Improving the Efficacy of Conventional Therapy by Adding Andrographolide Sulfonate in the Treatment of Severe Hand, Foot, and Mouth Disease: A Randomized Controlled Trial

Background. Herb-derived compound andrographolide sulfonate (called Xiyanping injection) recommended control measure for severe hand, foot, and mouth disease (HFMD) by the Ministry of Health (China) during the 2010 epidemic. However, there is a lack of good quality evidence directly comparing the efficacy of Andrographolide Sulfonate combination therapy with conventional therapy. Methods. 230 patients were randomly assigned to 7–10 days of Andrographolide Sulfonate 5–10 mg/Kg/day and conventional therapy, or conventional therapy alone. Results. The major complications occurred less often after Andrographolide Sulfonate (2.6% versus 12.1%; risk difference [RD], 0.94; 95% CI, 0.28–1.61; P=0.006). Median fever clearance times were 96 hours (CI, 80 to 126) for conventional therapy recipients and 48 hours (CI, 36 to 54) for Andrographolide Sulfonate combination-treated patients (χ2=16.57, P<0.001). The two groups did not differ in terms of HFMD-cause mortality (P=1.00) and duration of hospitalization (P=0.70). There was one death in conventional therapy group. No important adverse event was found in Andrographolide Sulfonate combination therapy group. Conclusions. The addition of Andrographolide Sulfonate to conventional therapy reduced the occurrence of major complications, fever clearance time, and the healing time of typical skin or oral mucosa lesions in children with severe HFMD

Effects of MSK1 gene silencing and Thr-581 mutation on cytokine production in LPS-treated astrocytes.

<p><b>A</b>. Effects of siRNA for MSK1 and non-specific siRNA on LPS-induced expression of MSK1, p-MSK1 (Thr581) and iNOS were detected by Western blotting. <b>B</b>. The bar chart shows the ratio of total MSK1 and p-MSK1 to β-actin. <b>C–E</b>. ELISA showed that MSK1 gene silencing by siRNA further promoted the LPS-mediated upregulation of inflammatory cytokines. <b>F</b>. Western blot analysis showed the effect of mutation of Thr-581 to an alanine residue on LPS-induced expression of p-MSK1 Thr-581, p-MSK1 Ser-360, and total MSK1. <b>G</b>. The bar chart shows the ratio of p-MSK1 Thr-581 and p-MSK1 Ser-360 to total MSK1. <b>H–J</b>. ELISA showed the effect of mutation of Thr-581 on LPS-induced TNFα, IL-6, and IL1-β production in activated astrocytes.</p

Expression profile of MSK1 and p-MSK1 (Thr-581 and Ser-360) following LPS intracerebral injection.

<p><b>A</b>. Protein levels of t-MSK1, p-MSK1 Thr-581, p-MSK1 Ser-360 were detected before (control) and after injury. GAPDH was also detected by Western blotting. <b>B</b>. Quantification graphs (relative optical density) of the intensity of staining of p-MSK1 (Thr-581) and total MSK1 to GAPDH at each time point. GAPDH was used to confirm that equal amounts of protein were run on the gel. <b>C–H</b>. Immunofluorescence staining of MSK1 and p-MSk1 (Thr581) was performed to assess the staining changes for MSK1 and p-MSK1 immunoreactivity in the cortex at day 1 after LPS-injection. <b>I</b>. Negative control. * and ^# indicate significant differences at P<0.05, compared with normal brain cortex. Scale bars: 40 µm (C–F), 20 µm (G–J).</p

Immunolocalization of MSK1 and p-MSK1 (Thr-581) with different cellular markers in cerebral cortex by double immunofluorescence staining.

<p>In the adult rat brain cortex, within 5(red, <b>A</b> and <b>E</b>) and p-MSK1 (Thr581) (red, <b>I</b> and <b>M</b>) and different cell markers (green, <b>B, F, J, N</b>), such as a neuronal marker (NeuN) and an astrocyte marker (GFAP). The yellow color in the merged images represents colocalization of MSK1 or p-MSK1 (Thr581) with different phenotype-specific markers (<b>C, G, K, O</b>). Colocalization of MSK1 and p-MSK1 (Thr581) with different phenotype-specific markers in the normal group are shown in the brain cortex (<b>D, H, L, P</b>). Quantitative analysis of different phenotype-specific marker-positive cells expressing MSK1 (<b>Q</b>) and p-MSK1 (<b>R</b>) (%) in the unit area (mm²) in the normal group and 1 day after injury. *indicates significant difference at P<0.05, compared with the normal group. Error bars indicate SEM. Scale bars: 20 µm (<b>A–P</b>).</p