• Title/Summary/Keyword: Normal tissue cells

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Development of a Primary Tissue Culture Method having Greater Reliability than Isolated Cell Cultures - Steroid-Responsiveness of Uterine Myometrial and Myomatous(Leiomyomatous) Cells (자궁근종세포의 최적 초기배양 조건 확립 - 정상 자궁근세포와 자궁근종세포의 스테로이드에 대한 반응)

  • Lee, Eun-Ju;Bajracharya, Prati;Hyun, Jin-Hee;Kim, Hang-Jin;Song, Gun-Ho;Cho, Kyung-Hyun;Lee, Dong-Mok;Lee, Taek-Hoo;Chun, Sang-Sik;Choi, In-Ho
    • Development and Reproduction
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    • v.11 no.3
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    • pp.205-217
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    • 2007
  • As an initial step toward better understanding of the molecular mechanism of estrogen-dependent growth in myoma, an optimal primary cell culture condition has been developed and examined by this study. Myoma and myometrium were cultured by two different methods. Culture stability and $E_2$-responsiveness in stable culture were studied. The culture of digested tissue pieces(Method 2) was found to be a stable culture method for the myoma and myometrium showing a favorable response to estrogen. mRNA expression of PR, IGF-1 and IGF-1 receptor genes was enhanced by $E_2$. The gene responses to $E_2$ were higher in myoma compared with myometrium. Moreover, these responses were more expressive in tissues than in the surrounding cells in primary culture of normal myometrium and myoma, implying a vital role of cell communication through the extracellular matrix in maintaining the estrogen-responsiveness. The development of an improved cell culture system for myoma provides an in vitro tool to further investigate the basis of the tumor formation.

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Influence of covering treatment on the incidence of frost injury in chinese cabbage during winter season (피복처리가 월동배추 동해 발생에 미치는 영향)

  • Lee, Sang Gyu;Choi, Chang Sun;Lee, Hee Ju;Jang, Yoon Ah;Do, Kyung Ran
    • Korean Journal of Agricultural Science
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    • v.41 no.3
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    • pp.163-167
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    • 2014
  • The average temperatures for year and winter season have been risen by $0.7{\circ}C$ and $1.4{\circ}C$, respectively, during the last 30 years. Recently abnormal climate phenomena occurred frequently results in severe loss of vegetable crops grown in Korea. Specially, Chinese cabbages grown in the southern area of Korea are often significantly affected by sudden cold waves during winter season before harvest. This experiment was conducted to find out a potential role of covering materials on the protection of frost damage of 'Bularm' chinese cabbage in the winter season. The lowest temperature was $-15.8^{\circ}C$ in non-covering, $-8.1^{\circ}C$ in the PE film covering and $-4.6^{\circ}C$ in the non-woven fabric covering with PE film, respectively. The cumulative times below $4.0^{\circ}C$ were 145.5 hours for the non-covering treatment, 94 hours in the PE film covering and 14.5 hours in the non-woven fabric covering with PE film, respectively. The symptoms of frost damage were severe at non-covering chinese cabbages compared to polyethylene film (PE) non-woven fabric with PE covering ones. Microscopic studies showed the normal anatomical structure of palisade and spongy tissue of cabbage leaves covered with non-woven fabric with PE film. Leaf cells, however, were slightly damaged in cabbages covered with PE film alone, and both palisade and spongy cells were were completely collapsed in uncovered cabbages. The result of this study suggests that chinese cabbages is required to be covered with non-woven fabric with PE film to minimize the frost damage by sudden cold wave below $-7^{\circ}C$.

xCyp26c Induced by Inhibition of BMP Signaling Is Involved in Anterior-Posterior Neural Patterning of Xenopus laevis

