birds that fed the 1.0 % ECK or 0.05 % CLEEC groups were enhanced (P<0.05) compare to the positive control. The results demonstrated that dietary ECK and CLEEC enhanced humoral and cellular immunity and therefore. it can be concluded that dietary supplementation of ECK and CLEEC can be used as a feed additive for enhancement of immunocompetence without any adverse effects in broiler chicks.

• The Journal of Internal Korean Medicine
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• v.22 no.4
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• pp.601-611
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• 2001

Objectives : We aimed to identify the dose-dependent inhibitory effects of Cheonsagunja-tang(喘四君子湯), leech(Hirudo medicinalis/水蛭) roasted with Ephedrae Herba(麻黃) on the mRNA expression of IL-6, IL-16, GM-CSF involved in the asthma model. Methods : In the study BEAS-2B cell lines, human epithelial cells were used. These cells were stimulated with tumor necrosis factor(TNF)-${\alpha}$ for artificial inflammatory expression. ${\beta}$-actin messenger RNA(mRNA) was used by internal standard. After 24 hours of the Cheonsagunja-tang, leech-treatment, total cellular RNAs were collected treating RNA zol directly on the living cells. Then the transcriptional activities of IL-6, 16 and GM-CSF were measured by RT-PCR with electrophoresis. Results: In the Cheonsagunja-tang study, the mRNA expression of IL-6 showed 30% transcriptional inhibitory effect compared to the control group in the $100{\mu}l/ml$ category(p<0.005). In the IL-16, there was 26%, 31% and 31% transcriptional inhibitory effect compared to the control groups in the $4{\mu}l/ml$, $20{\mu}l/ml$ and $100{\mu}l/ml$ categories, respectively(p<0.05). In the GM-CSF, the experimental group had 56% transcriptional inhibitory effect compared to the control group in the $100{\mu}l/ml$ category(p<0.001). In other concentrations, there was no inhibitory effect. In the leech study, the mRNA expression of IL-6 showed 37% transcriptional inhibitory effect compared to the control group in the $100{\mu}l/ml$ category(p<0.001). In the IL-16, there was 63% and 67% transcriptional inhibitory effect compared to the control groups in the $20{\mu}/ml$ and $100{\mu}/ml$ categories, respectively(p<0.001). In the GM-CSF, there was 64% and 68% transcriptional inhibitory effect compared to the control groups in the $20{\mu}l/ml$ and $100{\mu}l/ml$ categories, respectively(p<0.001). In other concentrations, there was no inhibitory effect. Conclusions : This study shows that Cheonsagunja-tang and leech roasted with Ephedrae Herba have dose-dependent inhibitory effects on the mRNA expression of IL-6, IL-16 and GM-CSF in BEAS-2B cell lines, human epithelial cells.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.11
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• pp.1649-1656
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• 2015

Gene expression profiling has offered new insights into postmortem molecular changes associated with meat quality. To acquire reliable transcript quantification, high quality RNA is required. The objective of this study was to analyze integrity of RNA isolated from chicken skeletal muscle (pectoralis major) and its capability of serving as the template in quantitative real-time polymerase chain reaction (qPCR) as a function of postmortem intervals representing the end-points of evisceration, carcass chilling and aging stages in chicken abattoirs. Chicken breast muscle was dissected from the carcasses (n = 6) immediately after evisceration, and one-third of each sample was instantly snap-frozen and labeled as 20 min postmortem. The remaining muscle was stored on ice until the next rounds of sample collection (1.5 h and 6 h postmortem). The delayed postmortem duration did not significantly affect $A_{260}/A_{280}$ and $A_{260}/A_{230}$ ($p{\geq}0.05$), suggesting no altered purity of total RNA. Apart from a slight decrease in the 28s:18s ribosomal RNA ratio in 1.5 h samples (p<0.05), the value was not statistically different between 20 min and 6 h samples ($p{\geq}0.05$), indicating intact total RNA up to 6 h. Abundance of reference genes encoding beta-actin (ACTB), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), hypoxanthine-guanine phosphoribosyltransferase (HPRT), peptidylprolylisomerase A (PPIA) and TATA box-binding protein (TBP) as well as meat-quality associated genes (insulin-like growth factor 1 (IGF1), pyruvate dehydrogenase kinase isozyme 4 (PDK4), and peroxisome proliferator-activated receptor delta (PPARD) were investigated using qPCR. Transcript abundances of ACTB, GAPDH, HPRT, and PPIA were significantly different among all postmortem time points (p<0.05). Transcript levels of PDK4 and PPARD were significantly reduced in the 6 h samples (p<0.05). The findings suggest an adverse effect of a prolonged postmortem duration on reliability of transcript quantification in chicken skeletal muscle. For the best RNA quality, chicken skeletal muscle should be immediately collected after evisceration or within 20 min postmortem, and rapidly preserved by deep freezing.

• Tuberculosis and Respiratory Diseases
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• v.42 no.1
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• pp.35-41
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• 1995

Background: Tuberculous cervical lymphadenitis can be diagnosed by clinical findings, chest X-ray, Mantoux test, but confirmed only by excisional biopsy. The polymerase chain reaction(PCR) is now widely applied to test very small amount of pathogen and would be used to detect Mycobacterium tuberculosis in biopsied tissues and fine needle aspirates. Method: We carried out the PCR using IS-1 and IS-2 primers in 16 samples from tuberculous cervical lymphadenitis patients, and 13 samples from non-tuberculous cervical lymphadenopathy patients. Acid fast staining and culture for Mycobacterium were all negative. Results: All of 8 pathologically confirmed tuberculous cervical lymphadenitis samples showed positive PCR results, and of 5/8 clinically diagnosed samples were positive. None of 6 pathologically excluded samples were positive, and among 7 clinically undiagnosed samples 2 showed positive PCR results. Conclusion: In patients with suspected tuberculous cervical lymphadenitis, PCR could be used to detect Mycobacterium tuberculosis using biopsied tissues and even fine needle aspirates with good sensitivity and specificity.

• Journal of Life Science
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• v.28 no.4
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• pp.429-434
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• 2018

In this study, we investigated the potential of Glechoma hederacea var. longituba 70% ethanol extract as a natural functional material by examining the anti-inflammatory effect of it. Macrophages results in (Raw 264.7) induced by lipopolysaccharide (LPS). Confirming the viability of the macrophages Glechoma hederacea var. longituba 70% ethanol extract showed a 95.8% survival rate at $1,000{\mu}g/ml$ concentration. Anti-inflammatory activity was examined the inhibitory tests on the production of LPS included nitric oxide (NO) in RAW 264.7 cells by Griess assay. The result showed that NO production deterrent effect of 37.4% at a concentration of $1,000{\mu}g/ml$. The deterrent effect of GG 70% ethanol extract on protein expression of inducible NOS (iNOS) and Cyclooxygenase-2 (COX-2) was measured by Western blotting using the concentrations 50, 100 and $500{\mu}g/ml$, with ${\beta}$-actin used as the positive control. The inhibitory effect of iNOS and COX-2 mRNA expression was measured by reverse transcription-polymerase chain reaction (RT-PCR) using 50, 100 and $500{\mu}g/ml$ concentration of GG 70% ethanol extract, with GAPDH used as the positive control. In experiments using Western blot and RT-PCR when compared with the control group vitamin C it was confirmed that the 70% ethanol extract from GG suppressed. When compiling the results of this study, we confirmed the possibility of GG 70% ethanol extract as an anti-inflammatory material.