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Articles by Hong Zhang in JoVE

 JoVE Immunology and Infection

Isolation of Human Umbilical Vein Endothelial Cells and Their Use in the Study of Neutrophil Transmigration Under Flow Conditions


JoVE 4032 8/08/2012

Department of Physiology and Pharmacology, University of Calgary

This article first describes a procedure for isolating human endothelial cells from umbilical veins and then shows how to use these cells to examine neutrophil transmigration under flow conditions. By using a low-volume flow chamber made from a polymer with the optical characteristics of glass, live-cell fluorescent imaging of rare cell populations is also possible.

Other articles by Hong Zhang on PubMed

Genes Under Positive Selection in Mycobacterium Tuberculosis

We employed an evolutionary genomics approach to detect genes under lineage-specific positive selection for the two closely related Mycobacterium tuberculosis strains, the virulent H37Rv and the avirulent H37Ra, with the clinical isolate CDC1551 as the outgroup. We found six H37Rv-specific and six H37Ra-specific positively selected genes, among which the former comprised a flavoprotein, a RNA polymerase sigma factor SigM, two PPE family proteins, as well as two hypothetical proteins, while the latter consisted of a dehydrogenase, a (3R)-hydroxyacyl-ACP dehydratase subunit HadA, a PPE family protein, and three PE-PGRS family proteins. Obviously, the PE/PPE/PE-PGRS family proteins were the main targets of positive selection. The functional discussion of our findings implied that those positively selected genes were highly involved in antigen variations and immune evasions of Mycobacterium tuberculosis.

Preparation of PLGA Scaffolds with Graded Pores by Using a Gelatin-Microsphere Template As Porogen

Porous scaffolds with graded pores are crucial to osteochondral regeneration. In this study, a technique combining solution casting with gelatin-microsphere template leaching has been developed to produce poly(l-lactide-co-glycolide) (PLGA) scaffolds with graded pores. The traditional emulsification and solvent extraction method was improved by using the gradient ethanol/water solutions to extract water to prepare gelatin microspheres with a smooth surface without the use of any surfactant. Gelatin microspheres with different diameters were in sequence put into a custom-made cylindrical Teflon mold and bonded together to obtain gelatin-microsphere templates. By using the gelatin-microsphere templates as porogen, PLGA scaffolds with graded pore size across the cylindrical axis were prepared. The porosity of the scaffold was as high as 95%. The pore size effect on osteoblasts was studied. The results showed that the graded scaffolds possessed good biocompatibility for osteoblast growth. During the 14 days culture, the cell proliferation of all the three pore layers displayed the trend of increasing. The proliferation rate of the large pore layer was lower than the other two layers. However, the difference of alkaline phosphatase activity on the three pore layers was not statistically significant. We assumed that it was probably because of the hydrophobicity and the short culture time. It was demonstrated that gradient ethanol/water solutions provided a simple way to prepared gelatin microspheres. The graded scaffolds would provide potential application for osteochondral regeneration.

Immunoglobulin E and Mast Cell Proteases Are Potential Risk Factors of Human Pre-diabetes and Diabetes Mellitus

Recent studies have suggested that mast-cell activation and inflammation are important in obesity and diabetes. Plasma levels of mast cell proteases and the mast cell activator immunoglobulin E (IgE) may serve as novel inflammatory markers that associate with the risk of pre-diabetes and diabetes mellitus.

A Genome-wide Association Study in Han Chinese Identifies Multiple Susceptibility Loci for IgA Nephropathy

We performed a two-stage genome-wide association study of IgA nephropathy (IgAN) in Han Chinese, with 1,434 affected individuals (cases) and 4,270 controls in the discovery phase and follow-up of the top 61 SNPs in an additional 2,703 cases and 3,464 controls. We identified associations at 17p13 (rs3803800, P = 9.40 × 10(-11), OR = 1.21; rs4227, P = 4.31 × 10(-10), OR = 1.23) and 8p23 (rs2738048, P = 3.18 × 10(-14), OR = 0.79) that implicated the genes encoding tumor necrosis factor (TNFSF13) and α-defensin (DEFA) as susceptibility genes. In addition, we found multiple associations in the major histocompatibility complex (MHC) region (rs660895, P = 4.13 × 10(-20), OR = 1.34; rs1794275, P = 3.43 × 10(-13), OR = 1.30; rs2523946, P = 1.74 × 10(-11), OR = 1.21) and confirmed a previously reported association at 22q12 (rs12537, P = 1.17 × 10(-11), OR = 0.78). We also found that rs660895 was associated with clinical subtypes of IgAN (P = 0.003), proteinuria (P = 0.025) and IgA levels (P = 0.047). Our findings show that IgAN is associated with variants near genes involved in innate immunity and inflammation.

