ABSTRACT
Background@#Brucella infection induces brucellosis, a zoonotic disease. The intracellular circulation process and virulence of Brucella mainly depend on its type IV secretion system (T4SS) expressing secretory effectors. Secreted protein BspJ is a nucleomodulin of Brucella that invades the host cell nucleus. BspJ mediates host energy synthesis and apoptosis through interaction with proteins. However, the mechanism of BspJ as it affects the intracellular survival of Brucella remains to be clarified. @*Objectives@#To verify the functions of nucleomodulin BspJ in Brucella's intracellular infection cycles. @*Methods@#Constructed Brucella abortus BspJ gene deletion strain (B. abortus ΔBspJ) and complement strain (B. abortus pBspJ) and studied their roles in the proliferation of Brucella both in vivo and in vitro. @*Results@#BspJ gene deletion reduced the survival and intracellular proliferation of Brucellaat the replicating Brucella-containing vacuoles (rBCV) stage. Compared with the parent strain, the colonization ability of the bacteria in mice was significantly reduced, causing less inflammatory infiltration and pathological damage. We also found that the knockout of BspJ altered the secretion of cytokines (interleukin [IL]-6, IL-1β, IL-10, tumor necrosis factor-α, interferon-γ) in host cells and in mice to affect the intracellular survival of Brucella. @*Conclusions@#BspJ is extremely important for the circulatory proliferation of Brucella in the host, and it may be involved in a previously unknown mechanism of Brucella's intracellular survival.
ABSTRACT
<p><b>OBJECTIVE</b>To develop a RP-HPLC method for simultaneous determination of liquiritin, naringin, hesperidin and glycyrrhizic acid in extraction of Wendan formula.</p><p><b>METHOD</b>DIKMA Diamonsil(2)-C18 column (4.6 mm x 250 mm, 5 microm) was used at 25 degrees C with the mobile phase of acetonitrile-0.1% phosphatic acid in a gradient manner. The flow rate was set at 1.0 mL min(-1). The detection wavelength was 237, 283 nm.</p><p><b>RESULT</b>The linear responses ranged from 0.0199-0.1191 microg for liquiritin (r = 0.9997, n = 6), 0.1800-1.0800 microg for naringin (r = 0.9997, n = 5), 0.1455-0.8730 microg for hesperidin (r = 0.9998, n = 6), 0.0393-0.2355 microg for monoammonium glycyrrhizinate (r = 0.9997, n = 6), respectively. The average recoveries were 97.7% with RSD 1.5% for liquiritin, 97.7% with RSD 2.0% for naringin, 97.1% with RSD 2.0% for hesperidin and 98.5% with RSD 1.9% for glycyrrhizic acid, respectively.</p><p><b>CONCLUSION</b>The method is quick, simple and repeatable for simultaneous determination of liquiritin, naringin, hesperidin and glycyrrhizic acid in extraction of Wendan formula.</p>