• Title/Summary/Keyword: Cell detection

Search Result 1,502, Processing Time 0.029 seconds

Effect of the Solvent Fractions of Zingiber officinale Roscoe against Thrombintreated Tumor Invasion in Human Oral Squamous Carcinoma YD-10B Cells (YD-10B 인간구강암세포주에서 생강 유기용매 분획물의 항산화, 트롬빈억제 및 thrombin에 의해 처리된 암 침윤 및 전이 억제 효과)

  • Kim, Eun-Jung;Kim, Jun-Ho
    • Journal of Life Science
    • /
    • v.26 no.11
    • /
    • pp.1289-1297
    • /
    • 2016
  • Oral squamous cell carcinoma (OSCC) is a common malignant tumor in the oral cavity, comprising up to 90% of oral cancer. Oral cancer is characterized by a marked tendency of local invasiveness and is good for early detection and treatment; therefore, it is recognized as a good model for cancer prevention. The present study investigated the antioxidant, thrombin inhibitory, and anti-invasive activities of the solvent fractions of Zingiber officinale Roscoe. Samples were fractionated into hexane, chloroform, ethyl acetate, butanol, and water fractions, and each of these was assayed individually. The water fraction showed the highest extraction yield at 9.79%(w/w). Anti-oxidative activity was analyzed by DPPH assay. Thrombin inhibitory activity was used to analyze thrombin inhibitor assay. Cell viability was detected by the MTS assay. The activity and mRNA expression of MMP-2 and MMP-9 in human oral squamous carcinoma YD-10B cells were examined by zymography and RT-PCR. The antioxidative activities of hexane and water fractions were 92.38% and 92.96%, respectively. In the thrombin inhibitory activity test, water fraction was the highest, with a value of 65.86%. MMP-2/-9 activation was increased in phorbol 12-myristate 13-acetate (PMA)-induced YD-10B cells. MMP-9 activation was increased in thrombin-treated YD-10B cells. In PMA- or thrombin-treated YD-10B cells, the increased mRNA expression and protein activation of MMP-2/-9 were significantly inhibited in the hexane fraction. Therefore, the hexane fraction obtained from a Zingiber officinale Roscoe water extract is a promising therapeutic anti-invasive agent in oral cancer.

STIMULATION OF OSTEOBLASTIC PHENOTYPES BY STRONTIUM IN PERIOSTEAL-DERIVED CELLS (골막기원세포에서 strontium에 의한 조골세포 표현형의 활성)

  • Kim, Shin-Won;Kim, Uk-Kyu;Park, Bong-Wook;Hah, Young-Sool;Cho, Hee-Young;Kim, Jung-Hwan;Kim, Deok-Ryong;Kim, Jong-Ryoul;Joo, Hyun-Ho;Byun, June-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
    • /
    • v.32 no.3
    • /
    • pp.199-206
    • /
    • 2010
  • This study investigated the effects of strontium on osteoblastic phenotypes of cultured human periostealderived cells. Periosteal tissues were harvested from mandible during surgical extraction of lower impacted third molar. Periosteal-derived cells were introduced into cell culture. After passage 3, the periostealderived cells were further cultured for 28 days in an osteogenic induction DMEM medium supplemented with fetal bovine serum, ascorbic acid 2-phosphate, dexamethasone and at a density of $3{\times}10^4$ cells/well in a 6-well plate. In this culture medium, strontium at different concentrations (1, 5, 10, and 100 ${\mu}g$/mL) was added. The medium was changed every 3 days during the incubation period. We examined the cellular proliferation, histochemical detection and biochemical measurements of alkaline phosphatase (ALP), the RT-PCR analysis for ALP and osteocalcin, and von Kossa staining and calcium contents in the periostealderived cells. Cell proliferation was not associated with the addition of strontium in periosteal-derived cells. The ALP activity in the periosteal-derived cells was higher in 5, 10, and 100 ${\mu}g$/ml strontium-treated cells than in untreated cells at day 14 of culture. Among the strontium-treated cells, the ALP activity was appreciably higher in 100 ${\mu}g$/ml strontium-treated cells than in 5 and 10 ${\mu}g$/ml strontium-treated cells. The levels of ALP and osteocalcin mRNA in the periosteal-derived cells was also higher in strontium-treated cells than in untreated cells at day 14 of culture. Their levels were increased in a dose-dependent manner. Von Kossa-positive mineralization nodules were strongly observed in the 1 ${\mu}g$/ml strontium-treated cells at day 21 and 28 of culture. The calcium content in the periosteal-derived cells was also higher in 1 ${\mu}g$/ml strontium-treated cells at day 28 of culture. These results suggest that low concentration of strontium stimulates the osteoblastic phenotypes of more differentiated periosteal-derived cells, whereas high concentration of strontium stimulates the osteoblastic phenotypes of less differentiated periosteal-derived cells. The effects of strontium on osteoblastic phenotypes of periosteal-derived cells appear to be associated with differentiation-extent.

