Vaccination is a safe and effective approach to prevent deadly diseases. To increase vaccine production, we propose that a mechanical stimulation can enhance protein production. In order to prove this hypothesis, Sf9 insect cells were... more
Vaccination is a safe and effective approach to prevent deadly diseases. To increase vaccine production, we propose that a mechanical stimulation can enhance protein production. In order to prove this hypothesis, Sf9 insect cells were used to evaluate the increase in the expression of a fusion protein from hepatitis B virus (HBV S1/S2). We discovered that the ultrasound stimulation at a frequency of 1.5 MHz, intensity of 60 mW/cm2, for a duration of 10 minutes per day increased HBV S1/S2 by 27%. We further derived a model for transport through a cell membrane under the effect of ultrasound waves, tested the key assumptions of the model through a molecular dynamics simulation package, NAMD (Nanoscale Molecular Dynamics program) and utilized CHARMM force field in a steered molecular dynamics environment. The results show that ultrasound waves can increase cell permeability, which, in turn, can enhance nutrient / waste exchange thus leading to enhanced vaccine production. This finding ...
Research Interests: Engineering, Microbiology, Biophysics, Materials Science, Molecular Biology, and 15 moreComputational Biology, Biotechnology, Cell Biology, Infectious Diseases, Medicine, Molecular Dynamics, Model, HBV, Glucose, Animals, Insects, Hepatitis B virus, Cell Membranes, Hepatitis B Vaccines, and Cell membrane permeability
Purpose: MicroRNAs (miRNAs) post-transcriptionally regulate hundreds of gene targets involved in tumorigenesis thereby controlling vital biological processes, including cellular proliferation, differentiation and apoptosis. MiRNA... more
Purpose: MicroRNAs (miRNAs) post-transcriptionally regulate hundreds of gene targets involved in tumorigenesis thereby controlling vital biological processes, including cellular proliferation, differentiation and apoptosis. MiRNA profiling is an emerging tool for the potential early detection of a variety of malignancies. This study was conducyed to assess the feasibility and methodological robustness of quantifying sputum miRNAs, employing quantitative real-time polymerase chain reaction (RT-qPCR) and cluster analysis on an optimized miRNA profile as a novel approach for the early detection of non-small cell lung cancer (NSCLC). Methods: The relative expressions of 11 miRNAs in sputum (miR-21, miR-145, miR-155, miR-205, miR-210, miR-92, miR-17-5p, miR-143, miR-182, miR-372, and let-7a) in addition to U6 were retrospectively assessed in four NSCLC-positive and four negative controls. Subsequently, a set of five miRNAs (miR-21, miR-143, miR-155, miR-210, miR-372) was selected because...
Research Interests: MicroRNA, Biology, Medicine, Lung Cancer, Prospective studies, and 15 moreHumans, Female, Male, Aged, Middle Aged, microRNAs, Adult, Retrospective Studies, Sputum, Sensitivity and Specificity, Gene expression profiling, real time polymerase chain reaction, Early detection of cancer, Medical and Health Sciences, and lung neoplasms
Purpose: Nanotechnology is an emerging field with significant translational potential in medicine. In this study, we applied gold nanoparticles (GNP) to enhance radiation sensitivity and growth inhibition in radiation-resistant human... more
Purpose: Nanotechnology is an emerging field with significant translational potential in medicine. In this study, we applied gold nanoparticles (GNP) to enhance radiation sensitivity and growth inhibition in radiation-resistant human prostate cancer cells. Methods: Gold nanoparticles (GNPs) were synthesized using HAuCl4 as the gold particle source and NaBH4 as the reductant. Either thio-glucose or sodium citrate was then added to the solution separately to bind the GNPs to form thio-glucose-capped gold nanoparticles (Glu-GNP) and neutral gold nanoparticles (TGS-GNPs). Human prostate carcinoma DU-145 cells were exposed to vehicle, irradiation, 15nM TGS-GNPs, or 15nM Glu-GNPs, or GNPs plus irradiation. The uptake assays of GNP were performed using hemocytometer to count cells and the mass spectrometry was applied to calculate gold mass. The cytotoxicity induced by GNPs, irradiation, or GNPs plus irradiation was measured using a standard colorimetric MTT assay. Results: Exposure to Glu...
