• 제목/요약/키워드: ELISA method

검색결과 596건 처리시간 0.025초

Effects of Sophora Subprostrata against Focal Cerebral Ischemic Damage by Middle Cerebral Artery Occlusion in Rats (광두근이 백서 중대뇌동맥 폐쇄에 의한 국소뇌허혈손상에 미치는 효과)

  • 이현삼;정혁상;강철훈;손낙원
    • The Journal of Korean Medicine
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    • 제21권2호
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    • pp.68-78
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    • 2000
  • Objective : This research was performed to investigate protective effects of Sophora subprostrata, against ischemic brain damage after a middle cerebral artery(MCA) occlusion. The effect was estimated using histological test, neurobehavioural test, and biochemical test. Methods : Rats(Sprague-Dawley) were divided into four groups: Sham operated group, MCA occluded group, Sophora subprostrata administrated group after MCA occlusion, and Normal group. The MCA was occluded by intraluminal method. Sophora subprostrata was administrated orally twice(l and 4 hours) after middle cerebral artery occlusion. The neurobeavioural test was performed at 3 hours, 6 hours, 9 hours and 24 hours after the surgery by posture reflex test and swimming behavioural test. All groups were sacrificed at 24 hours after the surgery. The brain tissue was stained with 2% triphenyl tetrazolium chioride(TTC) or 1 % cresyl violet solution, to examine effect of Sophora subprostrata on ischemic brain tissue. The blood samples were obtained from the heart of rats. Tumor necrosis factor-a level was measured from sera using Enzyme-Linked Immunoabsorbent Assay(ELISA). Results : The results showed that (1) Sophora subprostrata reduced infarct size and total infarct volume by 54.8% compared to the control group, (2) that neuronal death, which was shown by decrease in cell number and size, was attenuated significantly in the boundary area of the infarction, (3) that serum $TNF-{\alpha}$ㆍlevel was reduced significantly, and finally, there was significant recovery of motor deficit at 3 hours after MCA occluded by Swimming behavioural test. Conclusions :In conclusion, Sophora subprostrata has protective effects against ischemic brain damage at the early stage of ischemia.

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A Clinical Study for Rota Virus Infection of Neonates in Oriental Medicine Hospital Postpartum Care Center (한방병원 산후조리센터 내 신생아의 Rota Virus 감염증에 대한 임상적 고찰)

  • Lee, Jeong-Lim;Ha, Kwang-Su;Ha, Su-Yun;Song, In-Sun;Kim, Sun-Mi
    • The Journal of Pediatrics of Korean Medicine
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    • 제19권2호
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    • pp.115-126
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    • 2005
  • Objective : The aim of this study is to investigate rota virus infection of neonates In the Postpartum care center. Methods : This clinical study was carried out with the 205 neonates who admitted to the Postpartum Care Center of ${\bigcirc}{\bigcirc}$ Oriental Medicine Hospital, from July, 2004 to July, 2005. Stools were examined daily with Rotazyme ELISA tests(Bio Line Rota Virus, SD Standard Diagnostics, INC.) for 3 days from the day of admission. Supplementary test was examined when change was found in stools. Results : Among 205 newborn babies, 58 cases turned out to be positive. The positive rate of rota virus was not significantly related to sex, birth weight, gestation age. But, it was significantly related to delivery method and it was higher in caesarean section group. The manifestation rates of symptoms were 51.7% in Rotazyme positive neonates. In the Rotazyme positive 58 cases, 30 cases(51.7%) had diarrhea, 8 cases had fever(13.8%), 1 case(1.7%) had vomiting. The positive rate of rota virus was higher during from october to march. There were 190 cases(92.2%) of mixed feeding and 15 cases(7.8%) of artificial feeding. There was significance between breast feeding and the positive rate of rota virus. There was no significance between hospital of birth and the rate of positive rotazyme test. Conclusions : A further study of neonates in postpartum care center is necessary as preference of postpartum care center was increased. And the oriental medicine treatment of rota virus IS expected to be effective, so a further study is necessary.

