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Effects of Folium Artemisiae Argyi' Herbal Acupuncture of $LR_3$ on Transient Forebrain Ischemic Injury in Rats (태충(太衝) 애엽(艾葉) 약침(藥鍼)이 일과성(一過性) 전뇌(前腦) 허혈(虛血) 손상(損傷)에 미치는 효과(效果))

  • Jang, Jin-Yo;Kim, Jae-Hyo;Park, Sung-Seup;Park, Gyi-Jong;Kim, Kyung-Sik;Sohn, In-Chul
    • Korean Journal of Acupuncture
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    • v.22 no.3
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    • pp.63-81
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    • 2005
  • Herbal acupuncture has been used to prevent and treat the cerebrovascular accident, such as a stroke, and many studies of acupuncture and moxibustion concerning to the stroke have been undertaken in the human and various animals. The main characteristics of herbal acupuncture are a combination of acupuncture and herbal medicine. It was not well known the therapeutic effect and the mechanism of herbal acupuncture on transient forebrain ischemic injury, although it has been used frequently in clinics. In this study, neuroprotective effects of folium artemisiae argyi (艾葉)' herbal acupuncture on the $LR_3$ (Taechung) on the transient forebrain ischemia were investigated in Sprague-Dawely rats. Expressions of cFos, FosB and BDNF protein in the hippocampus and cortex were observed at 2 hrs and 48 hrs after transient forebrain ischemia by immunohistochemistry and ELISA technique. Expression of cFos protein was increased slightly in the hippocampus and cortex at 2 hrs after transient forebrain ischemia, but FosB protein was increased highly comparing to cFos protein. However, pretreatment with folium artemisiae argyi' herbal acupuncture on $LR_3$ significantly increased expression of cFos protein and significantly decreased expression of FosB protein compared to control group. These features were observed in the motor cortex and retrosplenial granular cortex as well as the hippocampus. Also, pretreatment with folium artemisiae argyi' herbal acupuncture on $LR_3$ significantly increased the expression of BDNF protein in the hippocampus ($263.26{\pm}44.56pg/ml$ extracted by water, $275.35{\pm}51.47pg/ml$ extracted by moxa tar)and the cortex ($102.33{\pm}13.65pg/ml$ extracted by water, $109.54{\pm}9.37pg/ml$ extracted by moxa tar) compared to the hippocampus $(134.07{\pm}2.96pg/ml)$ and the cortex $(61.16{\pm}4.11pg/ml)$ in control group at 48 hrs after transient forebrain ischemia. These results suggest that pretreatment with folium artemisiae argyi'herbal acupuncture on $LR_3$ has neuroprotective effect on transient forebrain ischemia and the herbal acupunture on $LR_3$ may be related to antioxidative function of folium artemisiae argyi.

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Saponins from Panax japonicus ameliorate age-related renal fibrosis by inhibition of inflammation mediated by NF-κB and TGF-β1/Smad signaling and suppression of oxidative stress via activation of Nrf2-ARE signaling