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Purpose: Small non-coding microRNAs (miRNAs) are key components of cancer development and are considered as potential biomarkers for cancer diagnosis and treatment monitoring. This study investigated miRNA expression profiles of human... more
Purpose: Small non-coding microRNAs (miRNAs) are key components of cancer development and are considered as potential biomarkers for cancer diagnosis and treatment monitoring. This study investigated miRNA expression profiles of human cancer cells in order to develop a screening method for lung cancer. Methods: A series of lung cancer related miRNAs (miR-21, miR-145, miR-155, miR-205, miR-210, miR-92, miR-17-5p, miR-143, miR-182, miR-372, let-7a) were selected as candidates for miRNA expression profiles of human lung cancer cell lines (A549, SK-mes-1). MicroRNA u6 was the endogenous control. Cancer cell lines for positive controls; breast MCF-7, prostate Du-145, and glioblastoma U118. The negative control was normal lung fibroblast cell line MRC-5. RT-PCR was performed on StepOnePlus (Applied Biosystem, USA). MiRNA expressions of malignant cells were compared with normal fibroblast cells as well as endogenous control (u6) using the thermal cycle at threshold. Assessment of miRNA exp...
Research Interests: MicroRNA, Cancer, Biology, Prostate Cancer, Cancer Research, and 13 moreMedicine, Lung Cancer, Humans, Female, Male, Cluster Analysis, microRNAs, Neoplasms, Reproducibility of Results, Gene expression profiling, Down-Regulation, reverse transcriptase polymerase chain reaction, and Medical and Health Sciences
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Hepatitis B is an infectious liver disease and vaccination is an effective way to protect individuals. We have applied mechanical wave stimulation to increase protein production. To validate our design, we used Sf9 insect cells to... more
Hepatitis B is an infectious liver disease and vaccination is an effective way to protect individuals. We have applied mechanical wave stimulation to increase protein production. To validate our design, we used Sf9 insect cells to increase antigen fragment fusion protein expression for hepatitis B virus (HBV S1/S2). We discovered that stimulation at a frequency of 1.5MHz, intensity of 60mW/cm(2), for a duration of 10 minutes per day increased HBV S1/S2 production by 15%. This finding is very significant for shortening vaccine production time or increasing the yield of proteins for use as vaccines.
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Research Interests: Chemistry, Molecular Biology, Fluorescence, Fluorescence Microscopy, Capillary electrophoresis, and 15 moreDNA damage, Medicine, DNA, Methods, Humans, Lasers, Mice, Animals, Laser Induced Fluorescence, Clinical Sciences, Time Factors, Immunoassay, Ionizing Radiation, immunoglobulin G, and fluorescent dyes
Sonodyanmic Therapy (SDT) is a new type of cancer therapy and has attracted lots of research attention recently. In this article, we present how nano-formulation helped SL052 to form a water soluble sono/photo-sensitize nanoparticle... more
Sonodyanmic Therapy (SDT) is a new type of cancer therapy and has attracted lots of research attention recently. In this article, we present how nano-formulation helped SL052 to form a water soluble sono/photo-sensitize nanoparticle (SL052-NPs) for cancer treatment. The nanostructure formation encapsulates SL052 and greatly improves the SL052 physicochemical properties without modifying its chemical structure. Nano-formulation also helps SL052 to
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Research Interests: Biology, Cytotoxicity, Medicine, Toxicity, Mice, and 15 moreAnimals, Cell Viability, Coefficient of Variation, Microelectrodes, Prediction Model, Real Time, Real Time Monitoring, Sensitivity and Specificity, Acute Toxicity, Cell Survival, Viability assay, Neutral Red, Automatic Detection, Cell count, and Pharmacology and pharmaceutical sciences
The toxic and mutagenic effects of ionizing radiation are believed to be caused by damage to cellular DNA. We have made use of a novel immunoassay for thymine glycol to examine the removal of this lesion from the DNA of irradiated human... more
The toxic and mutagenic effects of ionizing radiation are believed to be caused by damage to cellular DNA. We have made use of a novel immunoassay for thymine glycol to examine the removal of this lesion from the DNA of irradiated human cells. Because of the sensitivity of the assay, we have been able to keep the radiation doses at or below the standard clinical dose of 2 Gy. Our initial observations indicated that although removal of thymine glycol is > 80% complete by 4 h post-irradiation with 2 Gy, there is a lag of 30-60 min before repair commences. However, if cells are irradiated with 0.25 Gy 4 h prior to the 2-Gy dose, removal of the thymine glycols commences immediately after the second irradiation, suggesting that repair of thymine glycol is inducible. Our current studies are directed at two aspects of the repair process, (1) factors involved in the repair process leading up to and including glycosylase-mediated removal of thymine glycol and (2) the control of the inducible response. We have observed that mutation of the XPG gene drastically reduced the level and rate of global removal of thymine glycol (induced by 2-Gy irradiation), and there was no evidence for an inducible response. Similar results were seen with a Cockayne syndrome B (CSB) cell line. We have also examined repair in quiescent and phytohemagglutinin-stimulated human lymphocytes. Both show similar kinetics for the rate of removal of thymine glycol under induced and noninduced conditions.
