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A semi-quantitative evaluation of renal tissues was accomplished by scoring the degree of severity according to previously published criteria.[27] All sections of kidney samples were examined for tubular necrosis. Briefly, minimum of 50 proximal tubules associated with 50 glomeruli were examined for each slide and an average score was obtained. Severity of lesion was graded from 0 to 3 according to the percentage of the tubular involvement. Slides were examined and assigned for severity of changes selleck chemicals llc using scores on a scale in which (0) denotes no change; grade 1 �C changes affecting 50% of tubules (severe). Histopathological evaluation was performed on left kidney tissues. Paraffin-embedded specimens were cut into 5-mm thick sections and stained with hematoxylin and eosin and Masson's trichrome for examination under a light microscope (BH-2; Olympus, Tokyo, Japan). To evaluate leukocyte infiltration, the widening of interstitial spaces with focal leukocyte infiltration was assessed in five randomly chosen sections prepared from each kidney sample. For each section, the average number of leukocytes per 0.28 mm2 was calculated from these leukocyte-infiltrated foci using a high-power microscopic field (��400). To estimate the grade of interstitial fibrosis, the interstitial area that was stained green PI3K Inhibitor Library in vivo with Masson's trichrome was evaluated as a percentage of the total examined area in five randomly chosen sections prepared from each kidney sample using an image analyzer (Leica; Leica Micros Imaging Solutions, Cambridge, UK). For each section, interstitial space widening with focal leukocyte infiltration and interstitial fibrosis was assessed in high-power fields (��400) to quantify the results. The Banff classification Fossariinae of kidney pathology was used for scoring the degree of mononuclear cell infiltration and interstitial fibrosis. The score was graded from 0 to 3, depending on the severity of histological characteristics.[19,28] Statistical analyses Results of all groups were shown as mean values �� standard deviation. Statistical analyses of the histopathologic evaluation of the groups were carried out by the Chi-square test and biochemical data were analyzed by the one-way analysis of variance. The significance between two groups was determined by the Dunnett's multiple comparison tests, and P