• Title/Summary/Keyword: Molecular diagnostics

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Fused Polypeptide with DEF Induces Apoptosis of Lung Adenocarcinoma Cells

  • Liang, Ai-Ling;Zhang, Ting-Ting;Zhou, Ning;Huang, Di-Nan;Liu, Xin-Guang;Liu, Yong-Jun;Tu, Zhi-Guang
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7339-7344
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    • 2013
  • To analyze the effects of a new unknown peptide DEF on the growth of tumor cells, a fused polypeptide TAT-DV1-DEF was designed and synthesized. The lung adenocarcinoma cell line GLC-82 treated with TAT-DV1-DEF was analyzed with a cell counting kit 8, and the location of polypeptides in cells was observed under laser confocal microscopy. The efficiency of polypeptide transfection and changes in nuclear morphology were analyzed by flow cytometry and fluorescence microscopy, respectively. Finally, the mechanism of tumor cell growth inhibition was evaluated by Western blotting. We found that TAT-DV1-DEF could significantly inhibit the growth of the lung adenocarcinoma cell line GLC-82, but not the normal human embryonic kidney cell line HEK-293. Polypeptides were found to be mostly localized in the cytoplasm and some mitochondria. The efficiency of polypeptide transfection in the two cell types was approximately 99%. Apoptotic nuclei were observed under fluorescence microscopy upon treatment with polypeptides and DAPI staining. Western blot analyses indicated that the polypeptide inhibition of tumor cell growth was apoptosis dependent. In the present study, we demonstrated that fused polypeptides could induce apoptosis of the lung adenocarcinoma cell line GLC-82, indicating that the new unknown peptide DEF has antitumor effects.

Slipchip Device Development in Molecular Diagnostics

  • Qingtian Yin;Huiwen Bai;Ruijie Li;Youngung Seok
    • Korean Journal of Materials Research
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    • v.34 no.2
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    • pp.63-71
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    • 2024
  • Slipchip offers advantages such as high-throughout, low cost, and simple operation, and therefore, it is one of the technologies with the greatest potential for high-throughput, single-cell, and single-molecule analyses. Slipchip devices have achieved remarkable advances over the past decades, with its simplified molecular diagnostics gaining particular attention, especially during the COVID-19 pandemic and in various infectious diseases scenarios. Medical testing based on nucleic acid amplification in the Slipchip has become a promising alternative simple and rapid diagnostic tool in field situations. Herein, we present a comprehensive review of Slipchip device advances in molecular diagnostics, highlighting its use in digital recombinase polymerase amplification (RPA), loop-mediated isothermal amplification (LAMP), and polymerase chain reaction (PCR). Slipchip technology allows users to conduct reliable droplet transfers with high-throughput potential for single-cell and molecule analyses. This review explores the device's versatility in miniaturized and rapid molecular diagnostics. A complete Slipchip device can be operated without special equipment or skilled handling, and provides high-throughput results in minimum settings. This review focuses on recent developments and Slipchip device challenges that need to be addressed for further advancements in microfluidics technology.

Immunostimulating Effects of Polysaccharides of Panax ginseng by Oral Administration into BALB/c Mice

  • Kang, Mi-Hyeon;Park, A-Young;Yang, Ae-Hee;Park, Ye-Eun;Hong, Seung-Hye;Park, Gab-Soon;Jung, Suk-Yul
    • Biomedical Science Letters
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    • v.20 no.4
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    • pp.194-200
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    • 2014
  • Ginsan, polysaccharide isolated from the root of Panax ginseng C.A. Meyer, has been shown to be a potent immunomodulator, producing several cytokines and stimulating lymphoid cells to proliferate. In this study, ginsan was orally inoculated into BALB/c mice up to 39 days and the activity of immune cells containing macrophages and T cells was analyzed. Moreover, the production of cytokines, e.g., tumor necrotic factor-${\alpha}$ (TNF-${\alpha}$), interferon-${\gamma}$ (IFN-${\gamma}$), GM-CSF and IL-12 was also analyzed. In results, the phagocytosis of macrophages was increased. About 13% cytotoxicity of NK cells was observed in 22 days and 29 days of administration. But, oral administration did not highly affect the proliferation of T cells. In cytokine analysis, 150 mg/kg and 300 mg/kg at 22 days and 29 days showed three times more increase in TNF-${\alpha}$ than the controls. IFN-${\gamma}$ showed 1.07 and 1.16 times more increase at 150 mg/kg and 300 mg/kg over 22 days, respectively more than the controls. 32 days of 150 mg/kg and 300 mg/kg induced GM-CSF of about 1.3 times more than the controls. IL-12 was not induced in samples more than the controls. Ginsan could be a potential immunostimulator. Therefore, our study suggests that it can be adapted as an immunostimulator that requires a relatively short oral administration.

Application of Molecular Diagnostics Technology in the Development of a Companion Diagnostics for Malignant Solid Tumors (악성 고형암의 항암제 동반진단 기술에서 분자진단기술의 적용)

  • Kim, Jin-Hee
    • The Journal of the Korea Contents Association
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    • v.19 no.3
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    • pp.365-374
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    • 2019
  • Unlike benign tumors, malignant tumors are capable of metastasis, easy to relapse, poor survival, and low quality of life. In Korea, here is a tendency to treat the tumors collectively according to the General Principles of Cancer Chemotherapy(GPCC) of the Health Insurance Review & Assessment Service (HIRA). But recently, companion diagnostics(CDx) is recommended rather than unilateral medication because biomarker-based molecular diagnostics is possible to predict the drug response of patients before drug treatment. Not only domestic but also overseas Food and Drug Administratio (FDA) recommends the development of the CDx system at the stage of drug development to ensure the responsiveness and safety of medicines. In this study, I focused on the necessity of CDx development direction as well as CDx development status through literature review. Furthermore I also discussed CDx types according to the molecular diagnostic technology such as immunohistochemistry (IHC), polymerase chain reaction (PCR), in situ hybridization (ISH), and next-generation sequencing (NGS) not only in the approved CDx but also in the developing one by US FDA. And I suggested the technology issue of CDx development process such as a selection of molecular diagnostics at the time of release, a clear understanding of the CDx mechanism, and a convergence of drug with CDx development. The necessity of social insurance system also was proposed for CDx development.