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ABSTRACT In this paper, we explore the possibility of increasing the transfection rate of hard-to-transfect cells (such as KG-1 cells) using ultrasound-mediated gene delivery (USD). Single application ultrasound induced as well as the... more
ABSTRACT In this paper, we explore the possibility of increasing the transfection rate of hard-to-transfect cells (such as KG-1 cells) using ultrasound-mediated gene delivery (USD). Single application ultrasound induced as well as the synergistic properties of ultrasound with commonly used transfection reagents and repeated sessions USD were investigated and evaluated using GFP plasmid. KG-1 cells were not transfected by combining a commonly used transfection reagent and USD. However, repeated ultrasound-mediated treatments were able to increase transfection rate to 15% with a viability of 97%, as compared to 2% with the transfection reagent, Lipofectamine, alone.
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In this paper, we developed a low-cost intracellular delivery system based on microbubble and high gravity field. We successfully delivered FITC-Dextran (40kD) into hard-to-deliver THP-1 cells. The results showed that our method achieved... more
In this paper, we developed a low-cost intracellular delivery system based on microbubble and high gravity field. We successfully delivered FITC-Dextran (40kD) into hard-to-deliver THP-1 cells. The results showed that our method achieved high delivery efficiency up to 80%. It was found that the delivery efficiency and cell viability were closely related to the centrifuge speed. We speculated that the burst of microbubbles causes transient pore opening thus increasing the chance of biomolecules entering cells. This fast, low-cost and easy-to-operate protocol is very promising for delivering therapeutic genes and drugs into any cells which do not actively take up extracellular materials. This method is most effective for in-vitro delivery, but after delivery, treated cells might be injected back to human for in-vivo imaging.
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The present study explored the cellular uptake dynamics, the subcellular location and the internalization mechanisms of gold nanoparticles (GNPs) and glucose-capped GNPs (Glu-GNPs). The cancer radiotherapy-enhancing effects of GNPs were... more
The present study explored the cellular uptake dynamics, the subcellular location and the internalization mechanisms of gold nanoparticles (GNPs) and glucose-capped GNPs (Glu-GNPs). The cancer radiotherapy-enhancing effects of GNPs were also evaluated. We synthesized the GNPs and Glu-GNPs by the seeding technique. The effects on cellular uptake and the radiosensitizing effect induced by GNPs and Glu-GNPs at lower doses were investigated using two human cancer cell lines (HeLa and MCF-7). The intracellular location of the nanoparticles was analyzed by transmission electron microscopy (TEM). Analysis of cellular apoptosis following GNP-based radiotherapy was performed by flow cytometry and TUNEL assay. Cancer cells took up more Glu-GNPs than naked GNPs and the uptake curve showed size- and cell-dependent uptake. GNPs were mainly located in the cytoplasm and endocytosis is the mechanism behind the internalization of GNPs and Glu-GNPs. Lower doses of GNPs and Glu-GNPs still enhanced the...
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Research Interests: Technology, Nonparametric Statistics, Stem Cells, Biology, Medicine, and 14 moreBiological Sciences, Hematopoietic Stem Cells, Humans, Genetic Enhancement, Liposomes, Chemotaxis, Haematopoiesis, Gene Delivery, Transfection, Recombinant Proteins, Cell Survival, Hematopoietic Stem Cell Transplantation, Genetic Therapy, and Medical and Health Sciences
An ultrasensitive assay for measuring DNA base damage is described that couples immunochemical recognition with capillary electrophoresis and laser-induced fluorescence detection. The method provides a detection limit of 3 × 10 −21 moles,... more
An ultrasensitive assay for measuring DNA base damage is described that couples immunochemical recognition with capillary electrophoresis and laser-induced fluorescence detection. The method provides a detection limit of 3 × 10 −21 moles, an improvement of four to five orders of magnitude over current methods. Induction and repair of thymine glycols were studied in irradiated A549 cells (a human lung carcinoma cell line). Exposure of these cells to a low dose of radiation (0.25 Gray) 4 hours before a clinically relevant dose (2 Gray) enhanced removal of thymine glycols after the higher dose. These data provide evidence for an inducible repair response for radiation-induced damage to DNA bases.
