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Expression of EGFP in Bovine Embryos after Intracytoplasmic Sperm Injection using Spermatozoa Co-cultured with Exogenous DNA (소 난자에 있어서 외래유전자가 도입된 정자의 직접 주입에 의한 EGFP 의 발현)

  • Lee, . H. C.;S. J. Uhm;S. Y. Ann;H. J. Chung;Park, H. D.;Lee, H. T.;K. S. Chung
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.439-449
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    • 2000
  • This study was to investigate the expression of transgene after co-injection of spermatozoon and EGFP gene into mature oocytes in cattle. From frozen semen, spermatozoa were treated by DTT with 0.03% Tween-20, freezing and thawing or 0.02% Triton X-100 to disrupt their plasma membranes. The sperm injected oocytes were co-cultured with bovine oviduct epithelial cells in CRlaa, and expression of EGFP in embryos were observbed under epifluorescent microscope. Two pronuclei were formed in oocytes injected with sperm treated by DTT (44.6%), DTT-Tween-20 (48.4%), DTT-freezing (44.4%) and DTT-Triton X-100 (42.9%). Cleavage and blastocyst formation rates of bovine oocytes which injected with sperm treated by DTT, DTT-Tween-20, DTT-freezing, and DTT-Triton X-100 were 49.1, 58.5, 43.9, and 48.4% and 10.2, 13.0, 6.8, and 6.5%, respectively. Although the most of embryos were showing mosaicism, embryos expressing EGFP gene were observed in all treated groups. Therefore, these results indicate that membrane disrupted sperm could interact with exogenous DNA, and that this procedure may be useful to introduce foreign gene into bovine oocytes.

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Effect of Leptin on the Expression of Lipopolysaccharide-Induced Chemokine KC mRNA in the Mouse Peritoneal Macrophages

  • Lee, Dong-Eun;Kim, Hyo-Young;Song, In-Hwan;Kim, Sung-Kwang;Seul, Jung-Hyun;Kim, Hee-Sun
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.722-729
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    • 2004
  • Leptin is an adipocyte-secreted hormone and its plasma levels correlate with total body fat mass, however, it also plays a regulatory role in immunity, inflammation, and hematopoiesis. Chemokine is known as a chemoattractant cytokine in inflammatory reaction, but its role in leptin reaction has not been well studied. In this study, the direct effect of leptin on the expression of chemokine mRNAs and lipopolysaccharide (LPS)-induced chemokine KC mRNA in mouse peritoneal macrophages was investigated. Leptin did not induce the expression of lymphotactin, RANTES, eotaxin, MIP-1$\beta$, MIP-1$\alpha$, MIP-2, MCP-1, IP-10, TCA-3, and KC mRNA in mouse peritoneal macrophages, and had no direct effect on the expression of these LPS-induced chemokine mRNAs except KC mRNA. The synergistic effect of leptin on the expression of LPS-induced KC mRNA occurred late in the time course of response to LPS. The increased expressions of Ob-Rb mRNA and leptin receptor protein were detected during the LPS treatment. Leptin produced a substantial increase in the stability of the LPS-induced KC mRNA, and the synergistic effect of leptin on LPS-induced KC mRNA expression was further augmented by cycloheximide (CHX). Pyrrolidine dithiocarbamate (PDTC) did not block the synergistic effect of leptin on LPS-induced KC mRNA expression in mouse peritoneal macrophages. These data suggest that although leptin has no direct effect on the expression of lymphotactin, RANTES, eotaxin, MIP-1$\beta$, MIP-1$\alpha$, MIP-2, MCP-1, IP-10, TCA-3, and KC mRNA in mouse peritoneal macrophages, the synergistic effect of leptin on the expression of LPS-induced KC mRNA has the possibility that LPS might induce the expression of the Ob-Rb receptor or an unknown gene(s) that sensitizes macrophages to the synergistic function of leptin. Therefore, further studies are necessary to examine leptin as a regulatory factor of chemokine production.

