Medicine of the Future in America

Interferon-alpha-induced changes in metallothionein expression: Methods (Part 4)

One pathologist categorized the staining as rare (less than 5% of the cells stained), mild (5% to less than 30%), moderate (30% to less than 60%) or strong (more than 60% of the cells stained) and scored as follows: rare = 1, mild = 2, moderate = 3 or strong = 4. Standards for each grade of MT staining and MT-mRNA expression are shown in Figures 1 and 2.

PCNA labelling index

Liver specimens were allowed to react with the first antibody, a 1:200 dilution of anti-PCNA monoclonal antibody (DAKO Japan Co Ltd, Japan), at 4°C for 12 h, and then with biotin-labelled antirabbit immunoglobulin G antibody and horseradish peroxidase-bound EnVision (DAKO Japan Co Ltd, Japan). The specimens were subjected to 3,3′-diaminobenzidine reaction by the addition of 0.3% hydrogen peroxide solution and nucleus staining with Mayer’s hematoxylin, dehydrated and mounted. The proportion of positive cells per 400 liver cells was calculated as the PCNA labelling index. Please make sure you read all the information provided above to choose the right medications at the my canadian family pharmacy to benefit the most from the treatment. At this pharmacy, you will find tons of cheap and efficient meds for any problem.

Statistical analysis

Data are expressed as mean ± SD. Statistical analyses were performed with the StatView-J4.5 computer program (Abacus Concepts Inc, USA). Differences in HAI score, MT mRNA score, and PCNA labelling index before and after IFN therapy were analyzed with Wilcoxon’s signed rank test. Nonparametric data were compared using the Mann-Whitney U test. All P-values were twotailed, and P<0.05 was regarded as statistically significant.

The criterion of each grade of metallothionein staining

Figure 1) The criterion of each grade of metallothionein staining (immunohistochemical stain, original magnification x200). Arrows indicate metallothionein-positive hepatocytes. A Rare; B Mild; C Moderate; D Strong 

The criterion of each grade of metallothionein messenger RNA expression

Figure 2) The criterion of each grade of metallothionein messenger RNA expression (in situ hybridization, original magnification x200). Arrows indicate metallothionein messenger RNA-positive hepatocytes. A Rare; B Mild; C Moderate; D Strong

This entry was posted in Interferon-alpha and tagged Chronic hepatitis C, Interferon-alpha, Knodell histological activity index score, Metallothionein, PCNA labelling index.
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