Cellular Analysis and Measurement of Mucin in Sputum of Chronic Airway Disease (만성기도질환의 객담세포분석과 mucin의 측정)

  • Kim, Ki-Up;Kim, Yang-Ki;Shin, Chan-Young;Kim, Do-Jin;Uh, Soo-Taek;Kim, Yong-Hoon;Ko, Kwang-Ho;Park, Choon-Sik
    • Tuberculosis and Respiratory Diseases
    • /
    • v.49 no.1
    • /
    • pp.82-92
    • /
    • 2000
  • Background : In chronic airway disease, mucus secretion is increased, but extraction of mucin, which is the main component of mucus secretion, is a very complicated and limited in clinical use. Recently, monoclonal antibody for mucin was developed for possible clinical use. In this study, cellular analysis and quantification of respiratory mucin in sputum of patients with chronic airway diseases were performed. Method : Sputum was collected from patients with asthma(n=33), bronchiectasis(n=8) or chronic bronchitis (n=13) by spontaneous expectoration or by hypertonic saline induction. Collected sputums was treated by 0.1% dithiotreitol to dissociate the disulfide bond of the mucus and filtered through a nylon gauze. Total cell count, viability and differential count were measured. For detection of mucin, collected samples were treated with sodium dodecyl sulfate polyacrylamide gel electrophoresis and then with monoclonal antibody(HMO2), as the primary antibody, and PAS stain. The amount of mucin was measured with ELISA by HMO2. Correlation with clinical information, cellular analysis, and amount of measured mucin were analyzed. Results : Total cell counts of sputum were significantly increased in patients with bronchiectasis but viability remained the same. Eosinophils were significantly increased in patients with asthma, neutrophils in bronchiectasis chronic bronchitis, respectively (p<0.05). The results of Western blotting and PAS staining confirmed the presence of glycoproteins and matched? with mucin. The amounts of mucin measured by ELISA were not significantly different among the disease groups. Significant correlation was identified between the amount of mucin and viability(r=-0.482, p<0.05). Conclusion : Inflammatory cells in the sputum of those with chronic airway disease were different for each disease type. Measurement of mucin by ELISA via monoclonal antibodies may be a simple method for the evaluation of chronic airway disease.

  • PDF

Efficacy of Positron Emission Tomography in Diagnosing Pulmonary Tumor and Staging of Lung Cancer : Comparing to Computed Tomography (폐종양과 폐암의 병기결정에 대한 양전자단층촬영(PET)의 유용성 -전산화단층촬영 (CT)과의 비교-)

  • 김오곤;조중행;성숙환
    • Journal of Chest Surgery
    • /
    • v.36 no.2
    • /
    • pp.79-85
    • /
    • 2003
  • Diagnosing and determining the stage of lung cancer by means of positron emission tomography (PET) ha.. been proven valuable because of the limitations of diagnosis by computed tomography (CT). We compared the efficacy of PET with that of CT in diagnosing pulmonary tumor and staging of lung cancer Material and Method: We performed F-18 FDG PET to determine the malignancy and the staging on patients who have been suspicious or were diagnosed as lung cancer by chest X-ray and CT. The findings of PET and of CT of 41 patients (male, 29: female, 12: mean age, 59) were compared with pathologic findings obtained from a mediastinoscopy and thoracotomy. Result: Out of 41 patients, 35 patients had malignant lesions (squamous cell carcinonla 19 cases, adenocarcinoma 14 cases, adenosquamous cell carcinoma 2 cases) and 6 patients had benign lesions. Diagnosing of lung cancer, the sensitivity, specificity and accuracy of CT and PET were the same for two method and the numbers were 100%, 50%, and 92.7% respectively. Eighteen LN groups out of 108 mediastinal LN groups who recieved histologic examination proved to be malignant. Pathologic lymph node (LN) stage was N0-Nl 31 cases, N2 8 cases, N3 2 cases. The correct identification of the nodal staging with CT, PET scans were 31 cases (75.6%), 28 cases (68.3%) respectively. The LN group was underestimated in each 6 cases of CT and PET. In 4 cases of CT and 7 cases of PET, they were overestimated in compare to histologic diagnosis. In the detection of mediastinal LN groups invasion, the sensitivity, specificity and accuracy of CT were 39.8 %, 93.3 %, and 84.3 % respectively. For PET, they were 61.1 %, 90.0 %, and 85.2 %. When two methods considered together (CT+PET), they were increased to 77.8 %, 93.3 %, and 90.7 % respectively. Conclusion: PET appears to be similar to CT in the diagnosis and the nodal taging of pulmonary tumor. Two tests may stage patients with lung cancer more accurately than CT alone.