  • Yu, Saet-Byeol;Umair, Zobia;Kumar, Shiv;Lee, Unjoo;Lee, Seung-Hwan;Kim, Jong-Il;Kim, SungChan;Park, Jae-Bong;Lee, Jae-Yong;Kim, Jaebong
    • Molecules and Cells
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    • v.39 no.4
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    • pp.352-357
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    • 2016
  • Vertebrate neurogenesis requires inhibition of endogenous bone morphogenetic protein (BMP) signals in the ectoderm. Blocking of BMPs in animal cap explants causes the formation of anterior neural tissues as a default fate. To identify genes involved in the anterior neural specification, we analyzed gene expression profiles using a Xenopus Affymetrix Gene Chip after BMP-4 inhibition in animal cap explants. We found that the xCyp26c gene, encoding a retinoic acid (RA) degradation enzyme, was upregulated following inhibition of BMP signaling in early neuroectodermal cells. Whole-mount in situ hybridization analysis showed that xCyp26c expression started in the anterior region during the early neurula stage. Overexpression of xCyp26c weakly induced neural genes in animal cap explants. xCyp26c abolished the expression of all trans-/cis-RA-induced posterior genes, but not basic FGF-induced posterior genes. Depletion of xCyp26c by morpholino-oligonucleotides suppressed the normal formation of the axis and head, indicating that xCyp26c plays a critical role in the specification of anterior neural tissue in whole embryos. In animal cap explants, however, xCyp26c morpholinos did not alter anterior-to-posterior neural tissue formation. Together, these results suggest that xCyp26c plays a specific role in anterior-posterior (A-P) neural patterning of Xenopus embryos.

Respiratory Protective Effect of a RML on PM10D-induced Lung Injury Mouse Model (미세먼지 유발 폐기능 손상 동물모델에서 RML의 호흡기 보호 효과)

  • Kim, Soo Hyun;Kim, Min Ju;Shin, Mi-Rae;Roh, Seong-Soo;Kim, Seung Hyung;Park, Hae-Jin
    • The Korea Journal of Herbology
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    • v.37 no.3
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    • pp.29-39
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    • 2022
  • Objective : This study is aimed to evaluate the protective effects of Rehmanniae Radix, Mori Folium, and Liriopie Tuber mixture (RML) on lung injury of Particulate matter less than 10 um in diameter and diesel exhaust particles (PM10D) mice model. Methods : To investigate the anti-inflammatory activity of RML, PM10D was diluted in aluminum hydroxide (Alum) in 7-week-old male mice and induced by Intra-Nazal-Tracheal (INT) injection method. Animal experiments were divided into 5 groups. Nor (normal mice), CTL (PM10D-induced mice with the administration of distilled water), DEXA (PM10D-induced mice with the administration of 3 mg/kg Dexamethasone), RML 100 (PM10D-induced mice treated with RML 100 mg/kg weight), and RML 200 (PM10D-induced mice treated with RML 200 mg/kg body weight). After 11 days administration, mice were sacrificed and inflammation-related immune cells in broncho-alveolar lavage fluid (BALF) were analyzed. Inflammation-related biomarkers were also analyzed in blood and lungs. Lung tissue was observed through histological examination. Results : In the PM10D induced model, the PML showed decreases in CXCL-1 and IL-17A in BALF. Expression of inflammatory cytokines and cough-related mRNA genes was significantly decreased in serum and lung tissue. The mixture treatment of RML significantly improved the immune related cells in the serum. In addition, histological observations showed a tendency to decrease the severity of lung injury. Conclusions : Overall, these results confirmed the respiratory protective effect of the RML mixture in a model of lung injury induced by air pollution (PM10+DEP), suggesting that it is a potential treatment for respiratory damage.