  • Gao, Yan;Yuan, Ding;Gai, Liyue;Wu, Xuelian;Shi, Yue;He, Yumin;Liu, Chaoqi;Zhang, Changcheng;Zhou, Gang;Yuan, Chengfu
    • Journal of Ginseng Research
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    • v.45 no.3
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    • pp.408-419
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    • 2021
  • Background: The decreased renal function is known to be associated with biological aging, of which the main pathological features are chronic inflammation and renal interstitial fibrosis. In previous studies, we reported that total saponins from Panax japonicus (SPJs) can availably protect acute myocardial ischemia. We proposed that SPJs might have similar protective effects for aging-associated renal interstitial fibrosis. Thus, in the present study, we evaluated the overall effect of SPJs on renal fibrosis. Methods: Sprague-Dawley (SD) aging rats were given SPJs by gavage beginning from 18 months old, at 10 mg/kg/d and 60 mg/kg/d, up to 24 months old. After the experiment, changes in morphology, function and fibrosis of their kidneys were detected. The levels of serum uric acid (UA), β2-microglobulin (β2-MG) and cystatin C (Cys C) were assayed with ELISA kits. The levels of extracellular matrix (ECM), matrix metalloproteinases (MMPs), tissue inhibitors of metalloproteinases (TIMPs), inflammatory factors and changes of oxidative stress parameters were examined. Results: After SPJs treatment, SD rats showed significantly histopathological changes in kidneys accompanied by decreased renal fibrosis and increased renal function; As compared with those in 3-month group, the levels of serum UA, Cys C and β2-MG in 24-month group were significantly increased (p < 0.05). Compared with those in the 24-month group, the levels of serum UA, Cys C and β2-MG in the SPJ-H group were significantly decreased. While ECM was reduced and the levels of MMP-2 and MMP-9 were increased, the levels of TIMP-1, TIMP-2 and transforming growth factor-β1 (TGF-β1)/Smad signaling were decreased; the expression level of phosphorylated nuclear factor kappa-B (NF-κB) was down-regulated with reduced inflammatory factors; meanwhile, the expression of nuclear factor erythroid 2-related factor 2-antioxidant response element (Nrf2-ARE) signaling was aggrandized. Conclusion: These results suggest that SPJs treatment can improve age-associated renal fibrosis by inhibiting TGF-β1/Smad, NFκB signaling pathways and activating Nrf2-ARE signaling pathways and that SPJs can be a potentially valuable anti-renal fibrosis drug.

Anti-allergic Effects of Gagam-YangGyeokSan on RBL-2H3 Mast Cells and OVA/alum Sensitized Mice (가감양격산(加減凉膈散)이 RBL-2H3 비만세포와 OVA/alum에 감작된 생쥐에 미치는 항알레르기 효과)

  • Lee, Yun Shil;Han, Jae Kyung;Kim, Yun Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.26 no.4
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    • pp.10-23
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    • 2012
  • Objectives: Gagamyanggyeoksan (G-YGS) has been used to suppress allergic reaction, however, the cellular target of G-YGS and its mode of action remain unclear. The present study was designed to investigate the effect of extracted G-YGS on the PMA and lonomycin (PI)-induced activation of RBL-2H3. Methods: For this investigation, We examined IL-4, IL-13 mRNA expression by Real-Time PCR, IL-4, IL-13 production by ELISA analysis and manifestations of GATA-1, GATA-2, NF-AT1, NF-AT2, AP-1 and NF-${\kappa}B$ p65 transcription factors by western blotting, OVA-specific IgE, IL-4, IL-13 by mouse be sensitive to OVA. Results: Here we showed that treatment of RBL-2H3 mast cells with G-YGS, suppressed PI-induced production of Th2 cytokines including IL-4 and IL-13 in a dose dependent manner. The mRNA expression of IL-4 were completely abolished by G-YGS at the concentration of $100{\mu}g/ml$. Data from a stable cell lines consistently expressing IL-4. And the mRNA expression of IL-13 were abolished by G-YGS at the $200{\mu}g/ml$. But there is no difference between the $50{\mu}g/ml$, the $100{\mu}g/ml$ and the comparison. Results from the western blot analysis of transcription factors involving IL-4 and IL-13 expression indicated that it prominently decreased the expression of mast cell specific transcricption factors including GATA-1, GATA-2, NF-AT2, c-Jun, NF-${\kappa}B$ p65 but not c-Fos. And G-YGS suppressed IgE, IL-4, IL-13 in mouse be sensitive to OVA. Conclusions We suggested the anti-allergic activities of G-YGS might be mediated by down-regulation of Th2 cytokines such as IL-4 and IL-13 through the regulation of transcription factors as GATA-1, GATA-2, NF-AT2, c-Jun, NF-${\kappa}B$ p65.