Chromosomal Localization and Mutation Detection of the Porcine APM1 Gene Encoding Adiponectin (Adiponectin을 암호화하는 돼지 APM1 유전자의 염색체상 위치파악과 돌연변이 탐색)

  • Park, E.W.;Kim, J.H.;Seo, B.Y.;Jung, K.C.;Yu, S.L.;Cho, I.C.;Lee, J.G.;Oh, S.J.;Jeon, J.T.;Lee, J.H.
    • Journal of Animal Science and Technology
    • /
    • v.46 no.4
    • /
    • pp.537-546
    • /
    • 2004
  • Adiponectin is adipocyte complement-related protein which is highly specialized to play important roles in metabolic and honnonal processes. This protein, called GBP-28, AdipoQ, and Acrp30, is encoded by the adipose most abundant gene transcript 1 (APM1) which locates on human chromosome 3q27 and mouse chromosome 16. In order to determine chromosomal localization of the porcine APM1, we carried out PCR analysis using somatic cell hybrid panel as well as porcine whole genome radiation hybrid (RH) panel. The result showed that the porcine APM1 located on chromosome 13q41 or 13q46-49. These locations were further investigated with the two point analysis of RH panel, revealed the most significant linked marker (LOD score 20.29) being SIAT1 (8 cRs away), where the fat-related QTL located. From the SSCP analysis of APM1 using 8 pig breeds, two distinct SSCP types were detected from K~ native and Korean wild pigs. The determined sequences in Korean native and Korean wild pigs showed that two nucleotide positions (T672C and C705G) were substituted. The primary sequence of the porcine APM1 has 79 to 87% identity with those of human, mouse, and bovine APM1. The domain structures of the porcine APM1 such as signal sequence, hypervariable region, collagenous region. and globular domain are also similar to those of mammalian genes.

Dynamin II Expression and Morphological Comparison of NIH3T3 and NIH3T3 (ras) Cells (NIH3T3와 NIH3T3(ras) 세포에서 Dynamin II 발현 및 형태적 비교)

  • Lee, Chul-Woo;Kim, Su-Gwan;Choi, Jeong-Yun;Choi, Baik-Dong;Bae, Chun-Sik;Jeong, Soon-Jeong;Jeong, Moon-Jin
    • Applied Microscopy
    • /
    • v.35 no.3
    • /
    • pp.121-128
    • /
    • 2005
  • It has been known that ras signaling transduction leads to cell proliferation and migration including various adaptor molecules. Dynamin protein has been implicated in the formation of nascent vesicles in both the endocytic and secretory pathways. Dynamin was classified into three isoforms: dynamin I is only expressed in neuronal tissue, dynamin II is expressed ubiquitously in all tissue but that of dynamin III is confined to testis. We have reported in previous study that Grb2, binding to ras, was associated with dynamin II in NIH3T3 cells. Therefore we have tried to identify the relative expression of dynamin II according to overexpressed ras protein in ras oncogene transfected cells (NIH3T3 (ras)). For the detection of differential expression of dynamin II, we have used immunofluorescent staining and western blot methods in NIH3T3 and NIH3T3 (ras) cells. Next we have described the morphological differences between NIH3T3 and NIH3T3 (ras) cells using SEM and TEM. From these experiments dynamin II was highly expressed in NIH3T3 (ras) cells. NIH3T3 cells was transformed to more spindle shape with many cell process by transfection of ras oncogene. Moreover dynamin II was more concentrated in endocytotic membrane of the NIH3T3 (ras) cells compared to that of NIH3T3 cells. The present results suggested that dynamin II may involve the intermediate messenger in Ras signaling transduction pathway.

