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Bone Changes in Femoral Bone of Mice Using Calcein Labeling (Mice에서 Calcein 표지를 이용한 골 변화 관찰)

  • Shim, Moon-Jung
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.2
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    • pp.114-117
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    • 2016
  • In vivo labeling of bone with fluorochromes is a widely used method for assessment of bone formation and remodeling processes. In particular, calcein is used as a marker for identification of bone growth, which is indicated by a green color. Calcein green is a calcium chelator that adheres to regions of mineralizing bone thereby allowing localization of new bone. Bone formation and remodeling in vivo can be assessed by calcium-binding calcein labeling. In this study, changes in the femoral bone of a normal mouse model at both 4 and 8 weeks were evaluated using calcein labeling. Intense deposition of calcium in the bone was observed after application for 8 weeks. A mouse model is suitable for application in in vivo experiments using genetically modified mice, such as knock-out mice, however data regarding femoral cross sectional bone in young mice are limited. The current study confirmed calcein as a useful marker for identification of bone growth, which was indicated by a green color on photomicrographs. This methodological process may provide basic information for interpreting bone formation and regeneration to pharmacologic or genetic manipulation in mice.

Inhibitory Effect of a Decoction of Eucommiae ulmides OLIVER and Dipsacus asperoides C. Y. Cheng et T.M.Ai on Collagen II-induced Arthritis Mice (Collagen II-induced Arthritis 생쥐에 대한 두충(杜仲).속단(續斷) 배합약물의 관절염 억제 효과)

  • Lee, Boo-Kyun;Lee, Young-Cheol;Lee, Jang-Cheon
    • The Korea Journal of Herbology
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    • v.25 no.1
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    • pp.55-63
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    • 2010
  • Objectives : The object of this study was to verify the inhibitory effect of a decoction of Eucommiae ulmides OLIVER (EU) and Dipsacus asperoides C. Y. Cheng et T.M.Ai (DA) on Collagen II-induced Arthritis Mice (CIA mice). Methods : DBA/1OlaHsd mice were immunized with bovine type II collagen. Boostnig same collagen 21 days later, arthritis was induced and then administrated orally the extract of EU+DA (200 or 50 mg/kg) once a day for 4 weeks and compared with that of methotrexate (MTX, 0.3 mg/kg) as a positive control. Results : Administration of EU+DA suppressed the inflammatory progression of CIA mice and the results were 1. Arthritis index of CIA mice was decreased. 2. EU+DA decreased the production of TNF-$\alpha$, IL-6, IL-$1{\beta}$ in the serum of CIA mice. 3. EU+DA decreased the level of IgM. 4. EU+DAincreasaed $CD3^+$, $CD4^+$, $CD4^+$/CD25 but decreased $CD19^+$, $CD3^+/CD49b^+$(NKT), $CD3^-/CD49b^+$(NK), $B220^+/CD23^+$ in PBMC of CIA mice. 5. EU+DA decreased $CD3^+$, $CD4^+$, $CD3^+/CD69^+$ of paw joint in CIA mice. 6. EU+DA decreased subsynovial inflammation. Conclusions : This results demonstrated that extract of EU+DA suppressed the inflammatory progression of CIA mice and supported further studies are required to clarify a mechanism of therapeutic role.

Anti-diabetic Effect of Fermented Milk Containing Conjugated Linoleic Acid on Type II Diabetes Mellitus

  • Song, Kibbeum;Song, In-Bong;Gu, Hye-Jung;Na, Ji-Young;Kim, Sokho;Yang, Hee-Sun;Lee, Sang-Cheon;Huh, Chang-Ki;Kwon, Jungkee
    • Food Science of Animal Resources
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    • v.36 no.2
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    • pp.170-177
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    • 2016
  • Conjugated linoleic acid (CLA) is a group of positional and geometric isomers of conjugated dienoic derivatives of linoleic acid. CLA has been reported to be able to reduce body fat. In this study, we investigated the antidiabetic effect of fermented milk (FM) containing CLA on type II diabetes db/db mice. Mice were treated with 0.2% low FM, 0.6% high FM, or Glimepiride (GLM) for 6 wk. Our results revealed that the body weight and the levels of fasting blood glucose, serum insulin, and leptin were significantly decreased in FM fed mice compared to db/db mice. Oral glucose tolerance and insulin tolerance were significantly ameliorated in FM fed mice compared to db/db mice. Consistent with these results, the concentrations of serum total cholesterol, triglycerides, and LDL cholesterol were also significantly decreased in FM fed mice compared to db/db mice. However, the concentration of HDL cholesterol was significantly higher in FM fed mice compared to db/db mice. These results were similar to those of GLM, a commercial anti-diabetic drug. Therefore, our results suggest that FM has anti-diabetic effect as a functional food to treat type II diabetes mellitus.

