Identification of Key Immune-Related Genes in the Progression of Septic Shock.

Objective: Septic shock is the severe complication of sepsis, with a high mortality. The inflammatory response regulates the immune status and mediates the progression of septic shock. In this study, we aim to identify the key immune-related genes (IRGs) of septic shock and explore their potential mechanism. Methods: Gene expression profiles of septic shock blood samples and normal whole blood samples were retrieved from the Gene Expression Omnibus (GEO) and Genotype-Tissue Expression Portal (GTEx). The differential expression genes (DEGs) and septic shock-specific immune-related genes (SSSIRGs) were evaluated and identified, along with the immune components by "cell type identification by estimating relative subsets of RNA transcripts (CIBERSORT, version x)" algorithm. Additionally, in order to explore the key regulatory network, the relationship among SSSIRGs, upstream transcription factors (TFs), and downstream signaling pathways were also identified by Gene Set Variation Analysis (GSVA) and co-expression analysis. Moreover, the Connectivity Map (CMap) analysis was applied to find bioactive small molecules against the members of regulation network while Chromatin Immunoprecipitation sequencing (ChIP-seq) and Assay for Targeting Accessible-Chromatin with high-throughput sequencing (ATAC-seq) data were used to validate the regulation mechanism of the network. Results: A total of 14,843 DEGs were found between 63 septic shock blood samples and 337 normal whole blood samples. Then, we identified septic shock-specific 839 IRGs as the intersection of DEGs and IRGs. Moreover, we uncovered the regulatory networks based on co-expression analysis and found 28 co-expression interaction pairs. In the regulation network, protein phosphatase 3, catalytic subunit, alpha isozyme (PPP3CA) may regulate late estrogen response, glycolysis and TNFα signaling via NFκB and HLA; Kirsten rat sarcoma viral oncogene homolog (KRAS) may be related to late estrogen response and HLA; and Toll-like receptor 8 (TLR8) may be associated with TNFα signaling via NFκB. And the regulation mechanisms between TFs and IRGs (TLR8, PPP3CA, and KRAS) were validated by ChIP-seq and ATAC-seq. Conclusion: Our data identify three SSSIRGs (TLR8, PPP3CA, and KRAS) as candidate therapeutic targets for septic shock and provide constructed regulatory networks in septic shock to explore its potential mechanism.

[Risk factors and their predictive value for intensive care unit acquired weakness in patients with sepsis].

OBJECTIVE: To explore the risk factors of intensive care unit acquired weakness (ICUAW) in patients with sepsis, and to evaluate the predictive value of each risk factor for ICUAW. METHODS: A case control study was conducted, 60 septic patients admitted to the intensive care unit (ICU) of Henan Provincial People's Hospital from October 20, 2020 to February 20, 2021 were enrolled. The patients were divided into two groups: sepsis ICUAW group and sepsis non-ICUAW group. The data of gender, age, body mass index (BMI), acute physiology and chronic health evaluation II (APACHE II) score, complications, mechanical ventilation, duration of ICUAW, length of stay in ICU, fasting blood glucose, blood lactic acid (Lac), procalcitonin (PCT), C-reactive protein (CRP), sequential organ failure assessment (SOFA) score, outcome, antimicrobial agent, glucocorticoid, sedatives and analgesics drugs and vasoactive drugs were collected. Risk factors were screened by univariate Logistic regression analysis, and odds ratio (OR) was adjusted by multivariate binary logistic regression, P < 0.05 was considered as independent risk factors. Finally, the receiver operating characteristic curve (ROC curve) was drawn to analyze the predictive value of independent risk factors. RESULTS: The APACHE II score of the sepsis ICUAW group was significantly higher than that of the sepsis non-ICUAW group (23.05±8.17 vs. 15.33±4.89, P < 0.05), the total length of stay in the ICU was significantly longer than that of the sepsis non-ICUAW group (days: 15.1±9.2 vs. 8.5±3.4, P < 0.05), the improvement rate of patients was significantly lower than that of the sepsis non-ICUAW group [45.0% (9/20) vs. 95.0% (38/40), P < 0.05]. After univariate Logistic regression and multicollinearity test analysis, 7 factors including APACHE II score, average SOFA score, blood lactic acid, proportion of mechanical ventilation, sedatives and analgesics drugs, type of antibiotics and type of vasoactive drugs were included in the binary Logistic regression model [OR: 1.21, 2.05, 2.26, 0.21, 1.54, 2.07, 1.38, 95% confidence interval (95%CI): 1.09-1.35, 1.42-2.94, 1.12-4.57, 0.05-0.66, 1.03-2.29, 1.27-3.37, 0.96-2.00, all P < 0.05]. Hosmer-Lemchaw test P = 0.901, and the correct percentage of prediction was 85%, indicating good model fit. Multivariate binary Logistic regression analysis showed that APACHE II score and average SOFA score were independent risk factors for the occurrence of ICUAW in septic patients (APACHE II score: OR = 1.17, 95%CI was 1.004-1.376, P = 0.044; average SOFA score: OR = 1.86, 95%CI was 1.157-2.981, P = 0.01). ROC curve analysis showed that the mean value of APACHE II score, average SOFA score and their combined detection had a certain predictive value for the occurrence of ICUAW in sepsis patients, areas under ROC curve (AUC) were 0.787, 0.881, 0.905, 95%CI was 0.646-0.928, 0.791-0.972, 0.828-0.982, all P < 0.05. When the cut-off value was 19.500, 6.225, 0.375, the sensitivity was 75%, 90%, 90%, and the specificity were 80%, 80%, 85%, respectively. CONCLUSIONS: APACHE II score and average SOFA score can be used as independent risk factors for the occurrence of ICUAW in sepsis, and their combined predictive value is better than that of individual index.

