• Title/Summary/Keyword: in-vitro physical model

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Melittin induces autophagy to alleviate chronic renal failure in 5/6-nephrectomized rats and angiotensin II-induced damage in podocytes

  • Yufan Zhang;Huaping Xu;Hongwei Qiao;Ya Zhao;Minmin Jiang
    • Nutrition Research and Practice
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    • v.18 no.2
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    • pp.210-222
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    • 2024
  • BACKGROUND/OBJECTIVES: Chronic renal failure (CRF) is a complex pathological condition that lacks a cure. Certain Chinese medicines, such as melittin, a major component in bee venom, have shown efficacy in treating CRF patients. On the other hand, the mechanisms underlying the therapeutic effects of melittin are unclear. MATERIALS/METHODS: A 5/6 nephrectomy model (5/6 Nx) of renal failure was established on rats for in vivo assays, and mouse podocyte clone 5 (MPC5) mouse podocyte cells were treated with angiotensin II (AngII) to establish an in vitro podocyte damage model. The 24-h urine protein, serum creatinine, and blood urea nitrogen levels were evaluated after one, 2, and 4 weeks. Hematoxylin and eosin staining, Masson staining, and periodic acid-Schiff staining were used to examine the pathological changes in kidney tissues. A cell counting kit 8 assay was used to assess the cell viability. Reverse transcription polymerase chain reaction and Western blot were used to assess the mRNA and protein levels in the cells, respectively. RESULTS: In the rat 5/6 Nx, melittin reduced the 24-h urinary protein excretion and the serum creatinine and blood urea nitrogen levels. Furthermore, the renal pathology was improved in the melittin-treated 5/6 Nx rats. Melittin promoted podocin, nephrin, Beclin 1, and the LC3II/LC3I ratio and inhibited phosphorylated mammalian target of rapamycin (mTOR)/mTOR in 5/6 Nx-induced rats and AngII-induced MPC5 mouse podocyte cells. Moreover, inhibiting autophagy with 3-MA weakened the effects of melittin on podocin, nephrin, and the LC3II/LC3I ratio in podocytes. CONCLUSION: Melittin may offer protection against kidney injury, probably by regulating podocyte autophagy. These results provide the theoretical basis for applying melittin in CRF therapy.

Lomens-P0 (mixed extracts of Hordeum vulgare and Chrysanthemum zawadskii) regulate the expression of factors affecting premenstrual syndrome symptoms

  • Lee, Yoon Seo;Jeon, Hyelin;Her, Yang-Mi;Lee, Da Eun;Jeong, Yong Joon;Kim, Eun Jeong;Choe, Tae Hwan;Suh, Hee Ju;Shin, Seung-Yeon;Park, Dae Won;Lee, Yeong-Geun;Kang, Se Chan
    • Nutrition Research and Practice
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    • v.15 no.6
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    • pp.715-731
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    • 2021
  • BACKGROUND/OBJECTIVES: Premenstrual syndrome (PMS) is a disorder characterized by repeated emotional, behavioral, and physical symptoms before menstruation, and the exact cause and mechanism are uncertain. Hyperprolactinemia interferes with the normal production of estrogen and progesterone, leading to PMS symptoms. Thus, we judged that the inhibition of prolactin hypersecretion could mitigate PMS symptoms. MATERIALS/METHODS: Hordeum vulgare L. extract (HVE), Chrysanthemum zawadskii var. latilobum extract (CZE), and Lomens-P0 the mixture of these extracts were tested in subsequent experiments. The effect of extracts on prolactin secretion at the in vitro level was measured in GH3 cells. Nitric oxide and pro-inflammatory mediator expression were measured in RAW 264.7 cells to confirm the anti-inflammatory effect. Also, the hyperprolactinemic Institute for Cancer Research (ICR) mice model was used to measure extract effects on prolactin and hormone secretion and uterine inflammation. RESULTS: Anti-inflammatory effects of and prolactin secretion suppress by HVE and CZE were confirmed through in vitro experiments (P < 0.05). Treatment with Lomens-P0 inhibited prolactin secretion (P < 0.05) and restored normal sex hormone secretion in the hyperprolactinemia mice model. In addition, extracts significantly inhibited the expression of pro-inflammatory biomarkers, including interleukin-1𝛽, and -6, tumor necrosis factor-𝛼, inducible nitric oxide synthase, and cyclooxygenase-2 (P < 0.01). We used high-performance liquid chromatography analyses to identify tricin and chlorogenic acid as the respective components of HVE and CZE that inhibit prolactin secretion. The Lomens-P0, which includes tricin and chlorogenic acid, is expected to be effective in improving PMS symptoms in the human body. CONCLUSIONS: The Lomens-P0 suppressed the prolactin secretion in hyperprolactinemia mice, normalized the sex hormone imbalance, and significantly suppressed the expression of inflammatory markers in uterine tissue. This study suggests that Lomens-P0 may have the potential to prevent or remedy materials to PMS symptoms.

