Integrated analysis of multi-omics data reveals T cell exhaustion in sepsis.

Background: Sepsis is a heterogeneous disease, therefore the single-gene-based biomarker is not sufficient to fully understand the disease. Higher-level biomarkers need to be explored to identify important pathways related to sepsis and evaluate their clinical significance. Methods: Gene Set Enrichment Analysis (GSEA) was used to analyze the sepsis transcriptome to obtain the pathway-level expression. Limma was used to identify differentially expressed pathways. Tumor IMmune Estimation Resource (TIMER) was applied to estimate immune cell abundance. The Spearman correlation coefficient was used to find the relationships between pathways and immune cell abundance. Methylation and single-cell transcriptome data were also employed to identify important pathway genes. Log-rank test was performed to test the prognostic significance of pathways for patient survival probability. DSigDB was used to mine candidate drugs based on pathways. PyMol was used for 3-D structure visualization. LigPlot was used to plot the 2-D pose view for receptor-ligand interaction. Results: Eighty-four KEGG pathways were differentially expressed in sepsis patients compared to healthy controls. Of those, 10 pathways were associated with 28-day survival. Some pathways were significantly correlated with immune cell abundance and five pathways could be used to distinguish between systemic inflammatory response syndrome (SIRS), bacterial sepsis, and viral sepsis with Area Under the Curve (AUC) above 0.80. Seven related drugs were screened using survival-related pathways. Conclusion: Sepsis-related pathways can be utilized for disease subtyping, diagnosis, prognosis, and drug screening.

[Effect of endotracheal tube with subglottic suction line to stop tracheostomy bleeding in critical patients].

OBJECTIVE: To observe the effect of endotracheal tube with subglottic suction line on tracheostomy bleeding in critical patients. METHODS: This procedure was utilized for 35 critical patients with tracheostomy bleeding. With the help of bronchoscope, the endotracheal tube with subglottic suction line was inserted during tracheostomy to 0.5-1.0 cm above the carina. Then aerocyst of the tube was inflated, vacuum was set up for continuous suction via suction port, and hemocoagulase was injected to the opening of tracheostomy. The airway hemorrhage and treatment results were recorded. RESULTS: Tracheostomy bleeding in 33 patients (94.29%) were stopped. The bleeding time ranged from 6 to 48 hours with an average of (24.5±4.1) hours. The tube was in place for 2-10 days with a medium of 4 days. CONCLUSIONS: Using endotracheal tube with subglottic suction line for critical patients with tracheostomy bleeding is very effective. This procedure is also very simple and fast, and is best suitable for patient with heavy bleeding in tracheostomy.

Clinical evaluation of a homemade enzyme-linked immunospot assay for the diagnosis of active tuberculosis in China.

The rapid diagnosis of smear-negative pulmonary tuberculosis (TB) and extrapulmonary TB is a significant problem in clinical practice. We evaluated the usefulness of a homemade enzyme-linked immunospot (ELISPOT) assay for the diagnosis of active TB in China. Seventy-eight healthy volunteers, 60 patients with active TB, and 32 patients with non-TB diseases were evaluated by tuberculin skin test (TST), an ELISPOT assay using a recombinant CFP-10/ESAT-6 fusion protein (rCFP-10/ESAT-6) as a stimulant, and T-SPOT-TB assay. The spot-forming cells (SFC) from 78 healthy subjects containing both PPD-positive and -negative persons was 3.7 ± 6.5. Among 31 diagnosed TB patients, the ELISPOT assay had a sensitivity of 67.7%, compared to a sensitivity of 77.4% for the T-SPOT-TB assay. The ELISPOT assay was more sensitive in smear-positive TB cases (76.9%) than in smear-negative TB cases (61.1%), while T-SPOT-TB had roughly similar sensitivities in smear-positive (76.9%) and smear-negative TB cases (77.8%). The specificity was 90.6% for ELISPOT and 78.1% for T-SPOT-TB among 32 subjects with non-TB diseases. The SFC of TB cases was significantly higher than that of non-TB disease cases, and the SFC of smear-positive TB cases was significantly higher than that of smear-negative TB cases (P < 0.01). We confirmed that the homemade ELISPOT assay appears more specific for the diagnosis of active TB than T-SPOT-TB. ELISPOT assay may be a useful method for the rapid diagnosis of active TB, especially for cases of smear-negative TB.

Latent tuberculosis infection amongst new recruits to the Chinese army: comparison of ELISPOT assay and tuberculin skin test.

BACKGROUND: In China, latent tuberculosis infection (LTI) is frequent. To protect the health of soldiers and monitor TB infection, new recruits were routinely examined for LTI by tuberculin skin testing (TST). This is the first report on the extent of LTI in the Chinese army comparing TST and enzyme-linked immunospot (ELISPOT) assay. METHODS: New recruits to the army were interviewed, routinely examined and injected intradermally with purified protein derivative (PPD) in March 2007. At the same time, 100 male soldier volunteers were detected with ELISPOT assay using recombinant CFP-10/ESAT-6 fusion protein (rCFP-10/ESAT-6) as a stimulus. RESULTS: The prevalence of LTI, as estimated by TST and ELISPOT assay, was 41% and 21% of new recruits, respectively. Vaccination scars on the arms could be found in 83% of volunteers with positive TST and 19% of volunteers with negative TST. Five individuals with strongly positive TST of whom 2 had negative ELISPOT were not given chemotherapy, and were observed for 20 months. None developed active TB. CONCLUSIONS: The prevalence of LTI in new recruits to the Chinese army is not as high as previously reported. ELISPOT technique may be the most accurate screening method for the TB infection in China, which was not interfered by BCG vaccination.