• Title/Summary/Keyword: Blood Assay

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Combination stem cell therapy using dental pulp stem cells and human umbilical vein endothelial cells for critical hindlimb ischemia

  • Kim, Chung Kwon;Hwang, Ji-Yoon;Hong, Tae Hee;Lee, Du Man;Lee, Kyunghoon;Nam, Hyun;Joo, Kyeung Min
    • BMB Reports
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    • v.55 no.7
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    • pp.336-341
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    • 2022
  • Narrowing of arteries supplying blood to the limbs provokes critical hindlimb ischemia (CLI). Although CLI results in irreversible sequelae, such as amputation, few therapeutic options induce the formation of new functional blood vessels. Based on the proangiogenic potentials of stem cells, in this study, it was examined whether a combination of dental pulp stem cells (DPSCs) and human umbilical vein endothelial cells (HUVECs) could result in enhanced therapeutic effects of stem cells for CLI compared with those of DPSCs or HUVECs alone. The DPSCs+ HUVECs combination therapy resulted in significantly higher blood flow and lower ischemia damage than DPSCs or HUVECs alone. The improved therapeutic effects in the DPSCs+ HUVECs group were accompanied by a significantly higher number of microvessels in the ischemic tissue than in the other groups. In vitro proliferation and tube formation assay showed that VEGF in the conditioned media of DPSCs induced proliferation and vessel-like tube formation of HUVECs. Altogether, our results demonstrated that the combination of DPSCs and HUVECs had significantly better therapeutic effects on CLI via VEGF-mediated crosstalk. This combinational strategy could be used to develop novel clinical protocols for CLI proangiogenic regenerative treatments.

Investigating the Potential of Lipids for Use as Biomarkers for Glioblastoma via an Untargeted Lipidomics Approach

  • Burcak Soylemez;Zekeriya Bulut;Serap Sahin-Bolukbasi
    • Journal of Korean Neurosurgical Society
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    • v.66 no.2
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    • pp.133-143
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    • 2023
  • Objective : The types and functions of lipids involved in glioblastoma (GB) are not well known. Lipidomics is a new field that examines cellular lipids on a large scale and novel aplication of lipidomics in the biomedical sciences have emerged. This study aimed to investigate the potential of blood lipids for use as biomarkers for the diagnosis of GB via untargated lipidomic approach. Gaining a deeper understanding of lipid metabolism in patients with GB can contribute to the early diagnosis with GB patiens and also development of novel and better therapeutic options. Methods : This study was performed using blood samples collected from 14 patients (eight females and six males) and 14 controls (eight females and six males). Lipids were extracted from blood samples and quantified using phosphorus assay. Lipid profiles of between patients with GB and controls were compared via an untargeted lipidomics approach using 6530 Accurate-Mass Q-TOF LC/MS mass spectrometer. Results : According to the results obtained using the untargeted lipidomics approach, differentially regulated lipid species, including fatty acid (FA), glycerolipid (GL), glycerophospholipid (PG), saccharolipid (SL), sphingolipid (SP), and sterol lipid (ST) were identified between in patients with GB and controls. Conclusion : Differentially regulated lipids were identified in patients with GB, and these lipid species were predicted as potential biomarkers for diagnosis of GB.

Host Blood Transcriptional Signatures as Candidate Biomarkers for Predicting Progression to Active Tuberculosis

  • Chang Ho Kim;Gahye Choi;Jaehee Lee
    • Tuberculosis and Respiratory Diseases
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    • v.86 no.2
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    • pp.94-101
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    • 2023
  • A recent understanding of the dynamic continuous spectrum of Mycobacterium tuberculosis infection has led to the recognition of incipient tuberculosis, which refers to the latent infection state that has begun to progress to active tuberculosis. The importance of early detection of these individuals with a high-risk of progression to active tuberculosis is emphasized to efficiently implement targeted tuberculosis preventive therapy. However, the tuberculin skin test or interferon-γ release assay, which is currently used for the diagnosis of latent tuberculosis infection, does not aid in the prediction of the risk of progression to active tuberculosis. Thus, a novel test is urgently needed. Recently, simultaneous and systematic analysis of differentially expressed genes using a high-throughput platform has enabled the discovery of key genes that may serve potential biomarkers for the diagnosis or prognosis of diseases. This host transcriptional investigation has been extended to the field of tuberculosis, providing promising results. The present review focuses on recent progress and challenges in the field of blood transcriptional signatures to predict progression to active tuberculosis.