Surface Observation of Mg-HA Coated Ti-6Al-4V Alloy by Plasma Electrolytic Oxidation

  • Yu, Ji-Min;Choe, Han-Cheol
    • Proceedings of the Korean Institute of Surface Engineering Conference
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    • 2016.11a
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    • pp.198-198
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    • 2016
  • An ideal orthopedic implant should provide an excellent bone-implant connection, less implant loosening and minimum adverse reactions. Commercial pure titanium (CP-Ti) and Ti alloys have been widely utilized for biomedical applications such as orthopedic and dental implants. However, being bioinert, the integration of such implant in bone was not in good condition to achieve improved osseointegraiton, there have been many efforts to modify the composition and topography of implant surface. These processes are generally classified as physical, chemical, and electrochemical methods. Plasma electrolytic oxidation (PEO) as an electrochemical route has been recently utilized to produce this kind of composite coatings. Mg ion plays a key role in bone metabolism, since it influences osteoblast and osteoclast activity. From previous studies, it has been found that Mg ions improve the bone formation on Ti alloys. PEO is a promising technology to produce porous and firmly adherent inorganic Mg containing $TiO_2$($Mg-TiO_2$ ) coatings on Ti surface, and the amount of Mg introduced into the coatings can be optimized by altering the electrolyte composition. In this study, a series of $Mg-TiO_2$ coatings are produced on Ti-6Al-4V ELI dental implant using PEO, with the substitution degree, respectively, at 0, 5, 10 and 20%. Based on the preliminary analysis of the coating structure, composition and morphology, a bone like apatite formation model is used to evaluate the in vitro biological responses at the bone-implant interface. The enhancement of the bone like apatite forming ability arises from $Mg-TiO_2$ surface, which has formed the reduction of the Mg ions. The promising results successfully demonstrate the immense potential of $Mg-TiO_2$ coatings in dental and biomaterials applications.

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Validation and comparison of volume measurements using 1 multidetector computed tomography and 5 cone-beam computed tomography protocols: An in vitro study

  • Juliana Andrea Correa, Travessas;Alessandra Mendonca, dos Santos;Rodrigo Pagliarini, Buligon;Nadia Assein, Arus;Priscila Fernanda Tiecher, da Silveira;Heraldo Luis Dias, da Silveira;Mariana Boessio, Vizzotto
    • Imaging Science in Dentistry
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    • v.52 no.4
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    • pp.399-408
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    • 2022
  • Purpose: The purpose of this study was to compare volume measurements obtained using 2 image software packages on Digital Imaging and Communications in Medicine (DICOM) images acquired from 1 multidetector computed tomography and 5 cone-beam computed tomography devices, using different protocols for physical volume measurements. Materials and Methods: Four pieces of bovine leg were prepared. Marrow was removed from 3 pieces, leaving cortical bone exposed. The resulting space of 1 piece was filled with water, another was filled with propylene glycol, and the third was left unfilled. The marrow in the fourth sample was left fully intact. Volume measurements were obtained after importing DICOM images into the Dolphin Imaging 11.95 and ITK-SNAP software programs. Data were analyzed using 3-way analysis of variance with a generalized linear model to determine the effects of voxel size, software, and content on percentage mean volume differences between tomographic protocols. A significance level of 0.05 was used. Results: The intraclass correlation coefficients for intraobserver and interobserver reliability were, respectively, 0.915 and 0.764 for the Dolphin software and 0.894 and 0.766 for the ITK-SNAP software. Three sources of statistically significant variation were identified: the interaction between software and content (P=0.001), the main effect of content (P=0.014), and the main effect of software (P=0.001). Voxel size was not associated with statistically significant differences in volume measurements. Conclusion: Both content and software influenced the accuracy of volume measurements, especially when the content had gray values similar to those of the adjacent tissues.