Assessment of Validity of RT-PCR and EIA for The Detection of Hepatitis C Virus Infection (C형 간염의 진단을 위한 역전사중합효소연쇄반응과 효소면역측정법의 타당성 평가)

  • Son, Byung-Chul;Chun, Jin-Ho;Park, Yeong-Hong;Shin, Hai-Rim;Cho, Kyu-Il;Kim, Jong-Han;Jung, Kui-Oak;Lee, Jong-Tae;Lee, Chae-Un;Paik, Nak-Whan
    • Journal of Preventive Medicine and Public Health
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    • v.28 no.2 s.50
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    • pp.526-541
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    • 1995
  • This study was conducted to estimate the validity of reverse transcriptase-polymerase chain reaction(RT-PCR) compared to enzyme immunoassay(EIA) for the detection of hepatitis C virus (HCV) infection. EIA for antibody to HCV(anti-HCV) and RT-PCR for HCV was executed on the subjects from Pusan and Kyungnam area with questionnaire survey to collect some relating factors of HCV infection. As the result from 617 cases, the prevalence of HCV infection was 1.5% by EIA and 3.7% by RT-PCR(p<0.05), and the age standardized rate was 1.7% and 3.4% by EIA and RT-PCR, respectively. The prevalence of hepatitis B surface antigen(HBsAg) was 6.8% by enzyme linked immunosorbent assay(ELISA) and the age standardized rate was 7.7%. It was the higher in male group comparing to female group(p<0.01). Both of the prevalence of HCV and HBsAg were higher in elevated asparate aminotransferase(AST) and alanine aminotransferase (ALT) group than in normal AST and ALT group(p<0.01). There was no specific risk factor of HCV infection. Though the degree of agreement of EIA and RT-PCR by gamma statistics was 97.2%, it showed a significant difference between the two methods(p<0.01). For the detection of HCV infection, positive predictive value of EIA was 66.7% and negative predictive value of EIA was 97.2%. This study suggests that negative result to anti-HCV by EIA didn't mean the free state of HCV infection, therefore it would be helpful that further monitoring for HCV infection by RT-PCR in the case of elevated AST and ALT and/or clinically suspected.

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Regulation of Chilling Tolerance in Rice Seedlings by Plant Hormones

  • Chu, Chun;Lee, Tse-Min
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.37 no.3
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    • pp.288-298
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    • 1992
  • Since the major important factors limiting plant growth and crop productivity are environmental stresses, of which low temperature is the most serious. It has been well known that many physiological processes are alterant in response to the environmental stress. With regard to the relationship between plant hormones and the regulation of chilling tolerance in rice seedlings, the major physiological roles of plant hormones: abscisic acid, ethylene and polyamines are evaluated and discussed in this paper. Rice seedlings were grown in culture solution to examine the effect of such plant hormones on physiological characters related to chilling tolerance and also to compare the different responses among tested cultivars. Intact seedlings about 14 day-old were chilled at conditions of 5$^{\circ}C$ and 80% relative humidity for various period. Cis-(+)-ABA content was measured by the indirect ELISA technique. Polyamine content and ethylene production in leaves were determined by means of HPLC and GC respectively. Chilling damage of seedlings was evaluated by electrolyte leakage, TTC viability assay or servival test. Our experiment results described here demonstrated the physiological functions of ABA, ethylene, and polyamines related to the regulation of chilling tolerance in rice seedlings. Levels of cis-(+)-ABA in leaves or xylem sap of rice seedlings increased rapidly in response to 5$^{\circ}C$ treatment. The tolerant cultivars had significant higher level of endogenous ABA than the sensitive ones. The ($\pm$)-ABA pretreatment for 48 h increased the chilling tolerance of the sensitive indica cultivar. One possible function of abscisic acid is the adjustment of plants to avoid chilling-induced water stress. Accumulation of proline and other compatible solutes is assumed to be another factor in the prevention of chilling injuies by abscisic acid. In addition, the expression of ABA-responsive gene is reported in some plants and may be involving in the acclimation to low temperature. Ethylene and its immediate precusor, 1-amincyclopropane-1-carboxylic acid(ACC) increased significantly after 5$^{\circ}C$ treatment. The activity of ACC synthase which converts S-adenosylmethionine (SAM) to ACC enhanced earlier than the increase of ethylene and ACC. Low temperature increased ACC synthase activity, whereas prolonged chilling treatment damaged the conversion of ACC to ethylene. It was shown that application of Ethphon was beneficial to recovering from chilling injury in rice seedlings. However, the physiological functions of chilling-induced ethylene are still unclear. Polyamines are thought to be a potential plant hormone and may be involving in the regulation of chilling response. Results indicated that chilling treatment induced a remarkable increase of polyamines, especially putrescine content in rice seedlings. The relative higher putrescine content was found in chilling-tolerant cultivar and the maximal level of enhanced putrescine in shoot of chilling cultivar(TNG. 67) was about 8 folds of controls at two days after chilling. The accumulation of polyamines may protect membrane structure or buffer ionic imbalance from chilling damage. Stress physiology is a rapidly expanding field. Plant growth regulators that improve tolerance to low temperature may affect stress protein production. The molecular or gene approaches will help us to elucidate the functions of plant hormones related to the regulation of chilling tolerance in plants in the near future.

