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After inserting the needle, hold it steady for 35 seconds before withdrawal to ensure the full dose is delivered into subcutaneous tissue

Quantitative results from pre-acute, acute, remission and relapse samples were checked for normality and analyzed using one-way ANOVA with Tukeys correction for multiple comparisons, or with the Kruskal-Wallis test followed by Dunns multiple comparison test

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subsequently, the supernatant was titrated with DCPIP and the absorbance at 540 nm was measured by using a microplate reader infinite M1000 (Tecan Ltd.)