Korean solar salts reduce obesity and alter its related markers in diet-induced obese mice

  • Ju, Jaehyun;Song, Jia-Le;Park, Eui-Seong;Do, Myoung-Sool;Park, Kun-Young
    • Nutrition Research and Practice
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    • v.10 no.6
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    • pp.629-634
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    • 2016
  • BACKGROUND/OBJECTIVE: The aim of this experiments was to show anti-obesity effects of Korean solar salt from different salt fields in diet-induced obese mice. MATERIALS/METHODS: Diet-induced obesity (DIO) was induced by a high-fat diet (HFD; 45% cal from fat) in C57BL/6J mice for eight weeks. The mice were fed with the designated diets (chow diet for Normal, HFD for Control, 0.47%-salt-mixed HFD for purified salt (PS), Guerande solar salt from France (SS-G), solar salt from Y salt field (SS-Y), solar salts from T salt field (SS-T) and S salt field (SS-S)) for another eight weeks. We checked body weight, food efficiency ratio (FER) and tissue weights (liver and epididymal adipose tissue (EAT)), and observed serum concentrations of triacylglycerol (TG), total cholesterol (TC), leptin and insulin. We also evaluated gene expressions of adipogenic / lipogenic mRNAs of $C/EBP{\alpha}$, $PPAR{\gamma}$ and FAS and beta-oxidation-related factors ($PPAR{\alpha}$ and CPT-1) in liver and EAT. The mineral composition of salt samples were analyzed using inductively coupled plasma optical emission spectrometry (ICP-OES). RESULTS: SS-T and SS-S significantly reduced body weight gain, FER, and weight of EAT compared to control and other samples (P < 0.05). SS-T and SS-S also significantly decreased serum levels of TG, TC, leptin and insulin (P < 0.05). SS-T and SS-S suppressed expressions of adipogenic / lipogenic mRNAs in liver and EAT, while promoting expression of beta-oxidation-related factors. The lowest sodium concentration was observed in SS-T ($30.30{\pm}0.59%$), and the lowest sodium-to-potassium (Na/K) ratio was found in SS-S (17.81). CONCLUSIONS: Our study shows that well-processed Korean solar salt may have anti-obesity effects in vivo, probably owing to its differences in mineral composition and other components, presumably resulting from the manufacturing processes. Further research is needed into the mechanism and to explore optimal manufacturing processes.

A case of Smith-Lemli-Opitz syndrome diagnosed by identification of mutations in the 7-dehydrocholesterol reductase (DHCR7) gene (7-dehydrocholesterol reductase (DHCR7) 변이로 진단된 Smith-Lemli-Opitz 증후군 1예)

  • Park, Mee Rim;Ko, Jung Min;Cheon, Chong-Keun;Kim, Gu-Hwan;Yoo, Han-Wook
    • Clinical and Experimental Pediatrics
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    • v.51 no.11
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    • pp.1236-1240
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    • 2008
  • Smith-Lemli-Opitz syndrome (SLOS) is a rare, autosomal recessive disease caused by an inborn error in cholesterol synthesis. Patients with this disease suffer from multiple malformations due to reduced activity of 7-dehydrocholesterol reductase (DHCR7), which increases 7-dehydrocholesterol (7DHC) and 8-dehydrocholesterol (8DHC) concentrations and decreases cholesterol concentration in body fluids and tissue. The SLOS phenotypic spectrum ranges from a mild disorder with behavioral and learning problems to a lethal disease characterized by multiple malformations. Here, we describe a newborn male with ambiguous genitalia who was diagnosed to have type II SLOS during the neonatal period. A clinical examination revealed low levels of unconjugated estriol in the maternal serum, and a variety of fetal ultrasound anomalies, including prenatal growth retardation. After birth, the infant was diagnosed to have congenital heart disease (Tetralogy of Fallot with severe pulmonary artery stenosis), cleft lip and palate, micrognathia, postaxial polydactyly, ambiguous genitalia, and cataracts. Clinical investigation revealed extremely low plasma cholesterol levels and the presence of mutation (homozygote of p.Arg352Gln) in the DHCR7 gene. The patient underwent palliative heart surgery (to widen the pulmonary artery) and received intravenous lipid supplementation. Cholesterol levels increased slightly, but not to normal values. The patient died from cardiopulmonary failure and sepsis 72 days after birth. This report provides the first description of a Korean patient with SLOS confirmed by verification of DHCR7 gene mutation and illustrates the need for early recognition and appropriate diagnosis of this disease.