Decay-Accelerating Factor Differentially Associates With Complement-Mediated Damage in Synovium After Meniscus Tear as Compared to Anterior Cruciate Ligament Injury

  • V. Michael Holers;Rachel M. Frank;Michael Zuscik;Carson Keeter;Robert I. Scheinman;Christopher Striebich;Dmitri Simberg;Michael R. Clay;Larry W. Moreland;Nirmal K. Banda
    • IMMUNE NETWORK
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    • v.24 no.2
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    • pp.17.1-17.16
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    • 2024
  • We have reported that anterior cruciate ligament (ACL) injury leads to the differential dysregulation of the complement system in the synovium as compared to meniscus tear (MT) and proposed this as a mechanism for a greater post-injury prevalence of post traumatic osteoarthritis (PTOA). To explore additional roles of complement proteins and regulators, we determined the presence of decay-accelerating factor (DAF), C5b, and membrane attack complexes (MACs, C5b-9) in discarded surgical synovial tissue (DSST) collected during arthroscopic ACL reconstructive surgery, MT-related meniscectomy, osteoarthritis (OA)-related knee replacement surgery and normal controls. Multiplexed immunohistochemistry was used to detect and quantify complement proteins. To explore the involvement of body mass index (BMI), after these 2 injuries, we examined correlations among DAF, C5b, MAC and BMI. Using these approaches, we found that synovial cells after ACL injury expressed a significantly lower level of DAF as compared to MT (p<0.049). In contrast, C5b staining synovial cells were significantly higher after ACL injury (p<0.0009) and in OA DSST (p<0.039) compared to MT. Interestingly, there were significantly positive correlations between DAF & C5b (r=0.75, p<0.018) and DAF & C5b (r=0.64 p<0.022) after ACL injury and MT, respectively. The data support that DAF, which should normally dampen C5b deposition due to its regulatory activities on C3/C5 convertases, does not appear to exhibit that function in inflamed synovia following either ACL injury or MT. Ineffective DAF regulation may be an additional mechanism by which relatively uncontrolled complement activation damages tissue in these injury states.

THE EFFECTS OF IRRADIATION ON THE MANDIBULAR BONE OF RATS ON THE LOW CALCIUM DIET (방사선조사가 저칼슘식이 백서의 하악골에 미치는 영향에 관한 실험적 연구)

  • Hwang Eui-Hwan;Lee Sang-Rae
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.22 no.2
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    • pp.241-252
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    • 1992
  • The purpose of this study was to investigate the changes of morphology and structure of bone tissue in the irradiated mandibular bone in rats which were fed a low calcium diet. In order to carry out this experimant, 64 seven-week old Sprague-Dawley strain rats weighing about 150gms were selected and equally divided into one experimental group of 32 rats and one control group with the remainder. The experimental group and the control group were then subdivided into two groups when the rats reached ten-week old, 16 were assigned rats for each subdivided group, exposed to irradiation. The two irradiation groups received a single dose of 20 Gy in the jaws area only and irradiated with a cobalt-60 teletherapy unit. The rats in the control and experimental groups were serially terminated by fours on the 3rd, the 7th, the 14th, and the 21st day after irradiation. Following termination, both sides of the dead rats mandibular bodies were removed and fixed with 10% neutral formalin. One side of the mandibular body was radiographed with a soft X-ray apparatus. Thereafter, the obtained microradiographs were observed by a light microscope. The remaining side of the mandibular bone was further decalcified and embedded in paraffin as using the general method. The specimen sectioned and stained with hematoxylin and eosin, and Rabit Anti-Human Tumor Necrosis Factor-a, observed by a light microscope. The obtained results were as follows: 1. Microradiogram revealed that thinning of the cortex and a decrease in the trabecula of the interradicular bone and mandibular body were observed and noted from the start to finish throughout the experiment in the non-irradiated rats on the low calcium diet rather than in the non-irradiated rats on the normal diet. In microscopic observations, there were marked osteolytic changes in the center of the bone marrow. 2. Microradiogram revealed that thinning of the cortex and a decrease in the trabecula of interradicular bone and mandibular body were more marked after 7 days in the irradiated rats on the low calcium diet rather than in the non-irradiated rats on the low clacium diet. In microscopic observations, osteoblasts were decreased markedly after 7 days, and a few osteoclasts were observed. 3. Microradiogram revealed that thinning of the cortex and a decrease in trabecula of interradicular bone and the mandibular body were more marked from the strart to finish troughout the experiment in the irradiated rats on the low calcium diet rather than in the irradiated rats on the normal diet. In microscopic observations, there were no osteolytic changes noted in the irradiated rats on the normal diet. 4. In immunocytochemical findings, a few bone marrow cells including Tumor Necrosis Factor were observed in the irradiated rats on the normal diet, but was not observed in the rats on the low calcium diet.