Isolation of Peripheral Blood-Derived Mesenchymal Stem Cells in Mares and Foals

  • Ye-Eun Oh;Eun-Bee Lee;Jong-Pil Seo
    • Journal of Veterinary Clinics
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    • v.40 no.5
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    • pp.323-329
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    • 2023
  • Peripheral blood-derived mesenchymal stem cells (PB-MSCs) have shown promise in cell-based therapy, as they can be harvested with ease through minimally invasive procedures. This study aimed to isolate PB-MSCs from foals and mares and to compare the proliferation and cellular characteristics of the PB-MSCs between the two groups. Six pairs of mares and their foals were used in this study. MSCs were isolated from PB by direct plating in a tissue culture medium, and cell proliferation (population doubling time [PDT], and colony-forming unit-fibroblast assay [CFU-F]), and characterization (morphology, plastic adhesiveness, colony formation, trilineage differentiation) were examined. There was no significant difference in the PB-MSC yield, CFU-F, and PDT between the mares and foals. PB-MSCs from both mares and foals showed typical MSC characteristics in terms of spindle-shaped morphology, plastic adhesive properties, formation of colonies, trilineage differentiation. These results suggest that PB-MSCs isolated from horses, both adult horses, and foals, can be used for equine cell-based therapy.

Drug Interaction of Cimetidine and Cyclosporine in Human (정상지원자에서 Cimetidine과 Cyclosporine의 약물상호작용)

  • Choi, In;Choi, Jun Shik
    • Korean Journal of Clinical Pharmacy
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    • v.7 no.2
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    • pp.51-63
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    • 1997
  • The effect of cimetidine administration on the pharmacokinetic parameters of cyclosporine were determined in healthy voluteers. This study was performed in 10 volunteers of age ranged 22-48 years and body weight 48-62 kg. This study was performed with cross-over design. Mono cyclosporine and cyclosporine metabolites was extracted from whole blood analysed by fluororescence polarization immune assay (TDX-FLX, Abbott). After coadministration of cimetidine (300 mg) with cyclosporine (300 mg) orally, maximum concentration of mono cyclosporine was significantly increased $1221{\pm}143\;ng/ml\;to\;1562{\pm}184\;ng/ml$ (P<0.05), area under the time curve of cyclosporine (12 hr) also was significantly increased $7478{\pm}829\;ng/ml{\cdot}hr\;to\;9721{\pm}879\;ng/ml{\cdot}hr$ (P<0.05) and absolute baioavailability of cyclosporine was increased $50\pm5.6\%\;to\;57.6\pm6.1\%\;(P<0.05)$ compared to control group. The blood concentrations of cyclopsorine metabolites were significantly decrased (P<0.05) after coadministration of cimetidine. In cimetidine pretreated group, blood mono cyclosporine concentrations were increased significan시y $1220.0\pm203.00\;ng/ml\;to\;1510.0\pm204.00\;ng/ml$ compared with control group (P<0.05). In the mono cyclosporine pharmacokinetic parameter after oral administration absorption rate and maximum concentration were significantly higher in cimetidine coadministered and pretreated group than control group (P<0.05). The ratio of metabolites and mono cyclosporine concentrations was decreased significantly from $70.8\%\;in\;control\;to\;34.8\%$ in coadministration of cimetidine orally. As matter of facts these reults are considered to inhibition of cyclosporine hepatic metabolism and increasing of cyclosporine absorption rate in gastrointestinal tract because of maintaining cyclosporine stability in elevated gastric pH by cimetidine. We considered, it appeares that cimetidine increase bioavailability of cyclosporine by increasing oral absorption and by decreasing hepatic clearance. But the absorption and clearance of cyclosporine was highly variable individually, and therefore we consider that cyclosporine blood level monitoring would be essential in patients with cimetidine co-administration.