Involvement of Kupffer Cell in $CCl_4$ induced Liver Injury: The Role of Calcium (사염화 탄소에 의한 간손상에 있어 Kupffer cell 칼슘의 역할)

  • Yang, Mie-Rha
    • The Korean Journal of Pharmacology
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    • v.32 no.1
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    • pp.75-82
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    • 1996
  • The hypothesis that calcium provoke $O_2^-$ formation by Kupffer cells and may contribute to carbon tetrachloride $(CCl_4)$ induced liver injury was studied in SD rats. In $CCl_4-treated$ animals, hepatic malonaldehyde (nmole/gm liver) and plasma ALT (IU/ml) levels elevated significantly from $119.63{\pm}13.00$ to $268.97{\pm}14.82$ and from $17.3{\pm}0.18$ to $806.08{\pm}37.63$, respectively, compared to those in controls. Activation of Kupffer cells with high dose of retinol (250,000 IU/kg/day, po, for 7 day) significantly enhanced ALT levels, while inactivation of Kupffer cells with gadolinium chloride (7.5 mg/kg/day, ip, for 2 day) attenuated the increase of serum ALT level following $CCl_4$ treatment. Diltiazem (10 mg/kg/day, ip for 2 day) given in combination with retinol led to a marked decrease in ALT levels compare to the level in rats treated only with retinol against $CCl_4$ treatment. In order to determine any alterations in cytochrome P450 activities, the P450 content and the CYP2E1 activity were measured and all $CCl_4-treated$ rats showed significantly lower levels compared to those in controls and vehicle-treated animals. There were significant increases in glutathione peroxidase in all $CCl_4-treated$ rats except diltiazem treated groups. No difference was found among untreated and vehicle-treated rats. It is concluded that Kupffer cells contribute to $CCl_4-induced$ liver injury and that calcium antagonist attenuated the increased $CCl_4-induced$ liver injury due to activation of Kupffer cells.

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Effects of Cyclobuxine D on Carrageenin-induced Pleurisy and Croton Oil-induced Granuloma Pouch in Rats (흰쥐의 Carrageenin 유발 늑막염과 Croton oil 유발 육아종양에 미치는 Cyclobuxine D의 영향)

  • Lee, Jong-Hwoa;Park, Young-Hyun;Cho, Byung-Heon;Kim, Yu-Jae;Kim, Jong-Bae;Kim, Chung-Mok;Kim, Chun-Sook;Cha, Young-Deog;Kim, Young-Suk
    • The Korean Journal of Pharmacology
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    • v.23 no.2
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    • pp.151-157
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    • 1987
  • Cyclobuxine D, a steroidal alkaloid, was extracted from Buxus microphylla var. koreana Nakai. The effects of cyclobuxine D on carrageenin-induced pleurisy and croton oil-induced granuloma pouch in rats was investigated and compared with those of aspirin, hydrocortisone ana dexamethasone. Intrapleural injection of 2% carrageenin caused the accumulation of exudate. The rate of plasma exudation, measured by the exuded dye amounts for 20 min in the pleural cavity after intravenous injection of pontamine sky blue, showed a peak at 5 hr. Cyclobuxine D (5, 20 and 50 mg/kg, i.p.) suppressed dose-dependently the accumulation of the pleural exudate and the exudation of dye. Among several methods used for screening and evaluation anti-inflammatory agents, granuloma pouch technic introduced by Hans Selye (Hans seyle, 1953) is considered as a simple and reliable method. An air pocket was produced in the subcutaneous tissue of the interscapular region by injection of 1 ml of 1% croton oil as irritant. Inflammatory exudate accumulated in the pouch during the succeding 14 days. Cyclobuxine D (5 and 20 mg/kg) decreased fluid volume in pouch and weight of pouch wall in granulomatous inflammation.