Crocin Inhibits Proliferation and Nucleic Acid Synthesis and Induces Apoptosis in the Human Tongue Squamous Cell Carcinoma Cell Line Tca8113

Background: Cancer chemoprevention is a proven effective strategy for oral squamous cell carcinoma (OSCC). The present study was designed to investigate the effects of crocin, a potential chemopreventive agent, on growth and DNA and RNA content in a human tongue squamous cell carcinoma cell line, Tca8113. Methods: Tca8113 cells were treated with crocin for 24, 48, 72, and 96 h at concentrations of 0.1, 0.2, 0.4, and 0.8 mM. Tumor cell viability was investigated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. In addition, Tca8113 cells were treated with 0.4 mM crocin and cytotoxic effects as an inducer of apoptosis were analyzed using flow cytometry. Furthermore, acridine orange (AO) staining and observation using laser scanning confocal microscopy (LSCM) were used to determine the effects of the drug on nucleic acid synthesis. Results: Crocin decreased Tca8113 cell viability and growth remarkably at 24, 48, 72, and 96 h, in a concentration-dependent manner (P<0.05). In addition, 0.4 mM crocin significantly induced both early and late apoptosis of Tca8113 cells. Moreover, the cellular DNA and RNA content was significantly downregulated by 0.4 mM crocin compared with the negative control (P<0.01). Conclusions: Our observations support the feasibility of applying crocin as a chemoprophylactic agent and treatment for OSCCs.

[Surveillance on the Etiology and Genetic Characteristics of Rabies in Hunan Province, from 2008 to 2009]

To analyze the etiology of rabies in Hunan province and the genetic characteristics of rabies N gene isolated from 2008 to 2009.

Lys(48)-linked TAK1 Polyubiquitination at Lysine-72 Downregulates TNFα-induced NF-κB Activation Via Mediating TAK1 Degradation

Protein kinases are important regulators of intracellular signal transduction pathways and play critical roles in diverse cellular processes. TAK1, a member of the MAPKKK family, is essential for TNFα-induced NF-κB activation. Phosphorylation and Lys(63)-linked polyubiquitination (polyUb) of TAK1 are critical for its activation. However, whether TAK1 is regulated by polyubiquitination-mediated protein degradation after its activation remains unknown. Here we report that TNFα induces TAK1 Lys(48) linked polyubiquitination and degradation at the later time course. Furthermore, we provide direct evidence that TAK1 is modified by Lys(48)-linked polyubiquitination at lysine-72 by mass spectrometry. A K72R point mutation on TAK1 abolishes TAK1 Lys(48)-linked polyubiquitination and enhances TAK1/TAB1 co-overexpression-induced NF-κB activation. As expected, TAK1 K72R mutation inhibits TNFα-induced Lys(48)-linked TAK1 polyubiquitination and degradation. TAK1 K72R mutant prolongs TNFα-induced NF-κB activation and enhances TNFα-induced IL-6 gene expression. Our findings demonstrate that TNFα induces Lys(48)-linked polyubiquitination of TAK1 at lysine-72 and this polyubiquitination-mediated TAK1 degradation plays a critical role in the downregulation of TNFα-induced NF-κB activation.

[Double Balloon Endoscopy in Diagnosis of Patients with Obscure Gastrointestinal Bleeding]

Objective: To assess the diagnostic value of double balloon endoscopy (DBE) for obscure gastrointestinal bleeding (OGIB) Methods: The data of 103 OGIB patients who underwent DBE from January 2007 to September 2010 in the First Affiliated Hospital, Zhejiang University School of Medicine were retrospectively analyzed. Results: DBE was successfully performed in all 103 patients without complications. Of 103 patients, 66(64.1 %) had positive DBE findings and 28 had surgery procedures(27.2 %). Ninety-four patients finally acquired positive diagnosis, including small intestine tumor(31.1 %), angiodysplasia(22.3 %), exulceratio simplex(9.7 %), Crohn's disease(6.8 %), diverticulum(4.9 %), abdominal purpure(4.9 %), etc. Lesions occurred more frequently in proximal small intestine than in distal small intestine (56.3 % Compared with 30.1 %, P<0.001). Conclusion: DBE is a safe, effective and reliable procedure for the diagnosis of OGIB.