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Hapten Synthesis and Influence of Coating Ligands on Enzyme-linked Immunoreaction of DDT

  • Hong, Ji- Youn;Kim, Jong-Hyun;Choi, Myung-Ja
    • Bulletin of the Korean Chemical Society
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    • 제23권10호
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    • pp.1413-1431
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    • 2002
  • For the development of immunodetection method of 4,4'-dichlorodipheny-2,2,2-trichloroethane (p,p'-DDT), a persistent and broad toxic organochlorine insecticide, various DDT derivatives were synthesized and characterized for the use of immunogens and the coating ligands for the antibody evaluation. The appropriate lengths of linkers were introduced to investigate more efficient DDT derivatives. Among these hapten derivatives, 2,2-Bis(4-chlorophenyl)acetic acid (DDA), 5,5-Bis(4-chlorophenyl)-5-hydroxypentanoic acid (DDHP) and 5,5-Bis(4-chlorophenyl)-5-chloropentanoic acid (DDCP) were conjugated with keyhole limpet hemocyanin (KLH) for the use of immunogen to produce antibodies. 6,6-Bis(4-chlorophenyl)-6-hydroxyhexanoic acid (DDHH) and 3-[6,6-Bis(4-chlorophenyl)-6-hydroxyhexanoylamino]propanoic acid (DDHHAP) in addition to above hapten derivatives were conjugated to ovualbumin (OVA) and bovine serum albumin (BSA) for the use of coating ligands to measure the titration level of antibody and the displacement of free analytes. Three matching pairs of antibodies and coating ligands were selected for the simultaneous detection of p,p'-DDT and its related compounds of DDA and 2,2-bis(4-chlorophenyl)-1,1-dichloroethylene (p,p'-DDE) by investingating the displacement of free analytes in an indirect ELISA. These were PAb #1 and coating ligand DDCP-OVA, PAb #1 and DDHHAP-OVA, and PAb #3 and DDHHAP-OVA. The most useful immunoreaction for DDT analytes were obtained using PAb #3 and coating ligand DDHHAP-OVA showing 3.4 ng/mL of lower limit of detection. These results indicated that titration level and free analytes displacement were greatly influenced by hapten derivatized and carrier proteins conjugated.

Effects of Achyranthis Japonicae Radix-containing mixture on monosodium iodoacetate-induced osteoarthritis in rats (우슬(牛膝) 등 복합 추출물의 monosodium iodoacetate로 유발한 흰쥐 골관절염에 대한 효과)

  • Kim, Myung-Gyou;Seo, Il-Bok;Leem, Kang-Hyun;Jeong, Taejin;Kim, Jinseok
    • The Korea Journal of Herbology
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    • 제32권1호
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    • pp.83-89
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    • 2017
  • Objectives : The present study was designed to determine the effects of mixture of Achyranthis Japonicae Radix, Scutellariae Radix, and Acanthopanacis Cortex on monosodium iodoacetate (MIA)-induced osteoarthritis in rats. The mixture was composed of Achyranthis Japonicae Radix, Scutellariae Radix, and Acanthopanacis Cortex extracts. Methods : Arthritis was induced by injection of MIA into knee joints of rats. At the end of experiment, gross examination on the articular structures of knee joints were performed. Proteoglycan (PG) contents in articular cartilages were analysed as well. Tumor necrosis $factor-{\alpha}$($TNF-{\alpha}$) and $interleukin-1{\beta}$ ($IL-1{\beta}$) contents in synovial fluids were measured by ELISA method and matrix metalloproteinase 2 (MMP2), MMP9, and tissue inhibitors of metalloproteinase 1 (TIMP1) mRNA were measured by a realtime PCR. Results : The surfaces of the articular cartilage were observed. The severity of osteoarthritis in the treated group were alleviated compared with control group. PG contents in articular cartilages of the treated group were increased compared with control group. $IL-1{\beta}$ contents in synovial fluids of the treated group were significantly decreased compared with control group. MMP2 and MMP9 mRNA contents in articular cartilages were significantly decreased compared with control group and TIMP1 mRNA contents were increased compared with control group. Conclusions : On the basis of these results, we concluded that Achyranthis Japonicae Radix-containing mixture treatment has anti-arthritic effects on the MIA-induced osteoarthritis in rats. And the effects were related with the reduction of $IL-1{\beta}$ in synovial membranes and the consequent reduction of MMP2 and MMP9 expressions.

The effect of 100KHz PWM LED light irradiation on RAT bone-marrow cells (100kHz PWM LED 광조사가 백서 골수세포에 미치는 영향)

  • Cheon, Min-Woo;Kim, Seong-Hwan;Kim, Young-Pyo;Lee, Ho-Sic;Park, Yong-Pil;Yu, Seong-Mi;Lee, Hee-Gap;Kim, Tae-Gon
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 한국전기전자재료학회 2008년도 하계학술대회 논문집 Vol.9
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    • pp.512-513
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    • 2008
  • The study examined what effects 100kHz PWM LED light irradiation causes to bone marrow cells of SD-Rat when LED characterized cheap and safe is used onto the light therapy by replacing the low 1evel laser. We developed the equipment palpating cell proliferation using a high brightness LED. This equipment was fabricated using a micro-controller and a high brightness LED, and designed to enable us to control light irradiation time, intensity, frequency and so on. Especially, to control the light irradiation frequency, FPGA was used, and to control the change of output value, TLC5941 was used. Consequent1y, the current value could be controlled by the change of 1eve1 in Continue Wave(CW) and Pulse Width Modulation(PWM), and the output of a high brightness LED could be controlled stage by stage. MTT assay method was chosen to verify the cell increase of two groups and the effect of irradiation on cell proliferation was examined by measuring 590nm transmittance of ELISA reader. As a result, the cell increase of Rat bone marrow cells was verified in 100kHz PWM LED light irradiation group as compared to non-irradiation group.