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Physalin D inhibits RANKL-induced osteoclastogenesis and bone loss via regulating calcium signaling

  • Ding, Ning;Lu, Yanzhu;Cui, Hanmin;Ma, Qinyu;Qiu, Dongxia;Wei, Xueting;Dou, Ce;Cao, Ning
    • BMB Reports
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    • v.53 no.3
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    • pp.154-159
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    • 2020
  • We investigated the effects of physalin A, B, D, and F on osteoclastogenesis induced by receptor activator of nuclear factor κB ligand (RANKL). The biological functions of different physalins were first predicted using an in silico bioinformatic tool (BATMAN-TCM). Afterwards, we tested cell viability and cell apoptosis rate to analyze the cytotoxicity of different physalins. We analyzed the inhibitory effects of physalins on RANKL-induced osteoclastogenesis from mouse bone-marrow macrophages (BMMs) using a tartrate-resistant acid phosphatase (TRAP) stain. We found that physalin D has the best selectivity index (SI) among all analyzed physalins. We then confirmed the inhibitory effects of physalin D on osteoclast maturation and function by immunostaining of F-actin and a pit-formation assay. On the molecular level, physalin D attenuated RANKL-evoked intracellular calcium ([Ca(2+)](i)) oscillation by inhibiting phosphorylation of phospholipase Cγ2 (PLCγ2) and thus blocked the downstream activation of Ca2+/calmodulin-dependent protein kinases (CaMK)IV and cAMP-responsive element-binding protein (CREB). An animal study showed that physalin D treatment rescues bone microarchitecture, prevents bone loss, and restores bone strength in a model of rapid bone loss induced by soluble RANKL. Taken together, these results suggest that physalin D inhibits RANKL-induced osteoclastogenesis and bone loss via suppressing the PLCγ2-CaMK-CREB pathway.

Clinical Experience in Treatment of Diabetic Foot Ulcers Using Platelet Concentrates from Blood Bank (혈액은행 제조의 혈소판 농축액을 이용한 당뇨족부궤양의 치료 경험)

  • Kim, Deok Woo;Han, Seung Kyu;Kim, Woo Kyung
    • Archives of Plastic Surgery
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    • v.33 no.2
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    • pp.198-204
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    • 2006
  • Many clinical trials have shown the effectiveness of the platelet releasate or the platelet gel on chronic wounds. However, the patient's own blood had to be aspirated and processed to make the platelet releasate or a platelet gel. The purpose of this study was to assess the effects of platelet concentrates from the blood bank for the treatment of diabetic foot ulcers. To obtain the basic data of the PDGF-BB content in platelet concentrates supplied from the blood bank, enzyme-linked immunosorbent assay quantification was performed. On average, 8.5 pg of the PDGF-BB was released per 1 million platelets. Sixteen patients with diabetic foot ulcers ranging from 1.0 to $18.0cm^2$(mean, $6.1cm^2$) in size were treated. The platelet concentrates was centrifuged and the precipitantte was mixed with 1 ml of fibrinogen. The platelets and fibrinogen mixture was dispersed on to the ulcer lesions. The liquid platelet and fibrinogen mixture was then sealed using 0.3-1.0 ml of thrombin and moisture dressing was performed. The procedure was repeated every one or two weeks until wound closure. Time required for complete healing ranged from 3 to 12 weeks after treatment (mean, 7.3 weeks). Patient satisfaction was also very positive. In this study, the use of platelet concentrates from the blood bank was found to be effective in treating diabetic foot ulcers.

Exercise induced upregulation of glutamate-cysteine ligase catalytic subunit and glutamate-cysteine ligase modifier subunit gene expression in Thoroughbred horses

