• Title/Summary/Keyword: CK1

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Acute Toxicity of pCK-VEGF in Rata and Mice (Vasular Endothelial Growth Factor 165 (VEGF165) 발현 벡터 (pCK-VEGF)의 랫드와 마우스에서의 급성독성)

  • 김선영;이영주;조홍찬;박은진;안병옥;김덕경
    • Toxicological Research
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    • v.16 no.1
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    • pp.67-71
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    • 2000
  • It has been demonstrated that the injection of naked DNA expressing vascular endothelial growth factor 165(VEGFl165) to the affected area can provide significant therapeutic effects on peripherol artery occlusive diseases. Success with this type of gene therapy highly depends on the quality of the vector delivering the therapeutic gene, especially in terms of the level and duration of gene expression in the localized area. We have recently developed a vector expressing VEGF165(pCK-VEGF) for the treatment of peripheral artery occulsive diseases and demonstrated high level expression of VEGF165 in mouse skeletal muscle. This study was designed to assess the acute toxicity of intramuscularly injected pCK-VEGF in BALB/c mice and Sprague-Dawely rats. There was no evidence of any changes in clinical signs, body weights, or gross pathological signs. We estimate LD50 values of pCK-VEGF higher than 50mg/kg in mice and 20 mg/kg in rats by intramuscular injection.

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Ginsenoside compound K protects human umbilical vein endothelial cells against oxidized low-density lipoprotein-induced injury via inhibition of nuclear factor-κB, p38, and JNK MAPK pathways

  • Lu, Shan;Luo, Yun;Zhou, Ping;Yang, Ke;Sun, Guibo;Sun, Xiaobo
    • Journal of Ginseng Research
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    • v.43 no.1
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    • pp.95-104
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    • 2019
  • Background: Oxidized low-density lipoprotein (ox-LDL) causes vascular endothelial cell inflammatory response and apoptosis and plays an important role in the development and progression of atherosclerosis. Ginsenoside compound K (CK), a metabolite produced by the hydrolysis of ginsenoside Rb1, possesses strong anti-inflammatory effects. However, whether or not CK protects ox-LDL-damaged endothelial cells and the potential mechanisms have not been elucidated. Methods: In our study, cell viability was tested using a 3-(4, 5-dimethylthiazol-2yl-)-2,5-diphenyl tetrazolium bromide (MTT) assay. Expression levels of interleukin-6, monocyte chemoattractant protein-1, tumor necrosis factor-${\alpha}$, intercellular adhesion molecule-1, and vascular cell adhesion molecule-1 were determined by enzyme-linked immunosorbent assay and Western blotting. Mitochondrial membrane potential (${\Delta}{\Psi}m$) was detected using JC-1. The cell apoptotic percentage was measured by the Annexin V/ propidium iodide (PI) assay, lactate dehydrogenase, and caspase-3 expression. Apoptosis-related proteins, nuclear factor $(NF)-{\kappa}B$, and mitogen-activated protein kinases (MAPK) signaling pathways protein expression were quantified by Western blotting. Results: Our results demonstrated that CK could ameliorate ox-LDL-induced human umbilical vein endothelial cells (HUVECs) inflammation and apoptosis, $NF-{\kappa}B$ nuclear translocation, and the phosphorylation of p38 and c-Jun N-terminal kinase (JNK). Moreover, anisomycin, an activator of p38 and JNK, significantly abolished the anti-apoptotic effects of CK. Conclusion: These results demonstrate that CK prevents ox-LDL-induced HUVECs inflammation and apoptosis through inhibiting the $NF-{\kappa}B$, p38, and JNK MAPK signaling pathways. Thus, CK is a candidate drug for atherosclerosis treatment.

Elevated Serum Creatine Kinase Level in Rotavirus Gastroenteritis (Rotavirus 위장관염에서 혈청 Creatine Kinase의 증가)