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2-Nonadecanone Alleviates Depression through Inflammation Relief in SD Rat

  • Lee, Gil-Hyun;Hyun, Kyung-Yae
    • Biomedical Science Letters
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    • 제24권3호
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    • pp.206-212
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    • 2018
  • Depression is a type of mood disorder characterized by hypochondriasis, decreased appetite, and insomnia. Depression is a disease that affects more than 100 million people worldwide. 2-Nonadecanone (NAC) is a bioactive substance that constitutes Fomes fomentarius, and NAC is expected to have an antidepressant effect. By using the forced swimming test (FST), we investigated the effects of treatment with NAC on immobility subacutely in rats after oral dosing once a day for 2 days. Serum levels of cytokine interleukin-1 beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) were determined by enzyme-linked immunosorbent assay (ELISA). Nuclear factor-2 (Nrf-2) and inducible nitric oxide synthases (iNOS) were analyzed by western blot method. NAC dose-dependently decreased immobility in the FST. NAC dosedependently decreased FST-induced increase of cytokine levels, as manifested by significantly stronger effects on $IL-1{\beta}$ and $TNF-{\alpha}$ levels at higher doses than the lowest dose of NAC. Western blot analysis showed that Nrf-2 was significantly lower in the NAC-treated group than in the disease-induced group. The iNOS results were also significantly lower in the NAC-treated group than in the other groups. Considering FST results, the antidepressant effect of NAC is effective. Considering the results of cytokine and protein expression, this anti-depressant effect may be related to the anti-inflammatory effect. Therefore, it can be said that the anti-inflammatory effect of NAC increases the antidepressant effect in the FST experiment.

The Effect of Ginseng Saponin Fractions on Humoral Immunity of Mouse (생쥐의 체액성 면역에 미치는 인삼사포닌 분획물들의 영향)

  • Park, Han-U;Kim, Se-Chang;Jeong, No-Pal
    • Journal of Ginseng Research
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    • 제12권1호
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    • pp.63-67
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    • 1988
  • To investigate the effect of ginseng saponin fractions (total saponin, diol saponin and triol saponin) on the antibody production and on the recovery of immunosuppression in mouse, chick ${\gamma}$-globulin was used as immunogen and CY(cyclophosphamide) as immunosuppressive drug. The effect of ginseng saponin fractions on the production of total serum protein was investigated also. Circulating antibody was measured with ELISA method. Total saponin, dial saponin and triol saponin resulted 4 times higher titer values compared to control group in the production of antibody but resulted no effect on the recovery of immunosuppression induced by CY. From the above results ginseng saponin fractions are believed to effect on intact immune system and to promote antibody production by helping the cooperations among lymphocytes or the growth of lymphocytes. And the increase of total serum protein has no direct relations with the increase of circulatory antibody.

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Inhibition of COX-2 Activity and Proinflammatory Cytokines($TNF-{\alpha}{\;}and{\;}IL-1{\beta}$) Production by Water-Soluble Sub-Fractionated Parts from Bee (Apis mellifera) Venom

  • Nam, Kung-Woo;Je, Kang-Hoon;Lee, Jang-Hurn;Han, Ho-Je;Lee, Hye-Jung;Kang, Sung-Kil;Mar, Woongchon
    • Archives of Pharmacal Research
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    • 제26권5호
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    • pp.383-388
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    • 2003
  • Bee venom is used as a traditional medicine for treatment of arthritis. The anti-inflammatory activity of the n-hexane, ethyl acetate, and aqueous partitions from bee venom (Apis mellifera) was studied using cyclooxygenase (COX) activity and pro-inflammatory cytokines (TNF-$\alpha and IL-1\beta$) production, in vitro. COX-2 is involved in the production of prostaglandins that mediate pain and support the inflammatory process. The aqueous partition of bee venom showed strong dose-dependent inhibitory effects on COX-2 activity ($IC_{50} = 13.1 \mu$ g/mL), but did not inhibit COX-1 activity. The aqueous partition was subfractionated into three parts by molecular weight differences, namely, B-F1 (above 20 KDa), B-F2 (between 10 KDa and 20 KDa) and BF-3 (below 10 KDa). B-F2 and B-F3 strongly inhibited COX-2 activity and COX-2 mRNA expression in a dose-dependent manner, without revealing cytotoxic effects. TNF-$\alpha and IL-1\beta$ are potent pro-inflammatory cytokines and are early indicators of the inflammatory process. We also investigated the effects of three subfractions on TNF-$\alpha and IL-1\beta$ production using ELISA method. All three subfractions, B-F1, B-F2 and B-F3, inhibited TNF-$\alpha and IL-1\beta$production. These results suggest the pharmacological activities of bee venom on anti-inflammatory process include the inhibition of COX-2 expression and the blocking of pro-inflammatory cytokines (TNF-$\alpha and IL-1\beta$) production.

