• Title/Summary/Keyword: enzyme-linked immunosorbent assay(ELISA)

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Effect of Tetracycline Analogues on The Activity of Matrix Metalloproteinase-3 in Gingival Fibroblasts (테트라사이클린계 약물이 치은섬유아세포내 MMP-3의 활성도에 미치는 영향)

  • Cho, Jong-Hee;Kim, Sang-Mok;Kim, Byung-Ock;Han, Kyung-Yoon
    • Journal of Periodontal and Implant Science
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    • v.29 no.3
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    • pp.677-693
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    • 1999
  • Extracellular matrix component is degraded by enzymes of thematrix metalloproteinases(MMPs). MMPs are produced by both hemopoietic and structural cells. Increased activity of MMP-3 in periodontium is strongly associated with inflammatory periodontal disease. The purpose of the present study was to determine the effect of tetracycline analogues on the activity of MMP-3. Tetracycline-HCl, doxycycline-HCl, and minocycline-HCl were applied to huamn gingival fibroblasts at various concentrations of 10, 25, 50, 100, 200${\mu}g$/ml, and 1 hour later IL-$1{\beta}$ of 25ng/ml was added. After incubation for 24 hours the cells were reacted by enzyme-linked immunosorbent assay using proMMP-3 ELISA kit. The optical density was measured by microwell plate reader at 450nm. The relative activity of MMP-3 was calculated as the percentage of the optical density of each experimental group to that of the control. The difference of the optical density and the relative activity of MMP-3 between the experimental groups and the control wasstatistically analyzed by one way ANOVA. The results were as follows: 1. Tetracycline-HCl showed the tendency to inhibit the activity of MMP-3 at the concentration lower than 25${\mu}g$/ml, but increased significantly the activity of MMP-3 at the concentration of 200${\mu}g$/ml(p<0.05). 2. Doxycycline-HCl inhibited significantly the activity of MMP-3 at the concentration lower than 100${\mu}g$/ml, but increased significantly the activity of MMP-3 at the concentration of 200${\mu}g$/ml(p<0.05). 3. Minocycline-HCl inhibited the activity of MMP-3 at the concentration in the range of 10 to 200${\mu}g$/ml. Within the limit of the present study, the above results suggested that the low concentration of tetracycline analogues could inhibit the activity of MMP-3 induced by IL-$1{\beta}$ in human gingival fibroblasts.

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Transforming Growth Factor-$\beta$ is a Possible Paracrine Mediator in the Human Endometrial Decidualization (인간자궁내막의 탈락막화 (Decudualization)에 있어서 TGF-$\beta$ (Transforming Growth Factor-$\beta$)의 역할)

  • Park, Dong-Wook;Choi, Dong-Soon;Kim, Mi-Ran;Hwang, Kyung-Joo;Jo, Mi-Yeong;Ahn, Seong-Hee;Min, Churl-K.;Ryu, Hee-Sug
    • Clinical and Experimental Reproductive Medicine
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    • v.30 no.1
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    • pp.65-75
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    • 2003
  • Objectives: To investigate the role of TGF (Transforming growth factor-$\beta$) involved in the paracrinic communication during decidualization between UEC (uterine epithelial cells) and USC (uterine stromal cells), we have employed a co-culture system composed of human endometrial epithelial and stromal cells in defined hormonal conditions. Design: In the co-culture, endometrial epithelial cells cultured in the matrigel-coated cell culture insert are seeded on top of the endometrial stromal cells cultured within a collagen gel. The co-culture was maintained for 48 hours under the following hormonal conditions: progesterone dominant condition (100 nM P4 and 1 nM E2) or estrogen-dominant condition (100 nM E2 and 1 nM P4). 10 ng/ ml HGF and/or 10 ng/ml TGF-$\beta$1 are added. Methods: RT-PCR is utilized to detect mRNAs quantitatively. Enzyme-linked immunosorbent assay (ELISA) and immunohistochemical staining are utilized to detect proteins in the tissue. Results: Prolactin mRNA is expressed in the co-cultured stromal cells under the progesterone dominant condition. TGF-$\beta$1 and its receptors are expressed in both the co-cultured epithelial and stromal cells irrespective of the steroid present, which is in contrast with no or negligible expression of TGF-$\beta$1 or its receptor in cells separately cultured. Both estrogen and progesterone significantly elevate the concentration of hepatocyte growth factor (HGF) in the conditioned medium of the co-culture with the value of 4, 325 pg/ml in E2-dominant and 2, 000 pg/ml in P4-dominant condition compare to 150 pg/ml in no hormone. In separately cultured stromal cells, administration of HGF induces the expression of TGF receptor 1 in both hormonal conditions, but induction of TGF receptor 2 is only manifest in the P4-dominant condition. Administration of TGF-$\beta$ and HGF directly induce the decidualization marker prolactin mRNA in separately cultured stromal cells. Conclusion: It is likely that steroid hormones induces prolactin mRNA indirectly by promoting the cell to cell communication between the stromal and the epithelial cells. TGF-$\beta$ and HGF are two possible paracrine mediators in the human endometrial decidualization.