Amine-Aldehyde Chemical Conjugation on a Potassium Hydroxide-Treated Polystyrene ELISA Surface for Nanosensing an HIV-p24 Antigen.

The enzyme-linked immunosorbent assay (ELISA) has been widely used for disease surveillance and drug screening due to its relatively higher accuracy and sensitivity. Fine-tuning the ELISA is mandatory to elevate the specific detection of biomolecules at a lower abundance. Towards this end, higher molecular capture on the polystyrene (PS) ELISA surface is crucial for efficient detection, and it could be attained by immobilizing the molecules in the correct orientation. It is highly challenging to immobilize protein molecules in a well-aligned manner on an ELISA surface due to charge variations. We employed a 3-(aminopropyl) triethoxysilane (APTES)- and glutaraldehyde (GLU)-coupled PS surface chemical strategy to demonstrate the high performance with ELISA. A potassium hydroxide treatment followed by an equal ratio of 1% APTES and GLU attachment was found to be optimal, and a longer incubation with GLU favored maximum sensitivity. p24 is a vital early secreting antigen for diagnosing human immunodeficiency virus (HIV), and it has been used for efficient detection with the above chemistry. Three different procedures were followed, and they led to the improved detection of the HIV-p24 antigen at 1 nM, which is a 30-fold higher level compared to a conventional ELISA surface. The surface chemical functionalization shown here also displays a higher specificity with human serum and HIV-TAT. The above approach with the designed surface chemistry could also be recommended for disease diagnosis on other sensing surfaces involving the interaction of the probe and the analyte in heterogeneous test samples.

[Early assessment value of brain function prognosis in patients with traumatic brain injury by regional saturation of cerebral oxygenation combined with percentage of α variability].