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Research on Perfusion CT in Rabbit Brain Tumor Model (토끼 뇌종양 모델에서의 관류 CT 영상에 관한 연구)

  • Ha, Bon-Chul;Kwak, Byung-Kook;Jung, Ji-Sung;Lim, Cheong-Hwan;Jung, Hong-Ryang
    • Journal of radiological science and technology
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    • v.35 no.2
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    • pp.165-172
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    • 2012
  • We investigated the vascular characteristics of tumors and normal tissue using perfusion CT in the rabbit brain tumor model. The VX2 carcinoma concentration of $1{\times}10^7$ cells/ml(0.1ml) was implanted in the brain of nine New Zealand white rabbits (weight: 2.4kg-3.0kg, mean: 2.6kg). The perfusion CT was scanned when the tumors were grown up to 5mm. The tumor volume and perfusion value were quantitatively analyzed by using commercial workstation (advantage windows workstation, AW, version 4.2, GE, USA). The mean volume of implanted tumors was $316{\pm}181mm^3$, and the biggest and smallest volumes of tumor were 497 $mm^3$ and 195 $mm^3$, respectively. All the implanted tumors in rabbits are single-nodular tumors, and intracranial metastasis was not observed. In the perfusion CT, cerebral blood volume (CBV) were $74.40{\pm}9.63$, $16.08{\pm}0.64$, $15.24{\pm}3.23$ ml/100g in the tumor core, ipsilateral normal brain, and contralateral normal brain, respectively ($p{\leqq}0.05$). In the cerebral blood flow (CBF), there were significant differences between the tumor core and both normal brains ($p{\leqq}0.05$), but no significant differences between ipsilateral and contralateral normal brains ($962.91{\pm}75.96$ vs. $357.82{\pm}12.82$ vs. $323.19{\pm}83.24$ ml/100g/min). In the mean transit time (MTT), there were significant differences between the tumor core and both normal brains ($p{\leqq}0.05$), but no significant differences between ipsilateral and contralateral normal brains ($4.37{\pm}0.19$ vs. $3.02{\pm}0.41$ vs. $2.86{\pm}0.22$ sec). In the permeability surface (PS), there were significant differences among the tumor core, ipsilateral and contralateral normal brains ($47.23{\pm}25.45$ vs. $14.54{\pm}1.60$ vs. $6.81{\pm}4.20$ ml/100g/min)($p{\leqq}0.05$). In the time to peak (TTP) were no significant differences among the tumor core, ipsilateral and contralateral normal brains. In the positive enhancement integral (PEI), there were significant differences among the tumor core, ipsilateral and contralateral brains ($61.56{\pm}16.07$ vs. $12.58{\pm}2.61$ vs. $8.26{\pm}5.55$ ml/100g). ($p{\leqq}0.05$). In the maximum slope of increase (MSI), there were significant differences between the tumor core and both normal brain($p{\leqq}0.05$), but no significant differences between ipsilateral and contralateral normal brains ($13.18{\pm}2.81$ vs. $6.99{\pm}1.73$ vs. $6.41{\pm}1.39$ HU/sec). Additionally, in the maximum slope of decrease (MSD), there were significant differences between the tumor core and contralateral normal brain($p{\leqq}0.05$), but no significant differences between the tumor core and ipsilateral normal brain($4.02{\pm}1.37$ vs. $4.66{\pm}0.83$ vs. $6.47{\pm}1.53$ HU/sec). In conclusion, the VX2 tumors were implanted in the rabbit brain successfully, and stereotactic inoculation method make single-nodular type of tumor that was no metastasis in intracranial, suitable for comparative study between tumors and normal tissues. Therefore, perfusion CT would be a useful diagnostic tool capable of reflecting the vascularity of the tumors.