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Giant Neurofibroma on Both Buttocks (양측 엉덩이의 거대 신경섬유종)

  • Kim, Ji Hoon;Burm, Jin Sik;Kim, Yang Woo;Kang, So Ra;Kim, Hyoung Kyoung
    • Archives of Plastic Surgery
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    • v.36 no.4
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    • pp.512-515
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    • 2009
  • Purpose: Neurofibromatosis(NF) is an autosomal - dominant systemic disease. Up to fifty percent of patients with NF are reported to have concomitant vascular abnormalities. In the resection of a larger NF, the risk of uncontrolled hemorrhage is much higher due to the difficulty of hemostasis of large vessels within the tumor. We ligated the base of the giant NF with a simple loop - shaped ligation before removal of the giant NF in both buttocks. And then we could successfully reduce the amount of hemorrhage during the operation. Methods: A 46 - year - old female patient presented for giant masses of both gluteal area, which has been growing slowly for the last ten years. Each mass was about $30{\times}20cm$ in size. After designing the elliptical resection margin, we tightened the tumor base by using continuous loop - shaped suture ligation(weaving the thread up and down in a loop - shaped pattern, leaving a space of 2 cm between each loop) with a straight needle and prolene 2 - 0. After skin incision, we proceeded the dissection toward the central and inferior side of the mass obliquely while we avoided breaking large vascular sinuses. We resected the tumor in a wedged - shape. Subcutaneous tissue was sutured layer by layer and skin was closed by vertical mattress and interrupted suture. The loop - shaped ligation of the base was removed and compressive dressing was done with gauzes and elastic bandages. Results: Postoperative complications such as infection, hemorrhage, hematoma, and dehiscense did not occur. Perioperatively the patient was sufficiently transfused with five units of blood and two units of fresh frozen plasma. During the subsequent 1 year follow - up, the functional and cosmetic result was excellent. Conclusion: A continuous loop - shaped suture ligation procedure along the base of the giant NF effectively reduced the amount of hemorrhage during the operation, made dissection and ligation of vessels easily and quickly, and shorten the operating time and postoperative recovery time.

Antioxidant Activities of Glycyrrhizin and its Effect on Renal Expression of Na,K-ATPase in Gentamicin-induced Acute Renal Failure Rats (Glycyrrhizin의 항산화 활성 및 Gentamincin 유도 급성 신부전 백서 신장의 Na,K-ATPase 발현에 미치는 영향)

  • Sohn Eun Jin;Kang Dae Gill;Lee An Sook;Lee Yun Mi;Yin Ming Hao;Yeum Kee Bok;Noh Suk Yun;Lee Ho Sub
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.542-548
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    • 2003
  • The present study was aimed to investigate whether glycyrrhizin, which is the major component of Glycyrrhiza uralensis, has an antioxidant effect and regulatory effect on Na,K-ATPase in gentamicin-induced acute renal failure (ARF) rats . It is well known that reactive oxygen species (ROS), such as superoxide anion and hydroxyl radical, are main pathophysiological factor in gentamicin-induced ARF. Glycyrrhizin showed potent in vitro antioxidant activity, especially superoxide scavenging activity, in a dose-dependent manner. Plasma lipid peroxide level was restored to normal level by oral administration of glycyrrhizin (200 mg/kg) in the gentamicin-induced ARF rats. The expression of Na,K-ATPase α1 subunit was restored in the gentamicin-induced ARF rats by administration of glycyrrhizin, whereas β1 subunit was not restored. The renal functional parameters including urine volume, cleatinine clearance, urine osmolality, solute-free water reabroption were also partially restored in gentamicin-ARF rats by administration of glycyrrhizin. Taken together, the amelioration of renal functions and the expression of sodium pump by administration of glycyrrhizin in the gentamicin-induced ARF was appear to be mediated by the scavenging of ROS.

Anti-inflammatory action by Gamisangryosamul-tang and The effect of Ziyang-Go on atopic dermatitis-like lesion and pruritus in NC/Nga mice (기미생료사물탕의 항염증효과와 지양고의 아토피피부염 손상 및 지양 효과에 미치는 영향)