The Efficacy of Saururus chinensis on Cervical Cancer Cells : The Inhibitory Effect on the Function of E6 and E7 Oncogenes of HPV Type 16 (삼백초 추출물의 자궁경부암세포 억제 효능)

  • Chung, Yeon-Gu;Lee, Hae-Sook;Lee, Kyung-Ae;Joung, Ok;Oh, Won-Keun;Kim, Kwang-Dong;Lim, Jong-Seok;Moon, Ja-Young;Cho, Yong-Kweon;Park, Sue-Nie;Yoon, Do-Young
    • YAKHAK HOEJI
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    • v.46 no.6
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    • pp.426-432
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    • 2002
  • Cervical cancer is one of the leading causes of female death from cancer worldwide with about 500,000 deaths per year. A strong association between certain human papilloma viruses (HPV type 16 and 18) and cervical cancer has been well known. An extract of Saururus chinensis, named as PE-46, has been used to investigate whether this agent has the ability of inhibiting the oncogenes E6 and E7 of HPV type 16. PE-46 inhibited the proliferation of human cervical cancer cell lines in a dose response manner. PE-46 also inhibited the in vitro binding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53. In addition, PE-46 inhibited the in vitro binding of E7 and Rb which is essential tumor suppressor for the control of cell cycle. The levels of mRNA for E6 and E7 were also decreased by PE-46. SiHa cells treated with PE-46 induced G0/G1 arrest, resulting in inhibition of growth. Our study showed that the PE-46 can inhibit the cervical carcinomas via both inhibition of bindings between oncogenes and tumor suppressors, and inhibition of G1longrightarrowS transition. PE-46 inhibited the oncogenecity of E6 and E7 of HPV 16 type, thus could be used as a putative modulating agent for the treatment of cervical carcinomas caused by HPV.

The Effect of Pilocarpine-containing Chewing Gum on Anti-microbial Components in Whole Saliva of Xerostomic Patients (구강건조증 환자에서 필로카핀 함유 껌 사용에 따른 전타액내 항균성분의 변화)

  • Park, Moon-Soo;Lee, Sung-Woo;Chung, Sung-Chang;Kim, Young-Ku;Yum, Kwang-Won
    • Journal of Oral Medicine and Pain
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    • v.24 no.4
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    • pp.347-359
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    • 1999
  • The purpose of this study was to investigate the effect of pilocarpine containing chewing gum on anti-microbial components in whole saliva of xerostomic patients, The objective xerostomic patients were instructed to use 5mg-pilocarpine containing chewing gum for 20minutes three times per day, and the author measured the flow rates of unstimulated whole saliva and stimulated whole saliva at the beginning the treatment, 1,2,3, and 4 weeks after. The concentration and flow rate of anti-microbial components in whole saliva were quantitated by enzyme-linked immunosorbent assay(ELISA). The obtained results were as follows: 1. There were significant increase in the unstimulated and stimulated whole salivary flow rate after using pilocarpine-containing chewing gum in xerostomic patients. 2. The concentrations of IgA in the unstimulated and stimulated whole saliva showed increasing pattern but, no significant changes, arid the flow rates of IgA in the unstimulated and stimulated whole saliva showed significant increase after using pilocarpine-containing chewing gum in xerostomic patients. 3. The concentrations of IgM in the unstimulated and stimulated whole saliva showed increasing pattern but, no significant changes, and the flow rates of IgM in the unstimulated and stimulated whole saliva showed significant increase after using pilocarpine-containing chewing gum in xerostomic patients. 4. The concentrations of lactoferrin in the unstimulated and stimulated whole saliva showed no significant changes, and the flow rates of lactoferrin in the unstimulated and stimulated whole saliva showed significant increase after using pilocarpine-containing chewing gum in xerostomic patients. 5. The concentrations of lysozyme in the unstimulated and stimulated whole saliva showed no significant changes, and the flow rates of lysozyme in the unstimulated whole saliva showed significant increase, but in stimulated whole saliva showed no significant changes after using pilocarpine-containing chewing gum in xerostomic patients.

