Longxiang Su

Identification of Novel Biomarkers for Sepsis Prognosis via Urinary Proteomic Analysis Using iTRAQ Labeling and 2D-LC-MS/MS

Objectives Sepsis is the major cause of death for critically ill patients. Recent progress in proteomics permits a thorough characterization of the mechanisms associated with critical illness. The purpose of this study was to screen potential biomarkers for early prognostic assessment of patients with sepsis. Methods For the discovery stage, 30 sepsis patients with different prognoses were selected. Urinary proteins were identified using isobaric tags for relative and absolute quantitation (iTRAQ) coupled with LC-MS/MS. Mass spec instrument analysis were performed with Mascot software and the International Protein Index (IPI); bioinformatic analyses were used by the algorithm of set and the Gene Ontology (GO) Database. For the verification stage, the study involved another 54 sepsis-hospitalized patients, with equal numbers of patients in survivor and non-survivor groups based on 28-day survival. Differentially expressed proteins were verified by Western Blot. Results A total of 232 unique proteins were identified. Proteins that were differentially expressed were further analyzed based on the pathophysiology of sepsis and biomathematics. For sepsis prognosis, five proteins were significantly up-regulated: selenium binding protein-1, heparan sulfate proteoglycan-2, alpha-1-B glycoprotein, haptoglobin, and lipocalin; two proteins were significantly down-regulated: lysosome-associated membrane proteins-1 and dipeptidyl peptidase-4. Based on gene ontology clustering, these proteins were associated with the biological processes of lipid homeostasis, cartilage development, iron ion transport, and certain metabolic processes. Urinary LAMP-1 was down-regulated, consistent with the Western Blot validation. Conclusion This study provides the proteomic analysis of urine to identify prognostic biomarkers of sepsis. The seven identified proteins provide insight into the mechanism of sepsis. Low urinary LAMP-1 levels may be useful for early prognostic assessment of sepsis. Trial Registration ClinicalTrial.gov NCT01493492

Diagnostic value of urine sTREM-1 for sepsis and relevant acute kidney injuries: a prospective study

IntroductionWe explored the diagnostic value of a urine soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) for early sepsis identification, severity and prognosis assessment, and for secondary acute kidney injury (AKI). We compared this with white blood cell (WBC) counts, serum C-reactive protein (CRP), serum procalcitonin (PCT), urine output, creatinine clearance (CCr), serum creatinine (SCr), and blood urea nitrogen (BUN).MethodsWe enrolled 104 subjects admitted to the ICU: 16 cases with systemic inflammatory response syndrome (SIRS); 35 with sepsis and 53 with severe sepsis. Results for urine sTREM-1, WBC, serum CRP and serum PCT were recorded on days 1, 3, 5, 7, 10, and 14. For 17 sepsis cases diagnosed with secondary AKI, comparisons between their urine sTREM-1, urine output, CCr, SCr and BUN at diagnosis and 48 h before diagnosis were made.ResultsOn the day of admission to the ICU, and compared with the SIRS group, the sepsis group exhibited higher levels of urine sTREM-1 and Acute Physiologic Assessment and Chronic Health Evaluation II (APACHE II) scores (P < 0.05). Areas under the curve (AUC) shaped by the scores were 0.797 (95% CI 0.711 to 0.884) and 0.722 (95% CI 0.586 to 0.858), respectively. On days 1, 3, 5, 7, 10, and 14, urine sTREM-1, serum PCT and WBC levels registered higher in the severe sepsis group in contrast to the sepsis group (P < 0.05). Urine sTREM-1 and serum PCT levels continuously increased among non-survivors, while WBC and serum CRP levels in both groups declined. For 17 patients with AKI, urine sTREM-1, SCr and BUN levels at 48 h before AKI diagnosis were higher, and CCr level was lower than those for non-AKI subjects (P < 0.05). AUC for urine sTREM-1 was 0.922 (95% CI 0.850 to 0.995), the sensitivity was 0.941, and the specificity was 0.76 (based on a cut-off point of 69.04 pg/ml). Logistic regression analysis showed that urine sTREM-1 and severity were risk factors related to AKI occurrence.ConclusionsBesides being non-invasive, urine sTREM-1 testing is more sensitive than testing WBC, serum CRP, and serum PCT for the early diagnosis of sepsis, as well as for dynamic assessments of severity and prognosis. It can also provide an early warning of possible secondary AKI in sepsis patients.Trial RegistrationClinicalTrial.gov identifier NCT01333657

Urinary proteomics analysis for sepsis biomarkers with iTRAQ labeling and two-dimensional liquid chromatography-tandem mass spectrometry.

