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For liver tissue preparation, blocks from each rat were frozen in liquid nitrogen and stored at −80°C in order to analyze IL-6 and TNFα.
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The paraffin-embedded ankle joints from each rat were sectioned, and endogenous peroxidase was blocked with 0.3% H2O2 for 10 minutes.
The organs including the liver, kidney, spleen, testis, and brain were collected from each rat, fixed with 10 wt.% formaldehyde solution at room temperature, embedded in paraffin block, sectioned, followed by staining with hematoxylin and eosin (H&E).
Bilateral rVLM tissue samples were collected from each rat.
Baseline images were obtained from each rat before treatment was administered.
One thin section was prepared from each rat.
After 1, 3 and 5 days, 10 rats from the control group and 10 rats from each time group of the experimental group were sacrificed, and both left and right kidneys from each rat were removed separately.
Several piriform cortex slices were obtained from each rat.
Five readings were obtained from each rat.
Liver tissue samples from each rat were fixed in Bouin's solution for 24 h, then dehydrated in a graded series of isopropyl alcohol, and embedded in paraffin blocks.
Two such slides were performed from each rat.
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