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Collection and Processing of Arterial Specimens for Histological Analysis

After atherosclerotic development has proceeded for the allotted time, arteries can be harvested by several techniques to optimize both quantitative and qualitative histological analysis. A study design must include consideration of the parameters to be analyzed, to determine ...

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Intravascular Ultrasound Imaging in the Quantitation of Atherosclerosis in Vivo

A number of imaging modalities have been used for evaluating the severity of atherosclerotic lesions in vivo. X-ray angiography, using iodine contrast agent, has been the standard imaging technique so far, in spite of its limitations. The severity of lumen-narrowing lesions is generally u ...

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Identification of Cell Types and Quantification of Lesion Composition

Many different cell types have been identified in atherosclerotic plaques, including endothelial cells, smooth muscle cells, monocytes, monocyte-derived macrophages, lymphocytes, and mast cells. Examining the role each cell type plays in the formation and pathophysiology of a ...

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1Nonradioactive In Situ Hybridization in Atherosclerotic Tissue

In situ hybridization (ISH) is a powerful and important technique that allows the detection and microscopic localization of nucleic acids within the specific cell, tissue, or chromosome of interest. In addition, it offers increased sensitivity over traditional filter hybridizat ...

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Evaluation of Angiogenesis

Every year, thousands of individuals around the world suffer from severe lower extremity vascular disorders that are amenable to neither medical nor surgical treatment, and thus amputation is required. A substantial portion of these amputations might be avoided by stimulating blood ...

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Animal Models of Diet-Induced Atherosclerosis

Animals models of atherosclerosis develop lesions either spontaneously or by interventions such as dietary, mechanical, chemical, or immunological induction. Animal models provide a means for studying the underlying mechanisms behind the atherosclerotic disease proces ...

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Gene Transfer to the Vasculature Historical Perspective and Implications for Future Research Objectives

Since the first demonstration of gene transfer to vascular tissue in 1990 (1), the field of gene therapy has grown at an astonishing pace. This is due to the lack of an effective and safe pharmacological treatment for certain vascular diseases and the availability of different gene transfer vehic ...

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A Pig Model of Vein Graft Disease Applications for Potential Gene Therapies

The development of an effective and safe gene therapy for prevention of vein graft failure, either acute or chronic, relies on the use of applicable and reproducible models of vein graft failure. Evaluation of potential new therapies usually involves assessment of beneficial phenotypic ...

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Mechanical Injury Models: Balloon Catheter Injury to Rat Common Carotid Artery

Removal of arterial endothelium and damage to medial smooth muscle with a balloon embolectomy catheter lead to formation of a thin mural thrombus, platelet adhesion and degranulation, smooth muscle cell migration to the intima, and cell proliferation and matrix synthesis, ultimately ...

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Lipoprotein Isolation and Analysis from Serum by Preparative Ultracentrifugation

Plasma lipoproteins are a heterogeneous population of soluble, macromolecular aggregates of lipids and proteins. They are responsible for the transport of waterinsoluble nutrients through the vascular and extravascular fluids from their site of synthesis or absorption to per ...

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Genetically Manipulated Models of Atherosclerosis in Mice

Mice are largely resistant to atherosclerosis. However, with dietary intervention or genetic manipulation, mice can be induced to develop atherosclerosis The focus of this chapter is genetically manipulated models (see Chapter 1 for discussion regarding diet-induced atheros ...

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Separation of Plasma Lipoproteins in Self-Generated Gradients of Iodixanol

The major classes of plasma lipoprotein, very low density lipoproteins (VLDL), low-density lipoproteins (LDL), and high-density lipoproteins (HDL), are characterized on the basis of differences in density and charge ( Table 1 ). Centrifugation is the ‘gold-standard’ for the analysis of p ...

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Fractionation of Lipoprotein Subclasses in Self-Generated Gradients of Iodixanol

Chapter 5 described the use of self-generated gradients of iodixanol for the fractionation of human plasma lipoproteins into the major classes: high-density, low-density, and very low density (HDL, LDL, and VLDL). During the metabolism of plasma HDL and LDL, the lipid and apoprotein composi ...

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7 Detecting Cholesteryl Ester Transfer Protein in Plasma

Transport of triglycerides (TGs) and cholesteryl esters (CEs) in plasma can be viewed as taking place via two major groups of lipoproteins, the TG-rich lipoproteins (chylomicrons and VLDL), on one hand, and the cholesterol-rich lipoproteins (LDL and HDL), on the other. The metabolism of these g ...

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8 Determination of Plasma Homocysteine

A number of epidemiological and clinical studies have linked elevated plasma homocysteine (Hcy) to atherosclerotic vascular disease affecting coronary, carotid, and peripheral vessels. Plasma Hcy can be considered a marker of methionine metabolic efficiency, mainly affect ...

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9 Assay for Serum Glycated Lipoproteins

Modified low density lipoprotein (LDL) is considered to be a risk factor for the development and progression of atherosclerosis (1-3). Glycated LDL and oxidized LDL are the only modified lipoproteins that exist naturally in the human body, but oxidized LDL does not exist in the circulation bec ...

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Culture of Immortalized Chondrocytes and Their Use As Models of Chondrocyte Function

Immortalization of chondrocytes increases life span and proliferative capacity but does not necessarily stabilize the differentiated phenotype. Expansion of chondrocyte cell lines in continuous monolayer culture may result in the loss of phenotype, particularly if high cell ...

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Immortalization of Human Articular Chondrocytes for Generation of Stable, Differentiated Cell Lines

Immortalized chondrocytes of human origin have been developed to serve as reproducible models for studying chondrocyte function. In this chapter, methods for immortalization of primary human chondrocytes with SV40-TAg, HPV-16 E6/E7, and telomerase by retrovirally mediated tr ...

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Generation of Pluripotent Stem Cells and Their Differentiation to the Chondrocytic Phenotype

It is well documented that adult cartilage has minimal self-repair ability. Current methods for treatment of cartilage injury focus on the relief of pain and inflammation and have met with limited long-term success. In the forefront of new therapeutic approaches, autologous chondrocy ...

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Semiquantitative Analysis of Gene Expression in Cultured Chondrocytes by RT-PCR

Reverse transcriptase-polymerase chain reaction (RT-PCR) is a powerful, sensitive, and rapid method to monitor small amounts of nucleic acids. This is of particular interest for small amounts of cells, as in cartilage. We present here two protocols to isolate total RNA and a protocol to study ma ...

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