BACKGROUND Proteomics has only recently been applied to the field of critical care research. Sepsis is a major factor contributing to intensive care unit admissions and deaths. The purpose of this study was to screen potential urinary biomarkers for sepsis using A proteomics approach. METHODS Fifteen sepsis and 15 systemic inflammatory response syndrome patients were involved in this study. Urinary proteins were identified by isobaric tag for relative and absolute quantitation coupled with liquid chromatography–tandem mass spectrometry. Mass spectroscopy analysis was performed with the Mascot software and the International Protein Index. Bioinformatics analyses were performed using the hierarchy cluster analysis, the STRING software, the Gene Ontology, and the Kyoto Encyclopedia of Genes and Genome database. RESULTS One hundred thirty proteins were identified, and 34 differentially expressed proteins were selected (fold change, >1.5). On the basis of the Gene Ontology and the Kyoto Encyclopedia of Genes and Genome database, these 34 proteins were identified to be involved in inflammation, immunity, and structural or cytoskeletal processes. Five proteins were selected by a protein–protein interaction network for sepsis differentiation: cadherin 1, haptoglobin, complement 3, alpha-1-antitrypsin, and ceruloplasmin. CONCLUSION Urinary proteomics may represent a suitable approach for sepsis-related research. The detection of urinary biomarkers is expected to become a noninvasive and acceptable method, which facilitates the close surveillance of diseases and reduces medical costs. LEVELS OF EVIDENCE Diagnostic study, level IV.

Dynamic Changes in Serum Soluble Triggering Receptor Expressed on Myeloid Cells-1 (sTREM-1) and its Gene Polymorphisms are Associated with Sepsis Prognosis

This study explored the association of sepsis prognosis with dynamic changes in serum soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) and its polymorphisms. We enrolled 80 subjects with sepsis and 80 controls. Serum sTREM-1 was tested on days 1, 3, 5, 7, 10, and 14. PCR sequencing was performed to detect TREM-1 genetic variation on its four exons. sTREM-1 levels were significantly higher in the nonsurvivor than in the survivor group (pu2009<u20090.001), and those at each time point were the same (pu2009≤u20090.001). Of the three tested TREM-1 SNPs (rs144672509, rs2234237, and rs2234246), only rs2234237 (Ser25Thr) was significantly associated with sepsis prognosis in three inheritance models (pu2009<u20090.05). However, there was no relationship between TREM-1 polymorphism and dynamic concentration of sTREM-1. Logistic regression showed that sTREM-1, APACHE II, and rs2234237 polymorphism are risk factors for prognosis. Dynamic changes in serum sTREM-1 and rs2234237 polymorphism could be used in sepsis prognosis assessment.

Genomic evolution of 11 type strains within family Planctomycetaceae.

The species in family Planctomycetaceae are ideal groups for investigating the origin of eukaryotes. Their cells are divided by a lipidic intracytoplasmic membrane and they share a number of eukaryote-like molecular characteristics. However, their genomic structures, potential abilities, and evolutionary status are still unknown. In this study, we searched for common protein families and a core genome/pan genome based on 11 sequenced species in family Planctomycetaceae. Then, we constructed phylogenetic tree based on their 832 common protein families. We also annotated the 11 genomes using the Clusters of Orthologous Groups database. Moreover, we predicted and reconstructed their core/pan metabolic pathways using the KEGG (Kyoto Encyclopedia of Genes and Genomes) orthology system. Subsequently, we identified genomic islands (GIs) and structural variations (SVs) among the five complete genomes and we specifically investigated the integration of two Planctomycetaceae plasmids in all 11 genomes. The results indicate that Planctomycetaceae species share diverse genomic variations and unique genomic characteristics, as well as have huge potential for human applications.