Increased Protein of the Secretory Leukocyte Pretense Inhibitor (SLPI) and the Expression of Growth Factors in NIH3T3 Cells by LPS Stimulation (NIH3T3 세포주에서 LPS자극에 의한 분비백혈구단백분해효소억제제 (SLPI)의 단백질증가와 성장인자들의 발현)

  • Lee, Sang-Hwa;Choi, Baik-Dong;Jeong, Soon-Jeong;Jang, Hyun-Seon;Kim, Byung-Ock;Lim, Do-Seon;Park, Joo-Cheol;Wang, Guan-Lin;Jeong, Moon-Jin
    • Applied Microscopy
    • /
    • v.36 no.3
    • /
    • pp.165-172
    • /
    • 2006
  • Secretory leukocyte protease inhibitor (SLPI) involves tissue protection against the destructive action of neutrophil elastase at the site of inflammation. Several studies on new functions of SLPI have demonstrated that SLPI may play a primary role in innate immunity than protease inhibitor, To identify the function of SLPI by lipopolysaccharide (LPS) stimulation in the embryonic fibroblast (NIH3T3) cells. we studied the expression of SLPI compared to other growth factors involving the LPS treatment. To address this, we performed the reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of the SLPI and some growth factors such as VEGF. bFGF, and PDGF-BB after LPS stimulation. NIH3T3 cells were exposed 100 ng/mL Escherichia coli LPS for 30min, 60min, 90min, 24h, and 48h, respectively. The result of RT-PCR showed that SLPI and VEGF mRNA was expressed strongly in NIH3T3 without related to LPS stimulation. mRNA of bFGF was weakly expressed such as the expression of the control. PDGF mRNA expression gradually increased follows at time course. However, SLPI protein level was increased in lysates and culture medium by LPS stimulation. Phase contrast microscopic and scanning electron microscopic observation showed that the increased cell number and cytoplasmic enlargement of the NIH3T3 cells. Therefore, it suggests that the LPS upregulates SLPI expression in NIH3T3 cells. Moreover, secreted SLPI may stimulate cell proliferation and migration.

Anti-oxidative and Anti-inflammatory Activities of Fermented Turmeric (Curcuma longa L.) by Rhizopus oryzae (Rhizopus oryzae으로 발효한 울금의 항산화 및 항염효과)

  • Kim, Eun-Ju;Song, Bit-Na;Jeong, Da-Som;Kim, So-Young;Cho, Yong-Sik;Park, Shin-Young
    • Journal of Life Science
    • /
    • v.27 no.11
    • /
    • pp.1315-1323
    • /
    • 2017
  • Turmeric is a rhizomatous herbaceous perennial plant (Curcuma longa (CL)) of the ginger family, Zingiberaceae. A yellow-pigmented fraction isolated from the rhizomes of CL contains curcuminoids belonging to the dicinnamoyl methane group. Curcumin is an important active ingredient responsible for the biological activity of CL. However, CL is not usually used as a food source due to its bitter taste. The present study was designed to determine the effect of the CL fermented by Rhizopus oryzae (FCL) on pro-inflammatory factors such as nuclear factor ${\kappa}B$ ($NF-{\kappa}B$), tumor necrosis factor alpha ($TNF-{\alpha}$), interleukin-6 (IL-6), nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-induced RAW 264.7 cell line. The cell viability was determined by MTT assay. To evaluate the anti-inflammatory effect of FCL 80% EtOH extracts, IL-6 and $TNF-{\alpha}$ were measured by ELISA kit. Also, the amount of $NO/PGE_2/NF-{\kappa}B$ was measured using the $NO/PGE_2/NF-{\kappa}B$ detection kit and the iNOS/COX-2 expression was measured by Western blotting. The results showed that the FCL reduced NO, $PGE_2$, iNOS, COX-2, $NF-{\kappa}B$, IL-6 and $TNF-{\alpha}$ production without cytotoxicity. These results suggest that FCL extracts may be a developed the functional food related to anti-inflammation due to the significant effects on inflammatory factors.