Chracterization of THP-1 Cell Death Induced by Porphyromonas gingivalis Infection

  • Song, YuRi;Kim, SeYeon;Park, Mee Hee;Na, Hee Sam;Chung, Jin
    • International Journal of Oral Biology
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    • 제42권1호
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    • pp.17-23
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    • 2017
  • Background: Periodontitis is generally a chronic disorder characterized by the breakdown of tooth-supporting tissues. P. gingivalis, a Gram-negative anaerobic rod, is one of the major pathogens associated with periodontitis. Frequently, P. gingivalis infection leads to cell death. However, the correlation between P. gingivalis-induced cell death and periodontal inflammation remains to be elucidated. Among cell deaths, the death of immune cells appears to play a significant role in inflammatory response. Thus, the aim of this study was to examine P. gingivalis-induced cell death, focusing on autophagy and apoptosis in THP-1 cells. Methods: Human acute monocytic leukemia cell line (THP-1) was used for all experiments. Autophagy induced by P. gingivalis in THP-1 cells was examined by Cyto ID staining. Intracellular autophagic vacuoles were observed by fluorescence microscopy using staining Acridine orange (AO); and 3-methyladenine (3-MA) was used to inhibit autophagy. Total cell death was measured by LDH assay. Cytokine production was measured by an ELISA method. Results: P. gingivalis induced autophagy in an MOI-dependent manner in THP-1 cells, but 3-MA treatment decreased autophagy and increased the apoptotic blebs. P. gingivalis infection did not increase apoptosis compared to the control cells, whereas inhibition of autophagy by 3-MA significantly increased apoptosis in P. gingivalis-infected THP-1 cells. Inhibition of autophagy by 3-MA also increased total cell deaths and inflammatory cytokine production, including $IL-1{\beta}$ and $TNF-{\alpha}$. Conclusion: P. gingivalis induced autophagy in THP-1 cells, but the inhibition of autophagy by 3-MA stimulated apoptosis, leading to increased cell deaths and pro-inflammatory cytokines production. Hence, the modulation of cell deaths may provide a mechanism to fight against invading microorganisms in host cells and could be a promising way to control inflammation.

Influence of Epinephrine and Norepinephrine on the Risk of Metabolic Syndrome Occurrence in Workers Exposed to Hazardous Chemicals (유해화학물질 노출 근로자의 대사증후군 발생위험에 대한 epinephrine과 norepinephrine의 영향)

  • Heo, Kyung-Hwa;Kim, Ki-Woong
    • Korean Journal of Occupational Health Nursing
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    • 제19권1호
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    • pp.88-96
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    • 2010
  • Purpose: The aim of the present study was to clarify effects of long term, low-level monocyclic aromatic hydrocarbons exposure (MAHs) such as styrene, toluene and xylene on physiological levels of epinephrine (EP) and norepinephrine (NEP) and these hormones influences diagnosis indices of metabolic syndrome (MetS). Methods: Blood pressure and serum biochemical parameters were measured using digital sphygmomanometer and autochemical analyzer. EP and NEP were analyzed by using ELISA kit and exposure level of MAHs was measured by NIOSH method. Results: The differences of general characteristics such as age, smoking and drinking habits in both groups were not significantly different except working hours per day. In exposed workers, exposure levels of MAHs showed very low concentrations. Serum HDL-cholesterol concentration was significantly higher in exposed group, but concentration of NEP was significantly higher in control group. On multiple logistic regression analysis for the diagnosis indices of MetS, EP was WC (OR=0.970), NEP was blood pressure (OR=1.002) and MAHs exposure were significantly associated with HDL-cholesterol (OR=0.257), fasting glucose (OR=3.028) and MetS (OR=0.372). Conclusion: These findings suggest that the chronic exposure of low level MAHs maycontribute to glucose metabolism and induction of MetS. And also, changes of EP and NEP levels by exposure of MAHs affect blood pressure.