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Research Interests: Materials Science, Flow Cytometry, Carbon Nanotube, Cell Biology, Nanotechnology, and 15 moreMedicine, Multidisciplinary, Hematopoietic Stem Cells, Cell Differentiation, Humans, Endocytosis, Cell Viability, Haematopoiesis, Gene Delivery, In Vivo, Magnetics, Cell Survival, Nanotubes Carbon, Adverse effect, and Cell count
Research Interests: Biophysics, Materials Science, Flow Cytometry, Fluorescence, Carbon Nanotube, and 15 moreNanotechnology, Cytotoxicity, Medicine, Multidisciplinary, DNA, Cell line, Humans, Gene transfer techniques, Cell Death, Ethanol, Magnetics, Chemical Precipitation, Biomolecule, Breast Cancer Cells, and Fluorescein
A microelectronic array assay was developed to specifically genotype Helicobacter pylori versus Helicobacter heilmannii and to determine antimicrobial resistance. Helicobacter 16S rRNA and 23S rRNA genes were specifically generated with... more
A microelectronic array assay was developed to specifically genotype Helicobacter pylori versus Helicobacter heilmannii and to determine antimicrobial resistance. Helicobacter 16S rRNA and 23S rRNA genes were specifically generated with Helicobacter genus-specific primers, respectively. The single-nucleotide polymorphisms (SNPs) in 16S rRNA, 268T specific in the H. pylori sequence, and 263A specific in H. heilmannii were used as molecular markers for identification of H. pylori and H. heilmannii, respectively. A triple-base-pair resistant mutation, AGA965-967TTC in 16S rRNA, is known to be responsible for H. pylori tetracycline resistance and was detected to identify resistant strains. H. pylori macrolide resistance was determined by the identification of 3 defined mutations in the 23S rRNA gene using the same method. The assay could be directly used to detect H. pylori in feces. The assay performs multiple determinations, including identification of Helicobacter species and antibio...
Research Interests: Microbiology, Biology, Helicobacter pylori, Medicine, Mutation, and 12 moreribosomal RNA, Electrophoresis, Genotype, Single Nucleotide Polymorphism, Feces, Genotyping, Microbial Sensitivity Tests, Biomolecular, Base Sequence, Biochemistry and cell biology, Molecular Sequence Data, and Tetracycline resistance
Research Interests: Library Science, Biology, Applied microbiology, Cytotoxicity, Medicine, and 14 moreMultidisciplinary, Cercopithecus aethiops, Humans, Escherichia coli, Animals, Feces, Laboratory Automation, Sensitivity and Specificity, Cell Survival, Vero cells, Crystal Violet, Viability assay, Biosensing Techniques, and VTEC
Magnetic gold nanoparticles (mGNPs) with uniform size and morphology synthesized by our sonication treatment method were covalently bound with fluorescein isothiocyanate (FITC) molecules. Driven by an external magnetic field,... more
Magnetic gold nanoparticles (mGNPs) with uniform size and morphology synthesized by our sonication treatment method were covalently bound with fluorescein isothiocyanate (FITC) molecules. Driven by an external magnetic field, FITC-labelled nanoparticles were delivered into plant cells with and without cell walls, evident from sectional transmission electron microscopy images. Confocal images further indicate that the green fluorescence in canola protoplasts and walled cells indeed came from the FITC molecules, instead of the chloroplasts’ autofluorescence. FITC-labelled nanoparticles had a delivery efficiency of 95% based on confocal images. In further study, plasmids were covalently bound with mGNPs, and delivered into canola cells with and without cell walls. After culturing for 48 h followed by staining with 5-bromo-4-chloro-3-indolyl-β-d-glucuronic acid (X-Gluc), blue colour appeared in the protoplasts, while the walled canola cells showed a green colour that can be interpreted ...
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Research Interests: Engineering, Chemistry, Treatment Outcome, Medicine, Biological Sciences, and 15 moreCell line, In Vitro, Humans, Mice, Animals, Real Time, CHEMICAL SCIENCES, In Vivo, Neoplasms, Analytical and Bioanalytical Chemistry, Cell Proliferation, Radiosensitivity, Cell Survival, Radiation Tolerance, and Clonogenic Assay
One aspect of gene therapy is the efficient transfection of a cell population with DNAs or RNAs (including siRNAs) in order to modulate the expression of genes, to produce cell types for use in gene therapy, or to meet other very... more
One aspect of gene therapy is the efficient transfection of a cell population with DNAs or RNAs (including siRNAs) in order to modulate the expression of genes, to produce cell types for use in gene therapy, or to meet other very important unmet medical needs. Gene therapy has a huge potential in modifying the blueprints of a human cell and