The relationship of the expression of principal pattern recognition receptor with the pulmonary injury in abdominal infection-induced sepsis / 中华外科杂志

<p><b>OBJECTIVE</b>In order to investigate the evidence of the synergistic effects of bacterial components, to observe the relationship of the expression of lipopolysaccharide (LPS) receptors [CD14, Toll-like receptor 4 (TLR4), scavenger receptor (SR)], lipoprotein receptor (TLR2) and bacterial DNA receptor (TLR9) with pulmonary injury in abdominal infection-induced sepsis.</p><p><b>METHODS</b>30 mice were used and randomly divided into cecal ligation puncture (CLP) (n = 15) and sham (n = 15) groups. The animals were respectively sacrificed 8, 12 and 24 (each point n = 5) hours following CLP and sham CLP. The lungs were removed and immediately stored in liquid nitrogen for TLRs mRNA, tumor necrosis factor (TNF) alpha and myeloperoxidase (MPO) assay. To detect the expression of CD14, TLR4, SR, TLR2 and TLR9 mRNA by reverse-transcription polymerase chain reaction, to detect the TNF-alpha content of the lung tissue by enzyme-labeled immunosorbent assay, and to assay the MPO activity of the lung tissue spectrophotometer.</p><p><b>RESULTS</b>It was found that the expression of receptors for LPS, BLP and bacterial DNA in pulmonary tissues was markedly changed in CLP-induced sepsis, showing upregulation of CD14 mRNA (1.143 +/- 0.139, t = 0.022, P < 0.05), TLR2 mRNA (0.418 +/- 0.102, t = 0.021, P < 0.05), TLR4 mRNA (0.595 +/- 0.052, t = 0.0001, P < 0.01) and TLR9 mRNA (0.743 +/- 0.178, t = 0.0023, P < 0.01) at different degrees (P < 0.05 or P < 0.01) after postinjury 8 h, among which the expression of TLR9 mRNA kept increasing. The expression of SR mRNA (8 h: 0.659 +/- 0.159; 12 h: 0.429 +/- 0.061; 24 h: 0.300 +/- 0.045; t = 0.029, P < 0.05; t = 0.001, P < 0.01; t = 0.003, P < 0.01) showed continuous down-regulation.</p><p><b>CONCLUSION</b>There was a marked correlation between the changes of pattern-recognition receptor expression and the increases of MPO and TNF-alpha levels in pulmonary tissues.</p>

Effects of Shexiang injection on expression of CD54 of mononuclear cells and soluble vascular cell adhesion molecule-1 in patients with acute cerebral infarction / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the clinical efficacy of Shexiang Injection (SI) on inflammatory reaction in patients with acute cerebral infarction (ACI).</p><p><b>METHODS</b>Forty-two patients with ACI were randomly divided into two groups, 21 in each group. The control group treated with conventional therapy and the SI group treated with conventional therapy plus SI. Besides, 21 healthy people were arranged in the normal group for control. Expression of CD54 of mononuclear cell (MC-CD54) and serum level of soluble vascular cell adhesion molecule-1 (sVCAM-1) were determined and the clinical efficacy was observed dynamically before treatment and on the 7th, 14th day of the course.</p><p><b>RESULTS</b>Levels of MC-CD54 expression and sVCAM-1 in the ACI patients increased obviously (P < 0.01), reached the peak on the 7th day, and declined obviously on the 14th day in both groups, however, the lowering in the SI group was more significant than that in the control group (P < 0.01); and positive correlation was shown between these two indexes (P < 0.01). After treatment, score of neural defect was improved more significantly (P < 0.01), and the markedly effective and curative rate was higher in the SI group than those in the control group (P < 0.05), respectively.</p><p><b>CONCLUSION</b>SI could inhibit the subsequent inflammatory reaction, and thus improve the clinical efficacy of conventional therapy in treating patients with ACI.</p>

The effects of TNF alpha and IFN gamma on the expression of pattern recognition receptors on the surface of mouse alveolar macrophages / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the effects of tumor necrosis factor alpha (TNF alpha) and interferon gamma (IFN gamma) on the expression of pattern recognition receptors (PRRs) on the surface of mouse alveolar macrophages.</p><p><b>METHODS</b>Alveolar macrophages from mouse were cultured in DMEM supplemented with 10% (V/V) endotoxin-free calf serum. After the alveolar macrophages were stimulated with TNF alpha and IFN gamma (concentration, 20 ng/ml) for 3 h, 6 h and 12 h, the expression of PRRs, including cluster of differentiation 14 (CD14), scavenger receptor (SR), toll-like receptor 4 (TLR4), TLR2 and TLR9 mRNA and proteins were examined by RT-PCR and immunohistochemistry.</p><p><b>RESULTS</b>The expressions of CD14, TLR2 and TLR9 receptors, which were related with cellular activation, were up-regulated by the stimulation of TNF alpha and IFN gamma (P < 0.05), while SR, which was related with cellular defense action, was down-regulated (P < 0.05). Although the expression of TLR4 was up-regulated, there was no statistical significance (P > 0.05).</p><p><b>CONCLUSIONS</b>The cytokines such as TNF alpha and IFN gamma could also produce feedback regulation on the expression of PRRs at the levels of genes and proteins. Such regulation on the PRRs expression would be significant for further amplification of inflammation cascade and eventually leading to uncontrolled inflammation.</p>

Novel multi-probe RNase protection assay set for detection of endotoxin associated receptors gene expression / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To construct the multi-probe ribonuclease protection assay (RPA) template set to be used for detecting expression patterns of MD-2, TLR4, CD14 mRNAs in human peripheral blood mononuclear cells.</p><p><b>METHODS</b>The designed cDNA fragments of the three genes were generated by polymerase chain reaction (PCR) using specific primers and directionally cloned into EcoR I and Hind III sites of expression plasmid pSP72 containing the T7 promoter, the linearized plasmids was used as template to synthesize anti-sense RNA probes. Then we extracted total RNA from peripheral blood mononuclear cells (PBMC) and detected the dynamic expression patterns of the three genes with RPA method.</p><p><b>RESULTS</b>The proper sequence and orientation of the template set were confirmed by sequencing and the template set was successfully used to assay TLR4, MD-2 and CD14 mRNAs in human PBMC. The results showed that the three detected genes decreased transiently 1-3 hours after 100 ng/ml LPS stimulation.</p><p><b>CONCLUSIONS</b>These new RPA multi-probe set provided valuable tool for the simultaneous quantitative determination of expression of TLR4, CD14 and MD-2 mRNAs in both constitutive and inducible types.</p>