Continuous DC-CIK Infusions Restore CD8+ Cellular Immunity, Physical Activity and Improve Clinical Efficacy in Advanced Cancer Patients Unresponsive to Conventional Treatments

  • Zhao, Yan-Jie;Jiang, Ni;Song, Qing-Kun;Wu, Jiang-Ping;Song, Yu-Guang;Zhang, Hong-Mei;Chen, Feng;Zhou, Lei;Wang, Xiao-Li;Zhou, Xin-Na;Yang, Hua-Bing;Ren, Jun;Lyerly, Herbert Kim
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.6
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    • pp.2419-2423
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    • 2015
  • Background: There are few choices for treatment of advanced cancer patients who do not respond to or tolerate conventional anti-cancer treatments. Therefore this study aimed to deploy the benefits and clinical efficacy of continuous dendritic cell-cytokine induced killer cell infusions in such patients. Materials and Methods: A total of 381 infusions (from 67 advanced cases recruited) were included in this study. All patients underwent peripheral blood mononuclear cell apheresis for the following cellular therapy and dendritic cells-cytokine induced killer cells were expanded in vitro. Peripheral blood T lymphocyte subsets were quantified through flow cytometry to address the cellular immunity status. Clinical efficacy and physical activities were evaluated by RECIST criteria and Eastern Cooperative Oncology Group scores respectively. Logistic regression model was used to estimate the association between cellular infusions and clinical benefits. Results: An average of $5.7{\pm}2.94{\times}10^9$ induced cells were infused each time and patients were exposed to 6 infusions. Cellular immunity was improved in that cytotoxic $CD8^+CD28^+$ T lymphocytes were increased by 74% and suppressive $CD8^+CD28^-$ T lymphocytes were elevated by 16% (p<0.05). Continuous infusion of dendritic cells-cytokine induced killer cells was associated with improvement of both patient status and cellular immunity. A median of six infusions were capable of reducing risk of progression by 70% (95%CI 0.10-0.91). Every elevation of one ECOG score corresponded to a 3.90-fold higher progression risk (p<0.05) and 1% increase of $CD8^+CD28^-$ T cell proportion reflecting a 5% higher risk of progression (p<0.05). Conclusions: In advanced cancer patients, continuous dendritic cell-cytokine induced killer cell infusions are capable of recovering cellular immunity, improving patient status and quality of life in those who are unresponsive to conventional cancer treatment.

Ginsenoside Rg3-enriched red ginseng extract inhibits platelet activation and in vivo thrombus formation