The synergistic therapeutic effect of hepatocyte growth factor and granulocyte colony-stimulating factor on pulmonary hypertension in rats.

Pulmonary arterial hypertension (PAH) is characterized by a progressive increase in pulmonary arterial pressure and vascular resistance. Despite advances in therapy for PAH, its treatment and prognosis remain poor. We aimed to investigate whether the transplantation of bone marrow mesenchymal stem cells (MSCs) overexpressing hepatocyte growth factor (HGF), alone or in combination with granulocyte colony-stimulating factor (G-CSF), attenuates the development of experimental monocrotaline (MCT)-induced PAH. Three weeks after MCT administration, rats were divided into the following groups: (1) untreated (PAH); (2) HGF treated; (3) MSCs administered; (4) HGF-MSCs treated; and (5) HGF-MSCs plus G-CSF treated. After 3 weeks, hemodynamic changes, histomorphology, and angiogenesis were evaluated. To elucidate the molecular mechanisms of vascular remodeling and angiogenesis, serum levels of transforming growth factor (TGF)-β and endothelin-1 (ET-1) were measured, and the gene and protein expression levels of vascular cell adhesion molecule-1 (VCAM-1) and matrix metalloproteinase-9 (MMP-9) were determined. Compared with the PAH, MSC, and G-CSF groups, the HGF and HGF+G-CSF groups exhibited significantly reduced right ventricular hypertrophy and mean pulmonary arterial pressure (P < 0.05). Histologically, vessel muscularization or thickening and collagen deposition were also significantly decreased (P < 0.05). The number of vessels in the HGF+G-CSF group was higher than that in the other groups (P < 0.05). The TGF-β and ET-1 concentrations in the plasma of pulmonary hypertensive rats were markedly lower in the HGF and HGF+G-CSF groups (P < 0.05). Furthermore, HGF induced the expression of VCAM-1, and HGF treatment together with G-CSF synergistically stimulated MMP-9 expression. Transplanted HGF-MSCs combined with G-CSF potentially offer synergistic therapeutic benefit for the treatment of PAH.

Diagnostic value of urine sCD163 levels for sepsis and relevant acute kidney injury: a prospective study

BackgroundSepsis is a common syndrome in critically ill patients and easily leads to the occurrence of acute kidney injury (AKI), with high mortality rates. This study aimed to investigate the diagnostic value of urine soluble CD163 (sCD163) for identification of sepsis, severity of sepsis, and for secondary AKI, and to assess the patients’ prognosis.MethodsWe enrolled 20 cases with systemic inflammatory response syndrome (SIRS), 40 cases with sepsis (further divided into 17 sepsis cases and 23 severe sepsis cases) admitted to the intensive care unit (ICU), and 20 control cases. Results for urine sCD163 were recorded on the day of admission to the ICU, and AKI occurrence was noted.ResultsOn the day of ICU admission, the sepsis group exhibited higher levels of urine sCD163 (74.8 ng/ml; range: 47.9-148.3 ng/ml) compared with those in the SIRS group (31.9 ng/ml; 16.8-48.0, Pu2009<u20090.001). The area under the curve (AUC) was 0.83 (95% confidence interval [CI]: 0.72-0.94, Pu2009<u20090.001) the sensitivity was 0.83, and the specificity was 0.75 (based on a cut-off point of 43.0 ng/ml). Moreover, the severe sepsis group appeared to have a higher level of sCD163 compared with that in the sepsis group (76.2; 47.2-167.5 ng/ml vs. 74.2; 46.2-131.6 ng/ml), but this was not significant. For 15 patients with AKI, urine sCD163 levels at AKI diagnosis were significantly higher than those of the remaining 35 sepsis patients upon ICU admission (121.0; 74.6-299.1 ng/ml vs. 61.8; 42.8-128.3 ng/ml, Pu2009=u20090.049). The AUC for urine sCD163 was 0.688 (95% CI: 0.51-0.87, Pu2009=u20090.049). Sepsis patients with a poor prognosis showed a higher urine sCD163 level at ICU admission (98.6; 50.3-275.6 ng/ml vs. 68.0; 44.8-114.5 ng/ml), but this was not significant. Patients with AKI with a poor prognosis had higher sCD163 levels than those in patients with a better prognosis (205.9; 38.6-766.0 ng/ml vs. 80.9; 74.9-141.0 ng/ml), but this was not significant.ConclusionsThis study shows, for the first time, the potential value of urine sCD163 levels for identifying sepsis and diagnosing AKI, as well as for assessment of patients’ prognosis.Trial RegistrationChiCTR-ONC-10000812