  • Park, Jeong-Woong;Choi, Jae-Young;Hong, Seul A;Kim, Nam Young;Do, Kyoung-Tag;Song, Ki-Duk;Cho, Byung-Wook
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.5
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    • pp.728-735
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    • 2017
  • Objective: This study was performed to reveal the molecular structure and expression patterns of horse glutamate-cysteine ligase catalytic subunit (GCLC) and glutamate-cysteine ligase modifier subunit (GCLM) genes whose products form glutamate cysteine ligase, which were identified as differentially expressed genes in the previous study. Methods: We performed bioinformatics analyses, and gene expression assay with quantitative polymerase chain reaction (qPCR) for horse GCLC and GCLM genes in muscle and blood leukocytes of Thoroughbred horses Results: Expression of GCLC showed the same pattern in both blood and muscle tissues after exercise. Expression of GCLC increased in the muscle and blood of Thoroughbreds, suggesting a tissue-specific regulatory mechanism for the expression of GCLC. In addition, expression of the GCLM gene increased after exercise in both the blood and muscle of Thoroughbreds. Conclusion: We established the expression patterns of GCLC and GCLM in the skeletal muscle and blood of Thoroughbred horses in response to exercise. Further study is now warranted to uncover the functional importance of these genes in exercise and recovery in racehorses.

The Antidiabetic Effects of Black Ginseng Extract(BGE)and Geumsan Black Ginseng 05-FF(GBG05-FF) on In Vitro and In Vivo Assay (흑삼추출물(BGE)과 금산흑삼표준화소재(GBG05-FF)의 In Vitro와 In Vivo상에서의 항당뇨효과)

  • Seo, Yun-Soo;Shon, Mi-Yae;Kong, Ryong;Kang, Ok-Hwa;Zhou, Tian;Kim, Do-Yeon;Park, Jong-Dae;Kwon, Dong-Yeul
    • Korean Journal of Pharmacognosy
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    • v.46 no.4
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    • pp.321-326
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    • 2015
  • The prevalence of diabetes mellitus continues to rise alarmingly because of industrialization of society. The aim of this study was to investigate the antidiabetic effects of black ginseng extract (BGE) which was performed panax ginseng. First we examined the inhibitory effects of BGE on ${\alpha}$-glucosidase. But there was no practical effects in our observations. We, also, investigate the effects of BGE on glucose uptake of skeletal muscle using 2-NBDG in $C_2C_{12}$ myotube. BGE significantly improved the glucose uptake considered as a lowered blood glucose level. Effects of GBG05-FF on fasting blood glucose and glycated hemoglobin (HbA1c) were investigated in streptozotocin (STZ)-induced diabetic mouse. After injection of STZ, fasting blood glucose and glycated hemoglobin rapidly increased. But STZ-induced diabetic mouse treated with GBG05-FF significantly reduced the level of fasting blood glucose and HbA1c. This results showed that supplementation of BGE improve the diabetic parameters and BGE have a potentiality as a functional food for Diabetes mellitus.

Zinc Enhances Neutrophil Extracellular Trap Formation of Porcine Peripheral Blood Polymorphonuclear Cells through Tumor Necrosis Factor-Alpha from Peripheral Blood Mononuclear Cells

  • Heo, Ju-Haeng;Kim, Hakhyun;Kang, Byeong-Teck;Yang, Mhan-Pyo
    • Journal of Veterinary Clinics
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    • v.37 no.5
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    • pp.249-254
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    • 2020
  • Neutrophil extracellular trap (NET) formation is an immune response for the invasion of microbes. The purpose of this study is to examine the effect of zinc on NET formation of porcine peripheral blood polymorphonuclear cells (PMNs). The NET formation of PMNs was measured by fluorescence microplate reader. The production of tumor necrosis factor (TNF)-α in the culture supernatants from zinc-treated peripheral blood mononuclear cells (PBMCs) was measured by enzyme-linked immunosorbent assay (ELISA). Zinc itself did not have no effect on NET formation. However, the NET formation of PMNs was increased by culture supernatants from PBMCs treated with zinc. Also, the NET formation of PMNs was increased by recombinant porcine (rp) TNF-α. The production of TNF-α in PBMCs culture supernatants was shown to increase upon zinc treatments. These NET formations of PMNs increased by either culture supernatant from PBMCs treated with zinc or rpTNF-α were inhibited by treatment of anti-rpTNF-α polyclonal antibody (pAb). These results suggested that zinc has an immunostimulating effect on the NET formation of PMNs, which is mediated by TNF-α released from zinc-treated PBMCs. Therefore, zinc may play an important role for NET formation in the defense of porcine inflammatory diseases.