  • Jeong, Dahye;Irfan, Muhammad;Kim, Sung-Dae;Kim, Suk;Oh, Jun-Hwan;Park, Chae-Kyu;Kim, Hyun-Kyoung;Rhee, Man Hee
    • Journal of Ginseng Research
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    • v.41 no.4
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    • pp.548-555
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    • 2017
  • Background: Korean Red Ginseng has been used for several decades to treat many diseases, enhancing both immunity and physical strength. Previous studies have documented the therapeutic effects of ginseng, including its anticancer, antiaging, and anti-inflammatory activities. These activities are mediated by ginsenosides present in the ginseng plant. Ginsenoside Rg3, an effective compound from red ginseng, has been shown to have antiplatelet activity in addition to its anticancer and anti-inflammatory activities. Platelets are important for both primary hemostasis and the repair of the vessels after injury; however, they also play a crucial role in the development of acute coronary diseases. We prepared ginsenoside Rg3-enriched red ginseng extract (Rg3-RGE) to examine its role in platelet physiology. Methods: To examine the effect of Rg3-RGE on platelet activation in vitro, platelet aggregation, granule secretion, intracellular calcium ($[Ca^{2+}]_i$) mobilization, flow cytometry, and immunoblot analysis were carried out using rat platelets. To examine the effect of Rg3-RGE on platelet activation in vivo, a collagen plus epinephrine-induced acute pulmonary thromboembolism mouse model was used. Results: We found that Rg3-RGE significantly inhibited collagen-induced platelet aggregation and $[Ca^{2+}]_i$ mobilization in a dose-dependent manner in addition to reducing ATP release from collagen-stimulated platelets. Furthermore, using immunoblot analysis, we found that Rg3-RGE markedly suppressed mitogen-activated protein kinase phosphorylation (i.e., extracellular stimuli-responsive kinase, Jun N-terminal kinase, p38) as well as the PI3K (phosphatidylinositol 3 kinase)/Akt pathway. Moreover, Rg3-RGE effectively reduced collagen plus epinephrine-induced mortality in mice. Conclusion: These data suggest that ginsenoside Rg3-RGE could be potentially be used as an antiplatelet therapeutic agent against platelet-mediated cardiovascular disorders.

THE EFFECTS OF BIO-$OSS^{(R)}$ AS A SCAFFOLDS DURING SINUS BONE GRAFT USING MESENCHYMAL STEM CELLS IN RABBIT (가토에서 자가유래 골아줄기세포를 이용한 상악동 골 이식술시 비계체로서 Bio-$Oss^{(R)}$의 효과에 관한 연구)

  • Lee, Jun;Sung, Dae-Hyuk;Choi, Jae-Young;Choi, Sung-Rym;Cha, Su-Ryun;Jang, Jae-Deog;Kim, Eun-Chul
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.33 no.5
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    • pp.405-418
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    • 2007
  • Mesenchymal stem cells(MSCs) have been though to be multipotent cells that can replicate that have the potential to differentiate into lineages of mesenchymal tissue including the bone, cartilage, fat, tendon, muscle, and marrow stroma. Especially, scaffolds to support cell-based tissue engineering are critical determinants of clinical efforts to regenerate and repair the body. Selection of a matrix carrier imvolves consideration of the matrix's role as a scaffold for physical support and host tissue integration as well as its ability to support of synergize the osteoinductive program of the implanted mesenchymal stem cell. The aim of this study is to evaluate the effect of autobone and Bio-$Oss^{(R)}$ to adherent mesenchymal stem cells as scaffolds on sinus augmentation with fibrin glue mixture in a rabbit model. 16 New Zealand White rabbits were divided randomly into 4 groups based on their time of sacrifice(1, 2, 4 and 8 weeks). First, mesenchymal stem cells were isolated from iliac crest marrow of rabbits and expanded in vitro. Cell culture was performed in accordance with the technique described by Tsutsumi et al. In the present study, the animals were sacrificed at 1, 2, 4 and 8 weeks after transplantation, and the bone formation ability of each sides was evaluated clinically, radiologically, histologically and histomorphologically. According to the histological observations, autobone scaffolds group showed integrated graft bone with host bone from sinus wall. At 2 and 4 weeks, it showed active newly formed bone and neovascularization. At 8 weeks, lamellae bone was observed in sinus graft material area. Radiologically, autobone with stem cell showed more radiopaque than Bio-$Oss^{(R)}$ scaffolds group. there were significant differences in bone volume between 4 and 8 weeks(p<0.05).