Pseudogenes: Nuances and Nuisances in Molecular Diagnostics

  • Oh, Seung Hwan
    • Journal of Interdisciplinary Genomics
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    • v.4 no.2
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    • pp.19-23
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    • 2022
  • Pseudogenes are genomic regions that contain gene-like sequences that have a high similarity to the known genes but are nonfunctional. They are categorized into processed, unprocessed, and unitary pseudogenes. Unprocessed pseudogenes generated by duplications can be problematic in sequencing approaches in molecular diagnostics. We discuss the risk of misdiagnosis when investigating genes with pseudogenes of high homology, and describe a method for identifying these small and annoying differences between parent genes and pseudogenes, including parent gene-specific assay design.

Orphan Nuclear Receptor Nurr1 as a Potential Novel Marker for Progression in Human Prostate Cancer

  • Wang, Jian;Yang, Jing;Zou, Ying;Huang, Guo-Liang;He, Zhi-Wei
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.3
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    • pp.2023-2028
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    • 2013
  • A number of studies have indicated that Nurr1, which belongs to a novel class of orphan nuclear receptors (the NR4A family), is important for carcinogenesis. Here we investigated expression of Nurr1 protein in benign and malignant human prostate tissues and association with clinicopathologic features using immunohistochemical techniques. Moreover, we also investigated the ability of Nurr1 to influence proliferation, migration, invasion and apoptosis of human prostate cancer cells using small interfering RNA silencing. Immunohistochemical analysis revealed that the expression of Nurr1 protein was higher in prostate cancer tissues than in benign prostate tissue (P<0.001), levels being positively correlated with tumor T classification (P = 0.003), N classification (P = 0.017), M classification (P = 0.011) and the Gleason score (P = 0.020) of prostate cancer patients. In vitro, silencing of endogenous Nurr1 attenuated cell proliferation, migration and invasion, and induced apoptosis of prostate cancer cells. These results suggest that Nurr1 may be used as an indicator for prostate cancer progression and be useful for novel potential therapeutic strategies.

Clinicopathological Significance of S100A10 Expression in Lung Adenocarcinomas

  • Katono, Ken;Sato, Yuichi;Jiang, Shi-Xu;Kobayashi, Makoto;Saito, Keita;Nagashio, Ryo;Ryuge, Shinichiro;Satoh, Yukitoshi;Saegusa, Makoto;Masuda, Noriyuki
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.1
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    • pp.289-294
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    • 2016
  • Background: S100A10, of the S100 protein family, is reported to be involved in cancer cell invasion and metastasis. The aims of the present study were to immunohistochemically examine S100A10 expression in surgically resected lung adenocarcinomas, and evaluate any relationships with clinicopathological parameters and prognosis of patients. Materials and Methods: S100A10 expression was immunohistochemically studied in 202 consecutive resected lung adenocarcinomas, and its associations with clinicopathological parameters were evaluated. Kaplan-Meier survival analysis and Cox proportional hazards models were used to estimate the effect of S100A10 expression on survival. Results: S100A10 expression was detected in 65 of the 202 (32.2%) lung adenocarcinomas, being significantly correlated with poorer differentiation (P =0.015), a higher pathological TNM stage (stages II and III) (P=0.004), more frequent and severe intratumoral vascular invasion (P=0.001), and a poorer prognosis (P=0.030). However, S100A10 expression was not an independent predictor of survival after controlling for clinicopathological factors. Conclusions: The present study reveals that S100A10 is expressed in a subset of lung adenocarcinomas, and this is related to some clinicopathological parameters, although further studies are required to confirm the correlation between S100A10 expression and prognosis of lung adenocarcinoma patients.

COVID-19: an update on diagnostic and therapeutic approaches

  • Iyer, Mahalaxmi;Jayaramayya, Kaavya;Subramaniam, Mohana Devi;Lee, Soo Bin;Dayem, Ahmed Abdal;Cho, Ssang-Goo;Vellingiri, Balachandar
    • BMB Reports
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    • v.53 no.4
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    • pp.191-205
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    • 2020
  • The unexpected pandemic set off by the novel coronavirus 2019 (COVID-19) has caused severe panic among people worldwide. COVID-19 has created havoc, and scientists and physicians are urged to test the efficiency and safety of drugs used to treat this disease. In such a pandemic situation, various steps have been taken by the government to control and prevent the Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2). This pandemic situation has forced scientists to rework strategies to combat infectious diseases through drugs, treatment, and control measures. COVID-19 treatment requires both limiting viral multiplication and neutralizing tissue damage induced by an inappropriate immune reaction. Currently, various diagnostic kits to test for COVID-19 are available, and repurposing therapeutics for COVID-19 has shown to be clinically effective. As the global demand for diagnostics and therapeutics continues to rise, it is essential to rapidly develop various algorithms to successfully identify and contain the virus. This review discusses the updates on specimens/samples, recent efficient diagnostics, and therapeutic approaches to control the disease and repurposed drugs mainly focusing on chloroquine/hydroxychloroquine and convalescent plasma (CP). More research is required for further understanding of the influence of diagnostics and therapeutic approaches to develop vaccines and drugs for COVID-19.