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A Case of Paragonimiasis in the Abdominal Subcutaneous Tissue with Pleural Effusion (흉막 삼출을 동반한 복부 피하 지방 조직의 폐흡충증 이소기생 1례)

  • Im, So Hi;Shin, Sung Hwan;Song, Myung Jun;Kim, Jin Woo;Kim, Seung Joon;Lee, Sook Young;Kim, Young Kyoon;Park, Sung Hak
    • Tuberculosis and Respiratory Diseases
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    • v.56 no.5
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    • pp.550-554
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    • 2004
  • A paragonimiasis infestation is caused by the paragonimus species. It is commonly found in the lung but has also been found to exist extrapulmonary infestations including cerebral, spinal, subcutaneous, hepatic, splenic, abdominal, urinary, and gynecologic infestation. On the other hand, a cutaneous infestation is extremely rare. Human infestation is caused by ingesting raw or undercooked intermediate hosts. Because paragonimus westermani larva mature to an adult worm in the lung, the possibility of identifying the adult worm of paragonimus westermani at extrapulmonary region is very rare. Case : After ingesting a fresh-water crab 1 month prior to the hospital visit, a 45-year old female patient was suffering from right pleuritic chest pain during that 1 month. The patient also complained of a palpable mass that was movable and migrating, and it was localized at the right upper quadrant of the abdomen. The eosinophil fraction of the white blood cell of peripheral blood and pleural fluid was elevated to 55.1% and 90%, respectively. Parasite eggs were not found in her sputum and stool examination. By using the enzyme-linked immunosorbent assay (ELISA), the paragonimus-specific IgG antibody titer was elevated to 0.28. During incisional biopsy, we were able to find the young adult worm of paragonimus westermani. We experienced the rare case of ectopic paragonimiasis with pleural effusion that was confirmed by identifying the adult worm of paragonimus westermani within the abdominal subcutaneous tissue. We report a case with brief literature reviews.

Decreased Concentration of Plasma Brain-Derived Neurotrophic Factor in Suicide Attempters (자살 시도자에서 혈장 Brain-Derived Neurotrophic Factor 농도 저하)

  • Won, Seong-Doo;Shim, Se-Hoon;Yang, Jong-Chul;Lee, Heon-Jeong;Lee, Bun-Hee;Han, Chang-Su;Kim, Kye-Hyun;Kim, Yong-Ku
    • Korean Journal of Biological Psychiatry
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    • v.12 no.2
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    • pp.189-195
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    • 2005
  • Objects:Some studies have suggested that brain-derived neurotrophic factor(BDNF), one of the most important neurotrophins, is involved in pathophysiology of depression and suicide. This study was performed to determine whether there is an abnormality in plasma BDNF concentration in suicidal attempters. Methods:The subjects were 71 suicidal attempters who visited emergency rooms in multi-medical centers. All subjects had been interviewed by using Structured Clinical Interview for DSM-IV(SCID), Hamilton Depression Rating Scale(HDRS), Young Mania Rating Scale(YMRS), and Positive And Negative Syndrome Scale(PANSS). The severity of the suicidal behavior was measured by Lethality of Suicide Attempt Rating Scale(LSARS) and Risk-Rescue Rating(RRR) system. Seventy-one age, sex, and diagnosis matched non-suicidal psychiatric patients who were consecutively admitted to a psychiatric ward during the same period recruited as psychiatric controls. They were drug-naive or drug-free at least more than 2 months. In addition, 80 healthy controls were randomly selected as normal controls. Plasma BDNF level was measured by the enzyme linked immunosorbent assay(ELISA) methods. Results:In overall F-test, differences of the plasma BDNF levels among the groups were statistically significant(F=20.226, p<0.001). In the multiple comparisons(Scheffe), while mean levels of plasma BDNF between normal controls and non-suicidal psychiatric patients were similar(p=0.984), the BDNF levels of suicidal attempters were lower than those of other two groups(p<0.001). LSARS and RRR did not reveal any significant correlations with BDNF levels in suicidal attempters. Conclusion:These results suggest that reduction of plasma BDNF level is related to suicidal behavior and BDNF level may be a biological marker of suicidal behavior.