Shipyeukmiyeugi-eum Extracts Suppressed Tumor Growth through Immunomodulatory Effects on MCF-7 (십육미유기음(十六味流氣飮)의 면역활성(免疫活性)에 의한 유방암(乳房癌) 세포 성장억제효과(成長抑制效果))

  • Jung, Yeon-Chul;Park, Young-Sun;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.25 no.3
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    • pp.40-60
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    • 2012
  • Objectives: The object of this study was to observe antitumor, anticachexia and immunomodulatory effects of Shipyeukmiyeugi-eum(SYM) on human breast cancer cell, MCF-7, xenograft Balb/c nu-nu nude mice. Methods: Three different dosages of SYM-125, 250 and 500 mg/kg were orally administered once a day for 28 days from 11 days after tumor cell inoculation, and the changes on the body weights, tumor volume and weights, weights of spleen and popliteal lymph node and epididymal fat, serum IL-6 and IFN-${\gamma}$ levels, NK cell and peritoneal macrophage activities, splenic TNF-${\alpha}$, IL-$1{\beta}$ and IL-10 contents were observed. In addition, histopathological observations of apoptotic cell, spleen, popliteal lymph node and cervical brown adipose were also detected. The results were compared with a potent cytotoxic estrogen receptor antagonist, Tamoxifen 20 mg/kg treated mice. Results: Tumor volumes and weights were decreased without cytotoxic effects on the both MCF-7 and MCF-10A cells as results of all three different dosages of SYM treatment. And weights of body, spleen, popliteal lymph node, epididymal fat, serum IFN-${\gamma}$, NK cell, peritoneal macrophage activities, splenic TNF-${\alpha}$, IL-$1{\beta}$ and IL-10 contents were increased with decrease of serum IL-6. At histopathological observations, apoptotic tumor cells, spleen, popliteal lymph node and cervical brown adipose tissue were increased. That means tumor-related immunosuppress and cachexia were markedly inhibited by SYM treatment as compared with tumor-bearing mice. On the other hand, Tamoxifen showed marked cytotoxic effects against MCF-7 and MCF-10A, decreases of tumor volume and weights, and increases of apoptotic tumor cells and related decreases of tumor cell volumes, but tamoxifen markedly deteriorated the tumor-related immune-suppress and cachexia. Conclusions: The results obtained in this study suggest that SYM showed favorable anticancer effects and anticachexic effects on the MCF-7 cell xenograft through immunomodulatory effects. SYM did not induce any cytotoxic effects against both normal and cancer cells.