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Characterization of Humanized Antibody Produced by Apoptosis-Resistant CHO Cells under Sodium Butyrate-Induced Condition

  • Kim, No-Soo;Chang, Kern-Hee;Chung, Bo-Sup;Kim, Sung-Hyun;Kim, Jung-Hoe;Lee, Gyun-min
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.926-936
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    • 2003
  • Overexpression of human Bcl-2 protein in recombinant Chinese hamster ovary (rCHO) cells producing humanized antibody (SH2-0.32) considerably suppressed sodium butyrate (NaBu)-induced apoptosis during batch culture by using commercially available serum-free medium, which extended the culture longevity. Due to the extended culture longevity provided by the anti-apoptotic effect of Bcl-2 overexpression, the final antibody concentration of 14C6-bcl-2 culture (Bcl-2 high producer, $23\;\mu\textrm{g}\;ml^{-1}$) was 2 times higher than that of the $SH2-0.32-{\Delta}bcl-2$ culture (cells transfected with bcl-2-deficient plasmid, $10.5\;\mu\textrm{g}\;ml^{-1}$) in the presence of NaBu. To determine the effect of NaBu/Bcl-2 overexpression on the molecular integrity of protein products, antibodies purified from 14C6-bcl-2 and $SH2-0.32-{\Delta}bcl-2$ cultures in the presence of NaBu were characterized by using various molecular assay systems. For comparison, antibody purified from the parental rCHO cell culture (SH2-0.32) in the absence of NaBu was also characterized. No significant changes in molecular weight of antibodies could be observed by SDS-PAGE. From GlycoSep-N column analysis, it was found that the core oligosaccharide structure ($GlcNAc_2Man_3GlcNAc_2$) was not affected by NaBu/Bcl-2 overexpression, while the microheterogeneity of N-linked oligosaccharide structure was slightly affected. Compared with the antibody produced in the absence of NaBu, the proportion of neutral oligosaccharides was increased from 10% (14C6-bcl-2) to 16% ($SH2-0.32-{\Delta}bcl-2$) in the presence of NaBu, which was accompanied by the reduced proportion of acidic oligosaccharides, especially of monosialylated and disialylated forms. The changes in microheterogeneous oligoformal structures of antibody in turn affected the mobility of antibody isoforms in isoelectric focusing (IEF), resulting in the occurrence of some more basic antibody isoforms produced in the presence of NaBu. However, the antigen-antibody binding properties were not changed by alteration of glycosylation pattern. The competitive enzyme-linked immunosorbent assay (ELISA) showed that the antibody produced by NaBu/Bcl-2 overexpression maintained its antigen-antibody binding properties with binding affinity of about $2.5{\times}10^9{\;}M^{-1}$. Taken together, no significant effects of NaBu/Bcl-2 overexpression on the molecular integrity of antibodies, produced by using serum-free medium, could be observed by the molecular assay systems.

The Suppressive Effect of Th2 Cytokines Expression and the Signal Transduction Mechanism in MC/9 Mast Cells by Forsythiae Fructus Extracts (비만세포에서 연교(連翹) 추출물의 Th2 사이토카인 발현 및 신호전달 기전 억제 효과)