  • Kim Jeong Jin;Yang Sung Wan;Son Nak Won;Ahn Kyoo Seok
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.428-435
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    • 2003
  • This research was performed to examine an anti-inflammatory effects of Gamisangryosamul-Tang(GSS) and anti-pruritus effects of Ziyang-Go(Salve). This study was processed by three experiments; Experiment 1: Inhibitory activity of GSS extract on the degranulation of mast cell and histamine release in plasma induced by compound 48/80 i.p, injection after the pretreatment of GSS extract i.p, injection in Sprague-Dawley rats, Experiment 2: Anti-inflammatory effect of GSS extract on macropharge raw 264,7 cells treated by LPS 250 ppm (before 2 hours). Experiment 3: Measurement of passive cutaneous anaphylaxis and atopic dermatitis using NC/Nga mice, GSS extract inhibited histamine release by 70% compared to compound 48/80 treated control group and histologically significantly reduced (P<0,01) the degranulation of mast cell in SD rats. In GSS extract treated group, the expression of TNF-α in macropharge cell showed the remarkable inhibitory effect about 62% (P<0,01) compared to LPS treated control group. The expression of IL-6 appeared more effective by 46% than the LPS treated control group and by 6% compared to hydrocortison treated group, Comparing with steroid (0.05% prednisolon) ointment, Ziyan-Go treated group showed the significant(30%) recovery on skin response index in atopic dermatis like anaphylaxis mice(NC/Nga), Finally, in scratching behavioral tests of NC/Nga mice for three weeks, Ziyang-Go treated group significantly (P<0.05) suppressed the pruritus on the face, neck, ears and dorsal skin than inbred NC/Nga mice. However, the change of IgE and IFN-γ from the spleen cell of NC/Nga mice was not significantly different between the oral intake of GSS extract group and of saline intaked control group. Summary and Conclusion: This study demons trates that Ziyang-go have the equal anti-pruritus effect to steroid ointment and GSS extract have the notable immunologic activity on inflammatory in vivo and in vitro model. Advanced experiment of this study will be required for more reliable information about the correlation between the lymphokine (i.e. IgE) and the anti-allergic effects of GSS.

Protective effect of Ganopoly and Ganopoly/C+ on nephrotoxicity induced by cisplatin in rats (Cisplatin의 신장독성에 대한 영지추출물 복합제제의 보호효과)

  • Kim Dae Geun;Kim Kun Jung;Ju Sung Min;Kim Yong Ik;Choi Ho Seung;Keum Kyung Soo;Kim Won Sin;Gao Yiu Ai;Jeon Byung Hun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.316-325
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    • 2003
  • In this paper, the effect of Ganopoly(extracts of Ganoderma lucidum) and Ganopoly/C+(70% Ganopoly + 30% chitosan) on cisplastin-induced nephrotoxicity was investigated in Sprague-Dawley rats. A single dose of cisplastin(5 ㎎/㎏) kg) was administered intraperitoneally after pretreatment of saline, Ganopoly and Ganopoly/C+ for 7 days. The nephrotoxicity and renal function were manifestated by the changes of body weight, blood pressure, biochemical changes and solute in urine and plasma. After the treatment of CDDP(cis-dichlorodiamineplatinum), a significant elevation of kidney weight, serum urea, cretinine, urine volume for 24 hours, urine magnesium, and a severe or significant decrease in body weight, blood pressure, creatinine clearance, urine osmolarity, serum albumin, etc. The nephrotoxicity was further confirmed by a significant decrease in glutathione S-transferase(GSH) in urine and kidney homogenate, GSH, glutathione peroxidase(GSH-Px) and catalase in kidney tissue. And also the lipid peroxidation was significantly increased in kidney homogenate. These signs of nephrotoxicity was ameliorated by the pretreatment and consecutive administration of Ganopoly and Ganopoly/C+ for 14 days after the Lp. injection of CDDP on 7th day after pretreatment of Ganopoly and Ganopoly/C+. The amelioration of nephrotoxicity was evidenced by significant reduction in serum urea and creatinine concentration, and improvement of other index of renal function. And The activity of antioxidant enzymes were partially recovered in kidney tissue of rats treated by CDDP and the administration of Ganopoly and Ganopoly/C+. These results indicate the cispastin induced nephrotoxicity is due to an impairment of tubular reabsorption systems enhanced by necrosis of proximal tubule, and the Ganopoly and Ganopoly/C+ has a partial protective effect on nephrotoxicity induced by CDDP. The polysacchride of Ganoderma lucidum may improve the therapeutic index of nephrotoxicity induced by CDDP. However, it is needed to elucidate the mechanism for confirming the therapeutic effect.