The Effect of Remifentanil Preconditioning on Injured Keratinocyte

  • Hong, Hun Pyo;Kim, Cheul Hong;Yoon, Ji Young;Kim, Yong Deok;Park, Bong Soo;Kim, Yong Ho;Yoo, Ji Uk
    • Journal of The Korean Dental Society of Anesthesiology
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    • v.14 no.3
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    • pp.157-165
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    • 2014
  • Background: Incisional site of surgical operation become transient ischemic state and then occur reoxygenation due to vasodilatation by inflammatory reaction, the productive reactive oxygen species (ROS) give rise to many physiologic results. Apoptosis have major role on elimination of inflammatory cell and formation of granulation tissue in normal wound healing process. Remifentanil can prevent the inflammatory response and can suppress inducible nitric oxide synthase expression in a septic mouse model. After cardiopulmonary bypass for coronary artery surgery, remifentanil can also inhibit the release of biomarkers of myocardial damage. Here we investigated whether remifentanil pretreatment has cellular protective effect against hypoxia-reoxygenation in HaCaT human keratinocytes, if so, the role of apoptosis and autophagy on this phenomenon. Methods: The HaCaT human keratinocytes were exposed to various concentrations of remifentanil (0.01, 0.05, 0.1, 0.5 and 1 ng/ml) for 2 h before hypoxia (RPC/HR group). These cells were cultured under 1% oxygen tension for 24h at $37^{\circ}C$. After hypoxia, to simulate reoxygenation and recovery, the cells were reoxygenated for 12 h at $37^{\circ}C$. 3-MA/RPC/HR group was treated 3-methyladenine (3-MA), autophagy inhibitor for 1h before remifentanil treatment. Cell viability was measured using a quantitative colorimetric assay with thiazolyl blue tetrazoliumbromide (MTT, amresco), showing the mitochondrial activity of living cells. To investigate whether the occurrence of autophagy and apoptosis, we used fluorescence microscopy and Western blot analysis. Results: The viability against hypoxia-reoxygenation injury in remifentanil preconditioning keratinocytes were increased, and these cells were showed stimulated expression of autophagy 3-MA suppressed the induction of autophagy effectively and the protective effects on apoptosis. Atg5, Beclin-1, LC3-II and p62 were elevated in RPC/HR group. But they were decreased when autophagy was suppressed by 3-MA. Conclusions: Remifentanil preconditioning showed the protective effect in human keratinocytes, and we concluded that autophagy may take the major role in the recovery of wound from hypoxia-reoxygenation injury. We suggest that further research is needed about the cell protective effects of autophagy.

Effects of Nicotine on the Expression of Cell Cycle Regulatory Proteins of Human Gingival Fibroblasts (니코틴이 치은섬유아세포의 세포주기 조절 단백질 발현에 미치는 영향)

  • Kim, Tak;Kim, Jae-ho;Pi, Sung-Hee;Kim, Eun-Cheol;You, Yong-Ouk;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.31 no.3
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    • pp.597-610
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    • 2001
  • Normal gingival fibroblasts functioning is fundamental for the maintenance of periodontal connective tissue as well as wound healing. Nicotine have been found to affect DNA synthesis and cell proliferation, which appear to depend on the type of cells. This in vitro study was done to determine the effects of nicotine, a major component of tobacco, on cell proliferation, viability, activity, cell cycle distribution, and expression of cell cycle regulatory proteins in human gingival fibroblasts. Nicotine has been tested for 2 days or 4 days in 5 different concentrations; $0.1{\mu}g/ml$; $1{\mu}g/ml$; $10{\mu}g/ml$; $100{\mu}g/ml$; $1000{\mu}g/ml$. To assess cell proliferation and viability, viable and non-viable cells were counted by hemocytometer; to evaluate cellular activity, MTT assay was employed; to analyze cell cycle distribution, fluorescent propidium iodide-DNA complex were measured using fluorocytometer; to determine the expression of cell cycle regulatory proteins, western blot analysis was performed. After 2 days and 4 days incubation respectively, at concentrations of $1{\mu}g/ml$ - $1000{\mu}g/ml$, nicotine significantly inhibited proliferation comparing to non-supplemented controls. The cell viability was significantly decreased after 2 days and 4 days at concentrations of $1{\mu}g/ml$ - $1000{\mu}g/ml$ and at $10{\mu}g/ml$ - $1000{\mu}g/ml$ respectively. After 2 days and 4 days, the cellular activity was significantly decreased at concentrations of $10{\mu}g/ml$ - $1000{\mu}g/ml$. Treatment with $100{\mu}g/ml$ nicotine for 48 hours caused an increase in the proportion of G1-phase cells (from 46.41% to 53.46%) and a decrease in the proportion of S-phase cells (from 17.80% to 14.27%). The levels of cyclin $D_1$ and CDK 4 proteins in nicotine-treated fibroblasts were lower than that of controls, whereas the levels of p16 and pRB were higher than that of controls. These results suggest that the decrease of cell proliferation and lengthened Gap phases (G1) by nicotine may due to the increased expression of p16 and pRB as well as decreased expression of cyclin $D_1$ and CDK 4 in human gingival fibroblasts.

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