Manual immunohistochemistry for detecting ALK gene fusion antibody in 519 cases of non-small lung cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of anaplastic lymphoma kinase (ALK) gene fusion antibody in non-small cell lung cancer (NSCLC) and explore the clinicopathological significance.</p><p><b>METHODS</b>Using manual immunohistochemistry (IHC) with D5F3 rabbit monoclonal antibody, we detected the expression of ALK gene fusion protein in 519 cases of NSCLC. The relations of ALK fusion protein with the clinical characteristics of the patients and the histological classification of the tumors were analyzed. The expressions of ALK fusion protein were compared between surgical specimens and biopsy samples, and the consistency of manual IHC results was evaluated with the results of a fully automated IHC instrument and fluorescence in situ hybridization (FISH).</p><p><b>RESULTS</b>The positivity rate of ALK fusion protein was 11.37% (59/519) among the cases detected by manual IHC. The patients tended to have a young age of onset (P=0.048) and most of the tumors were adenocarcinoma. In the surgical specimens, ALK fusion protein was expressed mostly in invasive mucinous adenocarcinoma (P<0.01), and it was a high risk factor of lymph node metastasis [OR=2.188(95%C.I:1.161-4.122)]. No statistical difference was found in the test results of manual IHC between surgical specimens and biopsy samples. The results by manual IHC suggesting a strong expression were consistent with the results by automated IHC and FISH.</p><p><b>CONCLUSION</b>Manual IHC can be reliable for screening ALK fusion arrangement in patients with NSCLC.</p>

Effect of protein kinase on endothelial cytoskeleton induced by septic shock / 中华外科杂志

<p><b>OBJECTIVE</b>To study the effect of cGMP-dependent protein kinase (PKG) on the pathogenesis of septic shock.</p><p><b>METHODS</b>Confluent endothelial cells were disintegrated and centrifugated to obtain cell lysates after being treated with LPS or PKG activator 8-Br-cGMP. PKG activity of lysates was measured with radioactive isotope label method in a reaction system of phosphorylation of specific substrate H2B by PKG, and the shape and the distribution of intracellular filamentous actin were detected by specific fluorescence staining. For the control study, the PKG specific inhibitor KT5823 was used to pretreat the endothelial cells before the administration of LPS or PKG activator 8-Br-cGMP.</p><p><b>RESULTS</b>Exposure to LPS for 5, 10, 30 and 60 minutes led to a rapid time-dependent increase in endothelial PKG activity (P < 0.01 compared to the blank) and the polar distribution of intracellular filamentous actin and preincubation with KT5823 abolished these effects. 8-Br-cGMP was similar to LPS.</p><p><b>CONCLUSIONS</b>The results suggested that LPS can mediate PKG activation and the stress variety of filamentous actin in the vascular endothelial cells, which probably induce the endothelial hyperpermeability after septic shock.</p>