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Protective Effect of the Herbal Combination HCE-2 on Alcohol Metabolism and Alcohol-induced Liver Injury in Ethanol-loaded Rats (한약 복합추출물 HCE-2가 알코올을 투여한 쥐의 알코올 대사 및 간기능에 미치는 영향)

  • Han, Dong-Oh;Park, Soo-Jin;Sur, Bong-Jun;Lee, Hye-Jung;Kim, Sung-Hoon;Hahm, Dae-Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.5
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    • pp.1081-1086
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    • 2007
  • This study was performed to investigate the medicinal effects of the herbal combination extracts-2 (HCE-2), consisting of Artemisia capillaris Thunb., Lonicera japonica Thunb., Prunella vulgaris var. lilacina, and Hovenia dulcis Thunb. on the alcohol-induced liver injury in rats. The rats were randomly divided into four groups: normal group (n =6), non-treated control group (n =6), saline-treated group (n =6) and the herbal combination extract (HCE-2)-treated group (n =6). The rats in the alcohol-loaded groups were orally administered with ethanol at a daily dose of 4 g/kg-body weight for 5 weeks. Thirty minutes before the ethanol injection, saline or herbal combination extracts was administered by using a gastrogavage. Blood and liver tissue samples were taken out from the hearts and livers of the rats, respectively, on 15th and 38th days. The activities of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured using an enzyme-linked immunosorbent assay (ELISA). We also investigated the protective effect of the herbal combination extracts by Hematoxylin-Eosin staining on histological sections of rat liver. In this study, the oral administration of the herbal combination extracts significantly reduced the serum levels of AST and ALT, which had been raised by alcohol-induced liver injury. Histological analysis and apparent observation of liver also showed the preventive effect of the herbal combination extracts in a chronic alcohol-induced rat model. Theses results revealed that the herbal combination extracts effectively prevented hepatic damage consequent to the chronic exposure to repetitive administration of ethanol and could be used as a primary resource of a health beverage or herbal medicine, alleviating the alcohol-induced hepatic injury and hangover symptoms.

RELATIVE SIGNAL INTENSITY OF RETRODISCAL TISSUE IN MRI, AND SYNOVIAL FLUID CONCENTRATION OF INTERLEUKIN-6, MMP-2 AND MMP-9 IN TEMPOROMANDIBULAR JOINT DISORDER (악관절질환에서 MRI 상 관절원판 후조직의 상대적 신호강도와 관절액의 Interleukin-6, MMP-2 및 MMP-9 농도)

  • Lee, Sang-Hwa;Choie, Mok-Kyun
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.5
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    • pp.399-408
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    • 2005
  • In the progression of the Temporomandibular Joint Disorder(TMD), not only deformation and perforation of disc occur. But also fibrotic adhesion and inflammatory changes to the retrodiscal tissue can be seen in addition to the condylar degenerative change (e.g. osteoarthritis). However, the correct diagnosis,?planning for appropriate treatment, and prediction of prognosis are limited, because there are no means to stage the progression of the disorder. In this study relative signal intensity of retrodiscal tissue in MRI and the synovial fluid concentration of matrix metalloproteinase-2 (MMP-2), MMP-9, and Interleukin-6(IL-6) in the 23 temporomandibular joints(TMJ), from 17 patients with TMD were evaluated as a possible diagnostic marker. The relative signal intensity of retrodiscal tissue was referenced to brain gray matter with same region of interest(ROI) size. The concentrations of MMP-2, MMP-9, and IL-6 were evaluated by Enzyme Linked Immunosorbent Assay (ELISA). The collected data were compared with condylar degenerative change, joint effusion and disc position observed in MRI. The relative signal intensity of the retrodiscal tissue was increased significantly when degenerative changes were present. In addition, there was significantly high signal intensity in the presence of a disc displaced without reduction. The concentration of IL-6 was significantly increased when condylar degenerative change was no observed. And there were no changes in the levels of IL-6 according to disc position and joint effusion measurement. Moreover, there were no significant relevance between the concentration of total MMP-2 and active MMP-9 in synovial fluid, relative to degenerative changes in the mandibular condyle, to joint effusion, and to disc position observed on MRI images. In conclusion, the relative signal intensity of the retrodiscal tissue can be regarded as a mean of diagnosing the procession of TMD in a non-invasive manner. But more additional studies are required for the levels of MMP-2. MMP-9, and IL-6 to determine their potentials as a diagnostic marker for TMD.