Effect of Fasciculus Vascularis Luffae Herbal-Acupuncture at St36 on OVA-induced Asthma in Mice (사과락(絲瓜絡) 약침(藥鍼)의 천식억제(喘息抑制) 및 면역조절효과(免疫調節效果)에 대(對)한 실험적(實驗的) 연구(硏究))

  • Nam, Se-Hyeon;Hong, Kwon-Eui;Lee, Hyun
    • Journal of Acupuncture Research
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    • v.22 no.4
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    • pp.73-85
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    • 2005
  • Objectives : The purpose of this study is to confirm the suppression effect of Asthma and Immune response improvement of Fasciculus Vascularis Luffae Herbal-acupuncture into Chok-samni(St36) on ovalbumin-induced asthma in mice. Methods : C57BL/6 mice were sensitized and challenged with OVA(ovalbumin) for 12 weeks Two experimental groups were treated with different concentrations(1%, 0.1%) of FVL-HA at Chok-samni(St36) for the later 8 weeks(3times/week). Results : 1. Lung weigh of the mice group treated with FVL-HA decreased significantly compared with that of control group. 2. Total cells of lung, total Leukocytes and Eosinophils in BALF of the mice group treated with FVL-HA decreased significantly compared with those of control group. 3. Eosinophils in BALF of the mice group treated with FVL-HA in Photomicrographs decreased significantly compared with those of control group. 4. The concentration of IL-l3, IgE, IL-4 In serum and IL-4 in BALF of the mice group treated with FVL-HA decreased significantly compared with that of control group. 5. The number of Gr-1+/CB11b+, CD3-/CCR3+, CD4+, CD8+, CD3e+/CD69+, IgE+/B220+ cells in the lungs of the mice group treated with FVL-HA decreased significantly compared with those of control group. 6. The cytokine's manifestation of mRNA of the mice group treated with FVL-HA with RT-PCR decreased significantly compared with that of control group. Conclusion : These result suggests that Fasciculus Vascularis Luffae Herbal-acupuncture Choksamni(St36) in C57BL/6mice may be an effective part to OVA-induced asthma in C57BL/6 mice.

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Effects of Herbal-Acupuncture with Evodiae Fructus at KI10 on Osteoporosis in Ovariectomized Mice (음곡(陰谷) 오수유(吳茱萸) 약침(藥鍼)이 난소적출(卵巢摘出)생쥐의 골다공증(骨多孔症)에 미치는 영향(影響))

  • Choi, Sung-Hoon;Lee, Byung-Ryul;Yang, Gi-Young;Kim, Jae-Kue;Seo, Young-Suk;Yim, Yun-Kyoung
    • Korean Journal of Acupuncture
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    • v.27 no.2
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    • pp.217-242
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    • 2010
  • Objective : This study is aimed to evaluate the effects of Evodiae Fructus herbal-acupuncture (EF-HA) at KI10 on osteoporosis induced by ovariectomy in mice. Method : Mice underwent bilateral ovariectomy. After recovering, the ovariectomized (OVX) mice were treated by needle prick, saline injection, herbal acupuncture with Evodiae Fructus (EF-HA) at KI10 for 8 weeks. Result : 1. EF-HA at KI10 significantly inhibited the overgrowth of tibia in ovariectomized mice. 2. NP at KI10 significantly restored the tibial BMD (bone mineral density) in ovariectomized mice. 3. EF-HA at KI10 significantly restored the phosphorus and creatinine levels in ovariectomized mice serum. 4. EF-HA at KI10 significantly restored the tibial Ca and P levels in ovariectomized mice. 5. EF-HA at KI10 significantly reduced the tibial osteoclast-like cells in ovariectomized mice. 6. EF-HA at KI10 significantly inhibited the overgrowth of tibial GPL (growth plate length) in ovariectomized mice. Conclusion : EF-HA at KI10 has protective and therapeutic effect for osteoporosis in ovariectomized mice. Thus, it is suggested that EF-HA can be an useful therapeutics in clinical field after further researches.