  • Bae, Won Tae;Kim, Jae Hui;Park, Eun Sil;Seo, Ji Hyun;Lim, Jae Young;Park, Chan Hoo;Woo, Hyang Ok;Youn, Hee Shang
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.8 no.2
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    • pp.117-121
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    • 2005
  • Purpose: Elevated AST/ALT level in rotavirus gastroenteritis have been reported. We found elevated creatine kinase (CK) in those children. This study was carried out to define the relationship of major clinical presentation, laboratory findings, and level of creatine kinase. Methods: A retrospective analysis was made for thirty patients who were diagnosed rotavirus gastroenteritis from Jan 2001 to Mar 2005 in Department of Pediatrics, Gyeongsang National University Hospital. Serum creatine kinase was checked for the evaluation of high aminotransferase level among the patients. Results: Fourteen patients (8 males, 6 females) with high creatine kinase activity were included in this study. The mean age was $1.46{\pm}1.24year$ of age. The mean level of AST, ALT and CK were $127.5{\pm}136.2IU/L$, $126.1{\pm}154.3IU/L$, and $542.8{\pm}624.6IU/L$, respectively. Electrophoresis of CK isoenzyme was performed in four of them and the results revealed elevated CK-MM fraction (96~100%). Three of them revealed high serum creatine kinase acitivity (>1,000 IU/L) without acute renal failure and other symptom. However, none of them had muscular pain or trauma history. Elevated creatine kinase activity did not correlate with clinical implications (age, sex, vomiting, diarrhea, fever, symptom of URI, degree of dehydration or seizure) or laboratory findings. Conclusion: In this study, we found that serum creatine kinase acitivity also was elevated in infants with rotavirus gastroenteritis. This data support muscular damage due to rotavirus, but could not prove the mechanism of increased serum creatine kinase activity.

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Changes of Plasma Creatinine Kinase-BB after Total Circulatory Arrest (총순환정지후 혈중 크레아티닌 카이네이즈 BB의 변화에 관한 연구)

  • 이석재;김용진;김오곤
    • Journal of Chest Surgery
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    • v.31 no.10
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    • pp.945-951
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    • 1998
  • Background: Although profound hypothermia with total circulatory arrest(TCA) is a valuable maneuver in cardiac surgery, its applications have been limited due to serious complications, especially cerebral damage. In this study, the possible role of creatinine kinase-BB(CK-BB), an index enzyme of ischemic cerebral damage, was assayed as a parameter for the assessment of the cerebral complications after TCA. Hemoglobin(Hb), ionized calcium(Ca++), and blood glucose levels were also assessed as clinical parameters involved in cerebral damage. Materials and methods: Among patients with congenital heart disease, 18 patients who had been operated on with TCA were randomly selected and divided into two groups: 6 with acyanotic and 12 with cyanotic heart disease. Arterial blood from each patient was collected before and after TCA at scheduled times(15 min., 30 min, 1, 2, 4, 8, and 12hr). The levels of CK-BB, Hb, Ca++, and blood glucose were assessed in each sample. Results: As a whole, correlation between CK-BB level and blood sampling time after TCA was not statistically significant. Also, the difference in the level of CK-BB after TCA was not significant between the acyanotic and cyanotic groups. The levels of Hb and CK-BB correlated significantly. Conclusions: The results, which showed no correlation between the alterations in CK-BB level and the TCA duration, suggest that the single assay of the CK-BB level is not a representative measurement for the assessment of cerebral damage after TCA. Also, the cyanotic congenital heart disease group is not more vulnerable to cerebral damage induced by TCA.

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Expression of a Bacillus subtilis Endoglucanase in Protease-Deficient Bacillus subtilis Strains

  • Yang, Mi-Jeong;Jung, Sun-Hwa;Shin, Eun-Sun;Kim, Jung-Ho;Yun, Han-Dae;Wong, Sui-Lam;Kim, Ho-On
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.430-434
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    • 2004
  • Three extracellular protease-deficient Bacillus subtilis strains were transformed with the plasmid pCK98 containing the endo-$\beta$-1,4-glucanase (Eng) gene of B. subtilis BSE616. The three transformants, B. subtilis DB104 (pCK98), WB600 (pCK98) and WB700 (pCK98), produced the same high level of enzyme activity and showed similar patterns of cell growth and enzyme production. When B. subtilis DB 104 (pCK98), a two-extracellular protease deficient strain, was cultured for 22 h, almost all the secreted enzyme was found to be in the completely cleaved form by both activity staining and Western blotting studies. B. subtilis WB600 (pCK98), a six-extracellular protease-deficient strain, produced a partially cleaved form in addition to the intact form of the enzyme, although the degree of internal cleavage of the enzyme was greatly reduced. With B. subtilis WB700 (pCK98), a seven-extracellular protease-deficient strain, almost all the enzyme was produced as the intact uncleaved form. This study illustrates that a role of the V pr protease is to degrade foreign proteins produced in B. subtilis and WB700 is a suitable expression system for producing the intact form of the Eng and other foreign proteins that may lose at least part of their efficacy due to internal proteolytic cleavage.