  • Lee, Jin Hwa;Han, Jae Kyung;Kim, Yun Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.28 no.3
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    • pp.31-46
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    • 2014
  • Objectives Forsythiae Fructus treatment has been used for inflammatory and allergic diseases in Korean Medicine. Nevertheless, the mechanism of action and the cellular targets are not understood well. The pathogenesis of allergic diseases are associated with Th2 cytokines such as IL-13, MIP-$1{\alpha}$, IL-13, IL-5, GM-CSF, IL-4, TNF-${\alpha}$ and IL-6, which are secreted by the mast cells. This study was conducted to investigate the effects of Forsythiae Fructus extracts (FF) on Th2 cytokines expression and signal transduction in MC/9 mast cells. Methods In the study, MC/9 mast cells were stimulated with DNP-IgE for 24 hours and then treated separately with CsA $10{\mu}g/m{\ell}$ and varying doses of FF for one hour. MC/9 mast cells stimulated with DNP-IgE was the control group, a treatment with CsA was the positive control group and a treatment with varying doses FF was the experimental groups. The mRNA levels of IL-13, IL-5, GM-CSF, IL-4, TNF-${\alpha}$, IL-6 were analyzed with Real-time PCR. The levels of IL-13, MIP-$1{\alpha}$ were measured using enzyme-linked immunosorbent assays(ELISA). NFAT, AP-1 and NF-${\kappa}B$ p65 were examined by Western blot analysis. Results 1. FF were observed to suppress the mRNA expression of IL-13, IL-5, GM-CSF, IL-4, TNF-${\alpha}$, IL-6 in comparison to DNP-IgE control group. 2. FF also has inhibited the IL-13, MIP-$1{\alpha}$ production significantly in comparison to DNP-IgE control group. 3. Western blot analysis of transduction factors involving Th2 cytokines expression has revealed a prominent decrease of the mast cell specific transduction factors including NFAT-1, NFAT-2, c-Jun, and NF-${\kappa}B$ p65 but c-Fos. Conclusions In conclusion, the anti-allergenic activities of FF may be strongly related to the regulation of transcription factors NFAT-1, NFAT-2, c-Jun, and NF-${\kappa}B$ p65 causing inhibition of Th2 cytokines in mast cells.

Effects of natural raw meal (NRM) on high-fat diet and dextran sulfate sodium (DSS)-induced ulcerative colitis in C57BL/6J mice

  • Shin, Sung-Ho;Song, Jia-Le;Park, Myoung-Gyu;Park, Mi-Hyun;Hwang, Sung-Joo;Park, Kun-Young
    • Nutrition Research and Practice
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    • v.9 no.6
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    • pp.619-627
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    • 2015
  • BACKGROUND/OBJECTIVES: Colitis is a serious health problem, and chronic obesity is associated with the progression of colitis. The aim of this study was to determine the effects of natural raw meal (NRM) on high-fat diet (HFD, 45%) and dextran sulfate sodium (DSS, 2% w/v)-induced colitis in C57BL/6J mice. MATERIALS/METHODS: Body weight, colon length, and colon weight-to-length ratio, were measured directly. Serum levels of obesity-related biomarkers, triglyceride (TG), total cholesterol (TC), low density lipoprotein (LDL), high density lipoprotein (HDL), insulin, leptin, and adiponectin were determined using commercial kits. Serum levels of pro-inflammatory cytokines including tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin (IL)-$1{\beta}$, and IL-6 were detected using a commercial ELISA kit. Histological study was performed using a hematoxylin and eosin (H&E) staining assay. Colonic mRNA expressions of TNF-${\alpha}$, IL-$1{\beta}$, IL-6, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) were determined by RT-PCR assay. RESULTS: Body weight and obesity-related biomarkers (TG, TC, LDL, HDL, insulin, leptin, and adiponectin) were regulated and obesity was prevented in NRM treated mice. NRM significantly suppressed colon shortening and reduced colon weight-to-length ratio in HFD+DSS induced colitis in C57BL/6J mice (P < 0.05). Histological observations suggested that NRM reduced edema, mucosal damage, and the loss of crypts induced by HFD and DSS. In addition, NRM decreased the serum levels of pro-inflammatory cytokines, TNF-${\alpha}$, IL-$1{\beta}$, and IL-6 and inhibited the mRNA expressions of these cytokines, and iNOS and COX-2 in colon mucosa (P < 0.05). CONCLUSION: The results suggest that NRM has an anti-inflammatory effect against HFD and DSS-induced colitis in mice, and that these effects are due to the amelioration of HFD and/or DSS-induced inflammatory reactions.