IDH2 gene deficiency accelerates unilateral ureteral obstruction-induced kidney inflammation through oxidative stress and activation of macrophages

  • Kim, Jee In;Noh, Mi Ra;Yoon, Ga-Eun;Jang, Hee-Seong;Kong, Min Jung;Park, Kwon Moo
    • The Korean Journal of Physiology and Pharmacology
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    • v.25 no.2
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    • pp.139-146
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    • 2021
  • Mitochondrial NADP+-dependent isocitrate dehydrogenase 2 (IDH2) produces NADPH, which is known to inhibit mitochondrial oxidative stress. Ureteral obstruction induces kidney inflammation and fibrosis via oxidative stress. Here, we investigated the role and underlying mechanism of IDH2 in unilateral ureteral obstruction (UUO)-induced kidney inflammation using IDH2 gene deleted mice (IDH2-/-). Eight- to 10-week-old female IDH2-/- mice and wild type (IDH2+/+) littermates were subjected to UUO and kidneys were harvested 5 days after UUO. IDH2 was not detected in the kidneys of IDH2-/- mice, while UUO decreased IDH2 in IDH2+/+ mice. UUO increased the expressions of markers of oxidative stress in both IDH2+/+ and IDH2-/- mice, and these changes were greater in IDH2-/- mice compared to IDH2+/+ mice. Bone marrow-derived macrophages of IDH2-/- mice showed a more migrating phenotype with greater ruffle formation and Rac1 distribution than that of IDH2+/+ mice. Correspondently, UUO-induced infiltration of monocytes/macrophages was greater in IDH2-/- mice compared to IDH2+/+ mice. Taken together, these data demonstrate that IDH2 plays a protective role against UUO-induced inflammation through inhibition of oxidative stress and macrophage infiltration.

Differentially Expressed Genes in Period 2-Overexpressing Mice Striatum May Underlie Their Lower Sensitivity to Methamphetamine Addiction-Like Behavior

  • Sayson, Leandro Val;Kim, Mikyung;Jeon, Se Jin;Custodio, Raly James Perez;Lee, Hyun Jun;Ortiz, Darlene Mae;Cheong, Jae Hoon;Kim, Hee Jin
    • Biomolecules & Therapeutics
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    • v.30 no.3
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    • pp.238-245
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    • 2022
  • Previous reports have demonstrated that genetic mechanisms greatly mediate responses to drugs of abuse, including methamphetamine (METH). The circadian gene Period 2 (Per2) has been previously associated with differential responses towards METH in mice. While the behavioral consequences of eliminating Per2 have been illustrated previously, Per2 overexpression has not yet been comprehensively described; although, Per2-overexpressing (Per2 OE) mice previously showed reduced sensitivity towards METH-induced addiction-like behaviors. To further elucidate this distinct behavior of Per2 OE mice to METH, we identified possible candidate biomarkers by determining striatal differentially expressed genes (DEGs) in both drug-naïve and METH-treated Per2 OE mice relative to wild-type (WT), through RNA sequencing. Of the several DEGs in drug naïve Per2 OE mice, we identified six genes that were altered after repeated METH treatment in WT mice, but not in Per2 OE mice. These results, validated by quantitative real-time polymerase chain reaction, could suggest that the identified DEGs might underlie the previously reported weaker METH-induced responses of Per2 OE mice compared to WT. Gene network analysis also revealed that Asic3, Hba-a1, and Rnf17 are possibly associated with Per2 through physical interactions and predicted correlations, and might potentially participate in addiction. Inhibiting the functional protein of Asic3 prior to METH administration resulted in the partial reduction of METH-induced conditioned place preference in WT mice, supporting a possible involvement of Asic3 in METH-induced reward. Although encouraging further investigations, our findings suggest that these DEGs, including Asic3, may play significant roles in the lower sensitivity of Per2 OE mice to METH.

