Plasma Metabolic Profile Determination in Young ST-segment Elevation Myocardial Infarction Patients with Ischemia and Reperfusion: Ultra-performance Liquid Chromatography and Mass Spectrometry for Pathway Analysis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>This study was to establish a disease differentiation model for ST-segment elevation myocardial infarction (STEMI) youth patients experiencing ischemia and reperfusion via ultra-performance liquid chromatography and mass spectrometry (UPLC/MS) platform, which searches for closely related characteristic metabolites and metabolic pathways to evaluate their predictive value in the prognosis after discharge.</p><p><b>METHODS</b>Forty-seven consecutive STEMI patients (23 patients under 45 years of age, referred to here as "youth," and 24 "elderly" patients) and 48 healthy control group members (24 youth, 24 elderly) were registered prospectively. The youth patients were required to provide a second blood draw during a follow-up visit one year after morbidity (n = 22, one lost). Characteristic metabolites and relative metabolic pathways were screened via UPLC/MS platform base on the Kyoto encyclopedia of genes and genomes (KEGG) and Human Metabolome Database. Receiver operating characteristic (ROC) curves were drawn to evaluate the predictive value of characteristic metabolites in the prognosis after discharge.</p><p><b>RESULTS</b>We successfully established an orthogonal partial least squares discriminated analysis model (R2X = 71.2%, R2Y = 79.6%, and Q2 = 55.9%) and screened out 24 ions; the sphingolipid metabolism pathway showed the most drastic change. The ROC curve analysis showed that ceramide [Cer(d18:0/16:0), Cer(t18:0/12:0)] and sphinganine in the sphingolipid pathway have high sensitivity and specificity on the prognosis related to major adverse cardiovascular events after youth patients were discharged. The area under curve (AUC) was 0.671, 0.750, and 0.711, respectively. A follow-up validation one year after morbidity showed corresponding AUC of 0.778, 0.833, and 0.806.</p><p><b>CONCLUSIONS</b>By analyzing the plasma metabolism of myocardial infarction patients, we successfully established a model that can distinguish two different factors simultaneously: pathological conditions and age. Sphingolipid metabolism is the top most altered pathway in young STEMI patients and as such may represent a valuable prognostic factor and potential therapeutic target.</p>

Analysis of serum metabolic profile by ultra-performance liquid chromatography-mass spectrometry for biomarkers discovery: application in a pilot study to discriminate patients with tuberculosis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Tuberculosis (TB) is a chronic wasting inflammatory disease characterized by multisystem involvement, which can cause metabolic derangements in afflicted patients. Metabolic signatures have been exploited in the study of several diseases. However, the serum that is successfully used in TB diagnosis on the basis of metabolic profiling is not by much.</p><p><b>METHODS</b>Orthogonal partial least-squares discriminant analysis was capable of distinguishing TB patients from both healthy subjects and patients with conditions other than TB. Therefore, TB-specific metabolic profiling was established. Clusters of potential biomarkers for differentiating TB active from non-TB diseases were identified using Mann-Whitney U-test. Multiple logistic regression analysis of metabolites was calculated to determine the suitable biomarker group that allows the efficient differentiation of patients with TB active from the control subjects.</p><p><b>RESULTS</b>From among 271 participants, 12 metabolites were found to contribute to the distinction between the TB active group and the control groups. These metabolites were mainly involved in the metabolic pathways of the following three biomolecules: Fatty acids, amino acids, and lipids. The receiver operating characteristic curves of 3D, 7D, and 11D-phytanic acid, behenic acid, and threoninyl-γ-glutamate exhibited excellent efficiency with area under the curve (AUC) values of 0.904 (95% confidence interval [CI]: 0863-0.944), 0.93 (95% CI: 0.893-0.966), and 0.964 (95% CI: 00.941-0.988), respectively. The largest and smallest resulting AUCs were 0.964 and 0.720, indicating that these biomarkers may be involved in the disease mechanisms. The combination of lysophosphatidylcholine (18:0), behenic acid, threoninyl-γ-glutamate, and presqualene diphosphate was used to represent the most suitable biomarker group for the differentiation of patients with TB active from the control subjects, with an AUC value of 0.991.</p><p><b>CONCLUSION</b>The metabolic analysis results identified new serum biomarkers that can distinguish TB from non-TB diseases. The metabolomics-based analysis provides specific insights into the biology of TB and may offer new avenues for TB diagnosis.</p>

Examination of anti-HBx in sera from patients of chronic hepatitis B,liver cirrhosis and hepatocellular carcinoma and its clinical significance / 中华检验医学杂志

Objective To establish a method of detecting hepatitis B virus x antigen (HBxAg) and antibody to HBxAg (anti-HBx) and to demonstrate its clinical significance of HBxAg and anti-HBx in sera from patients of chronic hepatitis B (CHB),liver cirrhosis (LC) and hepatocellular carcinoma (HCC). Methods Full length HBx gene was cloned into pET30a(+),a prokaryotic expression vector,named pET30a-X.It was transformed into Escherichia coli BL21 (DE3),followed the fusion protein of HBx-His was induced by IPTG.The purified fusion protein was used to immunize rabbit as an antigen to generate polyclonal antibody to HBx protein.The method of enzyme-linked immunosorbent assay (ELISA) was established by using purified fusion protein and generated antibody,which was used to detect HBxAg and anti-HBx in sera from patients of CHB,LC,HCC and normal healthy people.Results The positive rates of HBxAg/anti-HBx were 8.7%/10.4% for CHB,17.9%/40.6% for LC,and 9.8%/34.4% for HCC, respectively.In statistics,the positive rates of anti-HBx in LC and HCC were higher than that in CHB (P