Luteinizing hormone beta gene polymorphism and its effect on semen quality traits and luteinizing hormone concentrations in Murrah buffalo bulls

  • Reen, Jagish Kour;Kerekoppa, Ramesha;Deginal, Revanasiddu;Ahirwar, Maneesh Kumar;Kannegundla, Uday;Chandra, Satish;Palat, Divya;Das, Dayal Nitai;Kataktalware, Mukund Amritrao;Jeyakumar, Sakthivel;Isloor, Shri krishna
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.8
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    • pp.1119-1126
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    • 2018
  • Objective: Present investigation was aimed to study the Single Nucleotide Variants of the luteinizing hormone beta ($LH{\beta}$) gene and to analyze their association with the semen quality (fresh and post-thawed frozen semen) and luteinizing hormone (LH) concentrations in Murrah buffalo bulls. Methods: Polymerase chain reaction-single stranded conformational polymorphism (PCR-SSCP) and Sanger sequencing method is used to study genetic variability in $LH{\beta}$ gene. LH assay was carried out using enzyme-linked immunosorbent assay method. A fixed general linear model was used to analyze association of single nucleotide polymorphism (SNP) of $LH{\beta}$ gene with semen quality in 109 and LH concentrations in 80 Murrah bulls. Results: $LH{\beta}$ gene was found to be polymorphic. Total six SNPs were identified in $LH{\beta}$ gene g C356090A, g C356113T, g A356701G, g G355869A, g G356330C, and g G356606T. Single Stranded Conformational Polymorphism variants of pattern 2 of exon 1+pattern 2 of exon 2+pattern 1 of exon 3 had highly significant (p<0.01) effect on sperm concentration (million/mL), percent mass motility, acrosome integrity and membrane integrity in fresh and frozen semen whereas significant (p<0.05) effect was observed on percent live spermatozoa. SSCP variants of pattern 2 of exon 1+pattern 2 of exon 2+pattern 1 of exon 3 had highly significant (p<0.01) effect on luteinizing hormone concentrations too. Conclusion: The observed association between SSCP variants of $LH{\beta}$ gene with semen quality parameters and LH concentrations indicated the possibilities of using $LH{\beta}$ as a candidate gene for identification of markers for semen quality traits and LH concentrations in Murrah buffaloes.

Effects of Schizonepeta Spica water extrect on the OVA-induced BALB/c mice model of allergic rhinitis (난알부민으로 알레르기 비염이 유발된 마우스에서 형개의 개선효과)

  • Song, Dea-Uk;Heo, Jun;Lee, SungYub;Kim, Jae-Hyun;Lee, Bina;Min, Ju-Hee;Kim, Eun-Young;Sohn, Youngjoo;Jung, Hyuk-Sang
    • The Korea Journal of Herbology
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    • v.30 no.6
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    • pp.39-46
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    • 2015
  • Objectives : The aim of this study was to evaluate the effect of Schizonepeta Spica water extract (SS) on the OVA-induced BLAB/c mice allergic rhinitis model.Methods : Thirty two BALB/c mice were equally assigned to 4 groups; the sham group, the control group, the cetirizine hydrochloride (Cet) treatment group, and the SS treatment group. Sham group was sensitized and challenged with saline, and the other groups were sensitized and challenged with OVA. The dosage of SS was 7.6 mg /kg·day, and Cet was 10 mg/kg·day. Nasal rubbing and sneezing were measured by the behavior observation. The concentrations of IL-1β, IL-10, TNF-α and MIP-2 in the sera of allergic rhinitis model were measured by mouse cytokine/chemokine magnetic bead panel kits. Total IgE and OVA-specific IgE were measured by ELISA method. Epithelial thickness and eosinophil infiltration of nasal septum was investigated by histological examination.Results : The clinical symptoms that increased in control group were significantly reduced in SS-treated group. Serum total IgE and OVA-specific IgE in the SS-treated group were significantly reduced compared to the control group. The concentration of IL-1β, IL-10, TNF-α and MIP-2 in SS-treated group showed a significant reduction compared to the control group. The infiltration of eosinophil into nasal tissues of SS-treated group decreased markedly compared to control group, and thickness of nasal septum in nasal mucosa showed a significant reduction compared to control group.Conclusions : According to the above result, it is suggested that SS may inhibit the early and late phase of allergic rhinitis reaction.