OBJECTIVE: To explore the usability of regional saturation of cerebral oxygenation (rScO2) combined with percentage of α variability (PAV) in predicting brain function prognosis in patients with traumatic brain injury (TBI). METHODS: A retrospective analysis was conducted. The clinical data of patients with TBI who were monitored rScO2 and bedside quantitative electroencephalogram (qEEG) admitted to intensive care unit (ICU) of Henan Provincial People's Hospital from August 2018 to July 2019 were collected. The rScO2, PAV, and Glasgow coma scale (GCS) score were recorded within 72 hours after the TBI. The primary prognostic indicator was the 3-month Glasgow outcome score (GOS) score. The differences between the two groups of poor prognosis of brain function (GOS score 1-3) and good prognosis (GOS score 4-5) were compared. Binary multivariate Logistic regression analysis was used to analyze the correlation between rScO2, PAV, GCS score and the prognosis of brain function in patients with TBI. In addition, receiver operating characteristic (ROC) curve was plotted to analyze the predicting value of rScO2 and PAV only or combination for prognosis of brain function. RESULTS: A total of 42 patients with TBI were enrolled in the study, with rScO2 ≥ 0.60 (grade I) in 14 patients, 0.50 ≤ rScO2 < 0.60 (grade II) in 16 patients, and rScO2 < 0.50 (grade III) in 12 patients. PAV 3-4 scores (grade I) were detected in 16 patients, 2 scores (grade II) in 17 patients, and 1 score (grade III) in 9 patients. GCS score 9-14 (grade I) were observed in 13 patients, 4-8 (grade II) in 23 patients, and 3 (grade III) in 6 patients; 18 patients had poor prognosis and 24 had good one. The rScO2, PAV and GCS scores of the poor-prognosis group were significantly higher than those in the good-prognosis group [rScO2 with grade III: 55.6% (10/18) vs. 8.3% (2/24), PAV with grade III: 38.9% (7/18) vs. 8.4% (2/24), GCS score with grade III: 27.7% (5/18) vs. 4.1% (1/24)] with significant differences (all P < 0.05). There was no significant difference in other general data including gender, age, total length of hospital stay or acute physiology and chronic health evaluation II (APACHE II) score between the two groups. Binary multivariate Logistic regression analysis showed that rScO2 and PAV were independent risk factors for prognosis of brain in patients with TBI [rScO2: odds ratio (OR) = 4.656, 95% confidence interval (95%CI) was 1.071-20.233, P = 0.040; PAV: OR = 3.525, 95%CI was 1.044-11.906, P = 0.042]. ROC curve analysis showed that both of rScO2 and PAV had predictive value for the prognosis of brain function in patients with TBI (AUC was 0.796 and 0.780, respectively, both P < 0.01), and rScO2 combined with PAV had higher predictive value with the AUC of 0.851 (P < 0.01) than rScO2 or PAV alone, the sensitivity was 94.4% and the specificity was 62.5%. CONCLUSIONS: rScO2 and PAV were associated with early brain function prognosis in patients with TBI. The combination of two monitoring indicators can reliably assess the prognosis of brain function in patients with TBI.

[Influence of simvastatin treatment on Toll-like receptor 4 in monocytes of peripheral blood in patients with sepsis and severe sepsis].

OBJECTIVE: To investigate the influence of simvastatin treatment on Toll-like receptor 4 (TLR4) in monocytes of peripheral blood in patients with sepsis and severe sepsis and its significance. METHODS: A prospective randomized controlled trial was conducted. 106 patients with sepsis and 92 patients with severe sepsis admitted to Department of Critical Care Medicine of Henan Provincial People's Hospital from August 2013 to June 2015 were enrolled. These two groups of patients were randomized into conventional treatment group and simvastatin group. All patients received treatment according to the 2012 International Sepsis Treatment Guidelines, including anti-infection drugs, nutritional support, and palliative treatment, and the patients with severe sepsis were given early goal-directed therapy (EGDT). The patients in simvastatin group received simvastatin 40 mg daily orally for at least 15 days. The peripheral blood was collected and the monocytes were isolated at 1, 5, 10, 15 days after intensive care unit (ICU) admission. TLR4 expression on the surface of TLR4/CD14(+) double positive monocytes was determined by flow cytometry, and adverse reaction was observed during treatment. RESULTS: TLR4 expression on the surface of monocytes showed a tendency of decreasing with prolongation of simvastatin treatment in the simvastatin group in patients with sepsis (n = 59) or severe sepsis (n = 54). However, in patients with sepsis, TLR4 level was significantly decreased from 10 days in simvastatin group as compared with that of conventional therapy group (n = 47), and it was decreased up to 15 days [mean fluorescence intensity (MFI): 21 (19, 28) vs. 27 (25, 33) at 10 days, Z = 2.198, P = 0.021; 16 (15, 21) vs. 26 (23, 34) at 15 days, Z = 4.611, P = 0.002]. In patients with severe sepsis, there was no significant difference in TLR4 level at different time points between simvastatin group and conventional treatment group (n = 38) [MFI: 55 (52, 63) vs. 56 (48, 65) at 1 day, Z = 0.313, P = 0.692; 47 (42, 56) vs. 49 (41, 58) at 5 days, Z = 0.827, P = 0.533; 40 (35, 42) vs. 42 (37, 45) at 10 days, Z = 1.012, P = 0.301; 33 (30, 38) vs. 38 (35, 41) at 15 days, Z = 0.539, P = 0.571]. No adverse reaction related with simvastatin was found during treatment in patients with sepsis or severe sepsis. CONCLUSIONS: Statins could significantly down-regulate the TLR4 expression on peripheral blood monocytes in septic patients, while it showed no significant influence on TLR4 expression in patients with severe sepsis. A different effect of statins on TLR4 expression and the downstream inflammation process in sepsis and severe sepsis patients might partially explain the discrepancy in previous reports about the therapeutic effect of statins therapy in sepsis and severe sepsis patients.