Cloning and Expression in E. coli of the HOPDA Hydrolase Gene from Pseudomonas sp. P20

  • Lim, Jong Chul;Chae, Jong Chan;Kim Youngsoo;Kim, Hyong Bai;Kim Chi Kyung
    • Journal of Microbiology
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    • v.34 no.4
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    • pp.349-354
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    • 1996
  • Pseudomonas sp. P20 is a natural isolate which is capable of degrading biphenyl and 4-chlorobiphenyl. From a clone of pCK1022 harboring pcbCD genes of Pseudomonas sp P20, a pcbD gene encoding 2-hydroxy-6-oxo-6-phenylhexa-2, 4-dienoic acid (HOPDA) hydrolase was subcloned in Escherichia coli XL-1-Blue by using pBluescript SK(+) vector. The 2.8-kb HindII fragment harboring the pcbD gene cloned in pCK 1024 had a single site for each of XhoI, SalI, BstXI, and XbaI restriction enzymes. Escherichia coli CK1024 had a single site for each of XhoI, SalI, BstXI, and XbaI restriction enzymes. Escherichia coli CK1024 carrying pCK0124 degraded HOPDA to benzoate and 2-hydroxypenta-2, 4-dienoate by HOPDA hydrolase encoded by pcbD gene as effectively as E coli CK 1022 HARBORING pcbCD genes.

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[ $Kienb\"{o}ck's$ ] Disease Treated With Vascular Loop Graft (Vascular loop graft 술식을 이용한 $Kienb\"{o}ck$씨 질환의 치료)

  • Han, Chung-Soo;Chung, Duke-Whan;Park, Bo-Yeon;Nam, Gi-Un;Han, Hyun-Soo
    • Archives of Reconstructive Microsurgery
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    • v.2 no.1
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    • pp.13-19
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    • 1993
  • Although the etiology of $Kienb\"{o}ck's$ disease is clearly related to avascular changes in the lunate, but the actual cause leading to this vascular impairment has remained elusive. Therefore, a great many different surgical procedures have been proposed for the correction of the multiple factors leading to lunate collapse or for the treatment of the lunatomalacia. The treatment modalities includes lunate excision, intercarpal arthrodesis, lunate implant resection arthroplasty, joint levelling operation(e.g ulnar lengthening & radial shortening), pronater quadratus pedicle graft and vascular loop graft. In the period from Jan. 1981 to Dec. 1992, we performed operative treatment in 19 cases of $Kienb\"{o}ck's$ disease. Among them, 6 cases were treated with vascular loop graft. We analysed all patients who were treated with vascular loop graft after followed up of 4 year 6 months, on an average(range from 1 year to 8 year 10 months). The results analysed are as follows, 1. All cases were stage III according to Lichtman's classification. 2. Ulnar variance was -1.5(range$-2{\sim}0$), on an average. 3. The average age of patients were 37.7years old(range 31-41). 4. Postoperatively, there were considerable restoration of range of motion and complete relief of pain in all cases, but continued decrease of grip power in one case. 5. Decreased sclerosis, loss of fragmentation and new bone formation were appeared in the last follow up film, in all cases. The vascular loop graft considered as a useful method for the treatment of the $Kienb\"{o}ck's$ disease.

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Reciprocal regulation of SIRT1 and AMPK by Ginsenoside compound K impedes the conversion from plasma cells to mitigate for podocyte injury in MRL/lpr mice in a B cell-specific manner