Primary Synchronous Lung Cancer Detected using Autofluorescence Bronchoscopy (자가 형광 기관지 내시경을 이용한 동시성 원발성 폐암의 진단)

  • Kwon, Sun Jung;Lee, Yun Seun;Joung, Mi Kyong;Lee, Yu Jin;Jang, Pil Soon;Lee, Jeung Eyun;Chung, Chae Uk;Park, Hee Sun;Jung, Sung Soo;Kim, Sun Young;Kim, Ju Ock
    • Tuberculosis and Respiratory Diseases
    • /
    • v.60 no.6
    • /
    • pp.645-652
    • /
    • 2006
  • Objective: Patients with lung cancer have a relative high risk of developing secondary primary lung cancers. This study examined the additional value of autofluorescence bronchoscopy (AFB) for diagnosing synchronous lung cancers and premalignant lesions. Methods: Patients diagnosed with lung cancer from January 2005 to December 2005 were enrolled in this study. The patients underwent a lung cancer evaluation, which included white light bronchoscopy (WLB), followed by AFB. In addition to the primary lesions, any abnormal or suspicious lesions detected during WLB and AFB were biopsied. Results: Seventy-six patients had non-small cell lung cancer (NSCLC) and 23 had small cell lung cancer (SCLC). In addition to the primary lesions, 84 endobronchial biopsies were performed in 46 patients. Five definite synchronous cancerous lesions were detected in three patients with initial unresectable NSCLC and in one with SCLC. The secondary malignant lesions found in two patients were considered metastatic because of the presence of mediastinal nodes or systemic involvement. One patient with an unresectable NSCLC, two with a resectable NSCLC, and one with SCLC had severe dysplasia. The detection rate for cancerous lesions by the clinician was 6.0% (6/99) including AFB compared with 3.0% (3/99) with WLB alone. The prevalence of definite synchronized cancer was 4.0% (4/99) after using AFB compared with 2.0% (2/99) before, and the staging-up effect was 1.0% (1/99) after AFB. Since the majority of patients were diagnosed with advanced disease, the subjects with newly detected cancerous lesions did not have their treatment plans altered, except for one patient with a stage-up IV NSCLC who did not undergo radiotherapy. Conclusions: Additional AFB is effective in detecting early secondary cancerous lesions and is a more precise tool in the staging workup of patients with primary lung cancer than with WLB alone.

The Effectiveness of Visual Field C10-2 in the Early Detection of Glaucoma with Parafoveal Scotoma (중심부 시야결손을 보이는 초기 녹내장 환자의 진단에서 자동시야계 C10-2의 유용성)

  • Hwang, Bo Een;Park, Hae-Young Lopilly;Park, Chan Kee
    • Journal of The Korean Ophthalmological Society
    • /
    • v.58 no.3
    • /
    • pp.321-326
    • /
    • 2017
  • Purpose To identify the correspondence between the central sensitivity of several visual field (VF) tests and ganglion cell inner plexiform layer (GC-IPL) thickness in early glaucoma patients with parafoveal scotoma. Methods Fifty-seven eyes from 57 patients with glaucomatous optic neuropathy and parafoveal scotoma were analyzed using the standard automated perimetry (SAP) C10-2 test, the SAP C24-2 test, and the frequency doubling technology perimetry (FDT) C24-2 test. The correlation between the VF central sensitivity and the GC-IPL thickness from macular scans via optical coherence tomography was analyzed. Results The central sensitivity was $27.51{\pm}5.43dB$, $27.39{\pm}5.05dB$, and $22.09{\pm}5.08dB$ for SAP C24-2, SAP C10-2, and FDT C24-2, respectively. Mean GC-IPL thickness was $70.2{\pm}8.5{\mu}m$. Using regression analysis, the value of log $R^2$ between the logarithmic central sensitivity and GC-IPL thickness was 0.498, and the linear $R^2$ between the antilogarithmic central sensitivity and GC-IPL thickness in SAP C10-2 was 0.486, and both were statistically significant (p < 0.05). This relationship was stronger in early glaucoma patients compared to late glaucoma patients using SAP C10-2. Conclusions The structure-function relationship between GC-IPL thickness and central sensitivity was better with SAP C10-2, especially in early glaucoma patients, compared to other VF modalities.