Effects of Vitamin B6 Therapy for Sepsis Patients with Linezolid-Associated Cytopenias: A Retrospective Study

Background The common adverse effects of linezolid for treating septic patients with gram-positive cocci is anemia and thrombocytopenia, which limit its clinical application. Objectives We determined the effects of vitamin B6 adjunctive therapy on linezolid-associated cytopenias, and retrospectively studied 75 septic patients who received at least 7 days of linezolid treatment. Methods Patients were divided into a linezolid treatment group (LTG; n = 41) that received linezolid only and a combination treatment group (CTG; n = 34) that received both linezolid and vitamin B6. Each group was further subdivided into those with sepsis and those with severe sepsis. Each patient had red blood cell (RBC), hemoglobin (Hb), hematocrit (Hct), and platelet (PLT) measurements at baseline (day 0) and every other day for 2 weeks during treatment; these parameters were compared between the groups and assessed for time-dependent trends. Results For patients in the LTG, RBC, Hb, and Hct values showed statistically significant reductions over time, and these values were lower compared with the values in the CTG. The CTG also showed downward trends, except on the first day of treatment. The PLT count also decreased in both groups. Patients with severe sepsis had lower PLT counts in both treatment groups compared with the septic patients. Conclusions Septic patients who received a combination treatment of linezolid and vitamin B6 might show positive effects for linezolid-associated reductions in some hematologic parameters (RBC, Hb, and Hct). This combined treatment might also slow PLT reduction, which was more evident in patients with severe sepsis. ClinicalTrials.gov identifier: NCT01295801.

Genome Sequence of Bacillus cereus Strain LCT-BC235, Carried by the Shenzhou VIII Spacecraft

ABSTRACT In order to explore the effect of space environments on Bacillus cereus, we determined the draft genome sequence of a B. cereus strain, LCT-BC235, which was isolated after space flight.

The efficacy of MSC-HGF in treating pulmonary arterial hypertension (PAH) and connexin remodelling

BackgroundThis study investigated whether the hepatocyte growth factor (HGF) genetically modified marrow-mesenchymal stem cells (MSCs) transplantation could offer a therapeutic benefit for pulmonary arterial hypertension (PAH).MethodologyThree weeks after monocrotaline administration, Sprague-Dawley rats were randomly divided into the following groups: PAH (n=10), MSCs (5×106 MSCs injected into the jugular veins, n=10), HGF (5×106 MSCs transfected with Ad-HGF into the jugular veins, n=10). Another three weeks later, hemodynamic changes and histomorphology were observed. Electron microscopy and immunofluorescence were also used to observe changes in the gap junctions of the heart.ResultsCompared with the PAH and MSC groups, hemodynamic parameters improved significantly in the MSC-HGF group. Right ventricular hypertrophy was improved as measured by the RV/LV weight and thickness ratios. Histologically, cardiac myocytes and cell nuclei recovered and interstitial fibrosis decreased in the MSC and MSC-HGF groups. Under electron microscopy, the gap junctions exhibited a disorganised morphology in the PAH group and the number of gap junctions was lower in this group than in the other groups. The distribution of connexins 43 and 40 were improved in the MSC-HGF group.ConclusionsMCT-induced PAH can be treated and improved by HGF genetically modified MSCs, which may occur via connexin remodeling.