Anti-inflammatory Effects of Wiryeong-tang Mix Soft Extract (위령탕 혼합단미연조엑스의 항염증 효과)

  • Kim, Myoung-Jin;Kim, Se-Jin;Nam, Won-Hee;Leem, Hyun-Hee;Jeon, Young-Hee;Son, Su-Mi;Kim, Jung-Ok
    • Herbal Formula Science
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    • v.28 no.4
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    • pp.339-349
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    • 2020
  • Objectives : Wiryeong-tang (WRT) is a traditional herbal medicine used to treat kidney-related diseases. However, the anti-inflammatory and anti-gastritis effect of Wiryeong-tang was not well known. Therefore, we experimented to confirmed the anti-inflammatory and anti-gastritis effects of Wiryeong-tang. Methods : The RAW 264.7 cells were pre treated with Wiryeong-tang mix soft extract (WRT-mse; 50, 100 and 200 ㎍/mL) for 1 hrs, and then incubated with lipopolysaccharide (LPS; 500 ng/mL). Cell viability was measured by the MTT method, and nitric oxide (NO) was measured with griess reagent. In addition, pro-inflammatory cytokines were measured by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). For anti-gastritis effect in vivo, acute gastritis was induced using 150 mM HCl/60% ethanol used ICR mice. WRT-mse (133 mg/kg) was pre treated for 3 days and then treated with 150 mM HCl/60% ethanol 1 hrs later. Then gastritis was observed and inflammatory cytokines in the gastric tissue was measured. Results : The 8 marker components of the WRT-mse were determined by simultaneous analysis using HPLC. WRT-mse was not toxic and inhibited pro-inflammatory cytokines such as IL-1β, IL-6 and TNF-α at NO production, protein and mRNA levels. Also, it was confirmed that WRT-mse improved bleeding and edema in gastritis, and suppresses inflammatory cytokines. Conclusion : In summary, our results suggest that the treatment of the WRT-mse reduced and improved the 150 mM HCl/60% ethanol induced acute gastritis and the inflammation caused by LPS stimulation in RAW 264.7 cells. Therefore, this study may provide useful drug or clinical evidence for WRT-mse to prevent inflammation.

A case of subcutaneous paragonimiasis presented with pleural effusion (흉수를 동반한 피하조직 이소기생 폐흡충증)

  • Kim, Soo Young;Park, Sun Ju;Bae, Si Young;Cho, Young Kuk;Kim, Chan Jong;Woo, Young Jong;Choi, Young Youn;Ma, Jae Sook;Hwang, Tai Ju
    • Clinical and Experimental Pediatrics
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    • v.51 no.7
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    • pp.760-765
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    • 2008
  • Paragonimiasis is a parasitic infection that occurs following the ingestion of infectious Paragonimus metacercariae from crabs or crayfish. Pulmonary paragonimiasis is the most common clinical manifestation of this infection, but several ectopic paragonimiasis cases have also been reported. Among them, cases of subcutaneous paragonimiasis are rare, especially in children. We report a case of subcutaneous paragonimiasis of the right abdominal wall with pleural effusion with hepatic involvement and without abnormal pulmonary infiltration in a boy aged 2 years and 5 months. He had eaten soybean sauce-soaked freshwater crabs (kejang) 6 months prior to complaining of right abdominal wall distension. On evaluation, right pleural effusion without abnormal pulmonary infiltration was detected, as well as blood eosinophilia, an elevated serum IgE level, pleural fluid eosinophilia and a positive enzyme-linked immunosorbent assay that detected P. westermani antibody in the serum. Thoracentesis, praziquantel administration, and excision of subcutaneous lesions were performed. After treatment, the eosinophil count and serum IgE level were decreased, and the subcutaneous lesions did not recur. The frequency of paragonimiasis has decreased recently, but it is still prevalent in Korea. Paragonimiasis should be suspected if pleural fluid eosinophilia is associated with blood hypereosinophilia and a high level of serum IgE; however clinicians should obtain a thorough history of travel and food habits.