Molecular Imaging in Tracking Tumor Stem-like Cells

Cancer remains a major public health problem in many countries. It was found to contain a subset of cancer stem cells (CSCs) that are capable of proliferation and self-renewal, and differentiation into various types of cancer cells. CSCs often display characteristics of chemotherapy resistance and radiotherapy resistance. Numerous putative biomarkers of CSCs are currently identified including CD133, CD44, CD24, ALDH (aldehyde dehydrogenase), and ABCG2. Interestingly, no single marker is exclusively expressed by CSCs. Thus, the various combinations of different biomarkers will be possible to identify CSCs, and considerable work is being done to recognize new ones. In order to demonstrate the mechanisms of resistance and response to therapy and predict the outcome as well as prognosis, the ways to track and identify CSCs will be extremely important. The technologies of molecular imaging will reveal mechanisms of cancer progression and provide visual targets for novel therapeutics. Limited studies were investigated on the detection of various types of CSCs by molecular imaging. Although the tracking of circulating CSCs is still hampered by technological challenges, personalized diagnosis and therapies of cancers are expected to be established based on increased understanding of molecular imaging of cancer stem-like cells biomarkers.

California Almond Shelf Life: Lipid Deterioration During Storage

The effects of storage conditions on the lipid deterioration in California almond nuts and sliced were studied. Natural whole almonds with or without polyethylene (PE) packaging and blanched whole almonds and sliced with PE packaging were stored in 10 different storage conditions which were combinations of different temperatures and relative humidity levels. The peroxide values (PVs), iodine values (IVs), and free fatty acids (FFAs) were monitored during the storage. The PVs in the natural samples did not change noticeably whereas the blanched samples changed greatly, indicating that skins may have played a significant role. The IVs decreased slightly in the 1st 150 d of storage and then leveled off. The slightly faster changes in IVs in the blanched samples coincided with the greater changes in PVs in the blanched samples. The natural samples exhibited much higher FFA levels than the blanched samples after storage. In general, FFA increased with increasing storage time, temperature, and humidity. Highest levels of FFA were observed in the samples stored at high temperature and high humidity. PRACTICAL APPLICATION: The results reported in this article provide useful information that almond producers and processors could use to develop their storage and transport processes.

Protective Role of 1,25(OH)2vitamin D3 in the Mucosal Injury and Epithelial Barrier Disruption in DSS-induced Acute Colitis in Mice

ABSTRACT:

[The Diagnostic Value of CT-guided Percutaneous Needle Lung Biopsy in Diffuse Parenchymal Lung Diseases]

This study was to evaluate the efficacy and limitation of CT-guided percutaneous cutting needle lung biopsy in the diagnosis of diffuse parenchymal lung diseases (DPLD).

[Study on the Effect of Grape Seed Proanthocyanidins on Increasing the Radiosensitivity for X-ray]

To investigate the effect of Grape Seed Proanthocyanidins (GSPs) on enhancing the radiosensitivity of human hepatic carcinoma cell line HepG2, human cervical cancer cell line Hela and human leukemia cell line K562 for X-ray in vitro.

Oncogenic MST1R Activity in Pancreatic and Gastric Cancer Represents a Valid Target of HSP90 Inhibitors

Aim: To evaluate the effects of HSP90 blockade by EC154 on the oncogenic receptor tyrosine kinase macrophage-stimulating 1 receptor (MST1R) in gastric and pancreatic cancer.

Genetic Diversity and Drug Susceptibility of Mycobacterium Tuberculosis Isolates from Zunyi, One of the Highest Incidence Rate Areas in China

M. tuberculosis isolates from Zunyi were found more diversified but clustered less frequently to Beijing family compared to other areas of China. These observations, on top of the fact that Zunyi area has a high prevalence of MDR-TB, support the notion that Beijing-family isolates may not be linked to MDR-TB.

Phase 2 Trial of "sandwich" L-asparaginase, Vincristine, and Prednisone Chemotherapy with Radiotherapy in Newly Diagnosed, Stage IE to IIE, Nasal Type, Extranodal Natural Killer/T-cell Lymphoma

Extranodal natural killer/T-cell lymphoma (ENKTL), nasal-type, is a distinct entity of lymphoid tissue. ENKTL is sensitive to radiotherapy (RT), but the prognosis is poorer than for other types of early lymphoma. The treatment schedule is controversial.

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