Different regulation of Toll-like receptor 4 expression on blood CD14(+) monocytes by simvastatin in patients with sepsis and severe sepsis.

We have demonstrated that regulation of Toll-like receptor 4 (TLR4) surface expression levels on blood CD14(+) monocytes by simvastatin treatment in patient with sepsis is different from that in patients with severe sepsis. In patients with sepsis simvastatin treatment statistically significantly decreased TLR4 surface expression level on blood CD14(+) monocytes, while in patients with severe sepsis simvastatin treatment had no significant influence on TLR4 surface expression level on blood CD14(+) monocytes. The changes of plasma interleukin-6 (IL-6) induced by simvastatin in patients with sepsis and severe sepsis were similar with that of TLR4. Our results indicated simvastatin treatment differently influenced inflammation process in patients with sepsis and severe sepsis, which might partially explain the discrepancy, presented by previous trials, about the therapeutic effects of simvastatin treatment in patients with sepsis and severe sepsis.

[Characteristics and treatment of multiple organ dysfunction syndrome in patients with trauma in the Wenchuan earthquake on 12th May, 2008].

OBJECTIVE: To analyze the characteristics and treatment of the multiple organ dysfunction syndrome (MODS) in patients in the Wenchuan earthquake on 12th May, 2008, in order to provide theoretical reference for future care for such patients. METHODS: Characteristics of MODS in these patients were analyzed, differences between survivors and non-survivors were compared, and therapeutic measures, and the time of the treatment for MODS in patients with earthquake related injury or illness who were admitted to West China Hospital from 12th May to 20th June, 2008, were retrospectively analyzed. RESULTS: A total of 42 MODS patients were admitted to intensive care unit (ICU). Both the acute physiology and chronic health evaluation II (APACHE II) score and predicted death risk were lowering during the course of therapy. Fractures of bones of extremities were predominant in the earthquake related diseases, with an incidence of 45.2%. The actual mortality of MODS (33.3%) was lower than the predicted death risk (41.5%). The age, the time of receiving the first treatment in ICU after the earthquake, the Glasgow score, the oxygen index, blood creatinine level, platelet count, and vasoactive agent pumping velocity were significantly different between survivors and non-survivors (all P<0.05). The overall mortality was 9.8%, the morbidity of cardiac dysfunction, the incidence of acute renal failure (ARF) and sepsis were significantly different between non-survivors and survivors (all P<0.05). The use of mechanical ventilation, continuous renal replacement therapy (CRRT), and vasoactive agent reached peak level on the 14-29 days after the earthquake. CONCLUSION: Fracture of bones of extremities are predominant injury in the earthquake related diseases, and the cause of death is closely associated with multiple trauma and ARF, systemic infection of large wound surfaces. The central nervous system, respiratory system, circulatory system, renal function, circulatory system should be monitored during the treatment. Adequate preparedness is essential in order to cope with the peak period of occurrence of serious complications after a disaster.