  • Ziyu Song;Meng Jin;Shenglong Wang;Yanzuo Wu;Qi Huang;Wangda Xu;Yongsheng Fan;Fengyuan Tian
    • Journal of Ginseng Research
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    • v.48 no.2
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    • pp.190-201
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    • 2024
  • Background: Deposition of immune complexes drives podocyte injury acting in the initial phase of lupus nephritis (LN), a process mediated by B cell involvement. Accordingly, targeting B cell subsets represents a potential therapeutic approach for LN. Ginsenoside compound K (CK), a bioavailable component of ginseng, possesses nephritis benefits in lupus-prone mice; however, the underlying mechanisms involving B cell subpopulations remain elusive. Methods: Female MRL/lpr mice were administered CK (40 mg/kg) intragastrically for 10 weeks, followed by measurements of anti-dsDNA antibodies, inflammatory chemokines, and metabolite profiles on renal samples. Podocyte function and ultrastructure were detected. Publicly available single-cell RNA sequencing data and flow cytometry analysis were employed to investigate B cell subpopulations. Metabolomics analysis was adopted. SIRT1 and AMPK expression were analyzed by immunoblotting and immunofluorescence assays. Results: CK reduced proteinuria and protected podocyte ultrastructure in MRL/lpr mice by suppressing circulating anti-dsDNA antibodies and mitigating systemic inflammation. It activated B cell-specific SIRT1 and AMPK with Rhamnose accumulation, hindering the conversion of renal B cells into plasma cells. This cascade facilitated the resolution of local renal inflammation. CK facilitated the clearance of deposited immune complexes, thus reinstating podocyte morphology and mobility by normalizing the expression of nephrin and SYNPO. Conclusions: Our study reveals the synergistic interplay between SIRT1 and AMPK, orchestrating the restoration of renal B cell subsets. This process effectively mitigates immune complex deposition and preserves podocyte function. Accordingly, CK emerges as a promising therapeutic agent, potentially alleviating the hyperactivity of renal B cell subsets during LN.

Regeneration of Bovine Mammary Gland in Immunodeficient Mice by Transplantation of Bovine Mammary Epithelial Cells Mixed with Matrigel

  • Park, Hyun Jung;Lee, Won Young;Jeong, Ha Yeon;Song, Hyuk
    • International Journal of Stem Cells
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    • v.9 no.2
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    • pp.186-191
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    • 2016
  • Background and Objectives: With the global demand for dairy protein for consumption growing annually, there has been increasing activity in the research field of dairy protein synthesis and production. From a manipulation perspective, it is more difficult to use live cattle for laboratory studies on the production of milk as well as of dairy protein such as casein, as compared with using laboratory animals like rodents. Therefore, we aimed to develop a mouse model of bovine mammary alveolar ducts for laboratory-scale studies. We studied the formation of the bovine mammary gland ductal structure by transplanting the MAC-T bovine alveolar cell line into mice. Methods and Results: MAC-T cells ($1{\times}10^7$) were suspended in Matrigel and injected into the dorsal tissue of 8-week-old male BALB/C nude mice. Histological analysis of tissue dissected from the MAC-T cell-transplanted mice after 6 weeks showed the typical morphology of the tubuloalveolar female gland, as well as glands made up of branching ducts that were surrounded by smooth muscle with small alveoli budding off the ducts. In addition, the epithelial markers CK14 and CK18 were expressed within the duct-like structure. Prolactin was detected in the duct interior in these CK14+ and CK18+ cells but not in the non-transplanted MAC-T cells. Conclusions: These results showed that duct-like tissue had been successfully formed after 6 weeks of transplantation of the CK14+ and CK18+ MAC-T cells into mice dorsal tissue. This mouse model will be a useful tool for further research on the bovine mammary gland.

Blood chemical research for the diagnosis of cardiac disease in ruminant (반추수 심장질병의 진단을 위한 혈액화학적 연구)

  • Kim, Duck-hwan;Lee, Kyo-young;Chang, Seuk-jin;Song, Kun-ho;Lee, Yoon-kyung;Yoon, Sang-bo
    • Korean Journal of Veterinary Research
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    • v.35 no.2
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    • pp.399-404
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    • 1995
  • In order to clarify the significance of serum CPK for the diagnosis of the cardiac disease in ruminant, the fluctuation of serum total CPK activities and CPK isoenzyme fractions was examined before and after operation in Korean native goats with artificially induced cardiac injury. In the change of serum total CPK activities, those of experimental group were changed with higher values than those of control group and significant increase was found on 2-week after operation(P<0.05). In the fluctuation of serum CPK isoenzyme fractions, significant increase of both CK1 and CK3 fractions was observed on 2-week after operation in experimental group(P<0.05), but significant difference was not found in CK2 fraction between experimental and control groups. From these findings, it is considered that taking note of increase of not only serum total CPK activities but also CK1 and CK3 fractions is important for the diagnosis of traumatic cardiac disease in ruminant.

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