Characterization of age- and stage-dependent impaired adult subventricular neurogenesis in 5XFAD mouse model of Alzheimer's disease

  • Hyun Ha Park;Byeong-Hyeon Kim;Seol Hwa Leem;Yong Ho Park;Hyang-Sook Hoe;Yunkwon Nam;Sujin Kim;Soo Jung Shin;Minho Moon
    • BMB Reports
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    • v.56 no.9
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    • pp.520-525
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    • 2023
  • Alzheimer's disease (AD) is a progressive neurodegenerative disease characterized by cognitive decline. Several recent studies demonstrated that impaired adult neurogenesis could contribute to AD-related cognitive impairment. Adult subventricular zone (SVZ) neurogenesis, which occurs in the lateral ventricles, plays a crucial role in structural plasticity and neural circuit maintenance. Alterations in adult SVZ neurogenesis are early events in AD, and impaired adult neurogenesis is influenced by the accumulation of intracellular Aβ. Although Aβ-overexpressing transgenic 5XFAD mice are an AD animal model well representative of Aβ-related pathologies in the brain, the characterization of altered adult SVZ neurogenesis following AD progression in 5XFAD mice has not been thoroughly examined. Therefore, we validated the characterization of adult SVZ neurogenesis changes with AD progression in 2-, 4-, 8-, and 11-monthold male 5XFAD mice. We first investigated the Aβ accumulation in the SVZ using the 4G8 antibody. We observed intracellular Aβ accumulation in the SVZ of 2-month-old 5XFAD mice. In addition, 5XFAD mice exhibited significantly increased Aβ deposition in the SVZ with age. Next, we performed a histological analysis to investigate changes in various phases of adult neurogenesis, such as quiescence, proliferation, and differentiation, in SVZ. Compared to age-matched wild-type (WT) mice, quiescent neural stem cells were reduced in 5XFAD mice from 2-11 months of age. Moreover, proliferative neural stem cells were decreased in 5XFAD mice from 2 to 8 months of age. Furthermore, differentiations of neuroblasts were diminished in 5XFAD mice from 2-11 months of age. Intriguingly, we found that adult SVZ neurogenesis was reduced with aging in healthy mice. Taken together, our results revealed that impairment of adult SVZ neurogenesis appears with aging or AD progression.

Gene expression profiles of skin from cyclin dependent kinases 5-knockdown mice

  • Shanshan Yang;Dingxing Jiao;Tao Song;Ping Rui;Ruiwen Fan;Zengjun Ma
    • Animal Bioscience
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    • v.37 no.4
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    • pp.567-575
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    • 2024
  • Objective: This study aimed to identify genes regulated by cyclin dependent kinases 5 (CDK5) that participate in hair pigmentation in mice. Methods: The mRNA expression profiles of skin samples from CDK5-knockdown mice were constructed using high-throughput RNA sequencing and compared with those of wild-type mice. Results: In total, 8,002 known genes were differentially expressed between CDK5-knockdown and wild-type mice. Of these, 3,658 were upregulated and 4,344 were downregulated in the skin of CDK5-knockdown mice. An additional 318 previously unknown genes were also differentially expressed, with 171 downregulated and 147 upregulated genes in the skin of CDK5-knockdown mice. Of the known genes expressed in mouse skin, 80 were associated with hair color, with 61 showing lower expression and 19 exhibiting higher expression in skin of CDK5-knockdown mice. Importantly, the expression of the tyrosinase-related protein 1 (TYRP1) and the calcium signaling pathway were also found to be regulated by CDK5, suggesting that pigmentation is regulated by CDK5 via the calcium signaling pathway and TYRP1. Conclusion: The transcriptome profiles obtained from the skin of CDK5-knockdown mice compared to wild-type mice provide a valuable resource to help understand the mechanism by which CDK5 regulates melanogenesis in mice and other animals.

Pharmacological Studies on Melilotus officinalis Extract (Melilotus officinalis 엑기스의 약리학적(藥理學的) 연구(硏究))

  • Hong, N.D.;Won, D.H.;Kim, N.J.
    • Korean Journal of Pharmacognosy
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    • v.14 no.2
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    • pp.51-59
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    • 1983
  • Experimental studies were made with Melilotus officinalis extract which was extracted from flowers and leaves of Melilotus officinalis Dsr. (Leguminosae). In this paper, acute toxicity, analgesic action, prolongation of hypnosis time by induced pentobarbital-Nain mice, antiinflammatory effect in rats and effects on isolated intestines of mice and rats were studied, The result was as follows; 1. Very low toxicity in mice. 2. Analgesic action was recognized markedly in mice. 3. Prolongation of hypnosis time induced by pentobarbital-Na in mice was shown. 4. Relaxing action was shown on the isolated ileum in mice and antagonistic action was seen on $BaCl_2-induced$ contraction of the ileum that the relaxing effect of the intestinal smooth muscle was recognized. 5.Antiinflammatory effect was shown markedly in mice. 6.Hypotensive and vaso-dilating actions due to the vascular smooth muscle relaxation were noted in rabbits.

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