Advances in pathophysiology, diagnosis and treatment of adult severe-associated thrombotic microangiopathy / 中华危重病急救医学

Thrombotic microangiopathy (TMA) is a group of highly heterogeneous, acute and severe clinicopathological syndromes, characterized by microangiopathic hemolytic anemia (MAHA), thrombocytopenia and ischemic injury of end organs. TMA has the characteristics of dangerous condition, multiple organ involvement and high mortality. Patients with severe TMA need to be admitted to intensive care unit (ICU) for organ function support therapy. Early and rapid evaluation, differential diagnosis, and timely and effective treatment are the key to improve the prognosis of TMA patients. Here, we review the pathophysiological changes, diagnosis differential diagnosis, and treatment of the severe TMA in adult.

Differential gene expression and bioinformatics analysis in sepsis secondary to pneumonia / 中华危重病急救医学

Objective:To analyze and screen the key genes of sepsis secondary to pulmonary infection by bioinformatics, and to provide theoretical basis for the basic research of the disease and find an ideal animal model program.Methods:Experiment 1 (bioinformatics analysis): gene expression data sets of pulmonary infection secondary sepsis patients and multiple sepsis animal models were screened by Gene Expression Omnibus (GEO) Database, and gene differences were analyzed by R software. Differential genes were analyzed by gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Correlation analysis was conducted between differential genes and clinical symptoms in the data set of pulmonary infection secondary sepsis, and the correlation heat map between differential genes and clinical symptoms was drawn. Key genes were screened by weighted gene co-expression network analysis (WGCNA) and protein-protein interaction network analysis (PPIN) clustering. Experiment 2 (sepsis animal model preparation): male mice weighing 21-25 g were randomly divided into the key genes group and the control (Sham) group. And cecal ligation and puncture (CLP) was used to establish mouse sepsis model, while the mice in sham group were performed by exposure of cecum. And all the mice were scarified 24 hours after surgery to extract the total RNA from lung tissue, real time fluorescent quantitative polymerase chain reaction (RT-qPCR) was used to detect mRNA expression of key genes.Results:Experiment 1 (bioinformatics analysis): 319 differential genes were showed by GSE 134364 and GSE 65682 data set analysis of pulmonary infection secondary sepsis. And there was no genetic difference between community acquired pneumonia (CAP) and hospital acquired pneumonia (HAP) in patients with pulmonary infection secondary to sepsis. Obvious differences existed between differential genes in animal models, and there was no common differential gene. Differential genes in patients and animal models were similarly enriched in GO function, mainly in cell differentiation, regulation of cell process, and regulation of cellular response to stimuli, there were significant differences in pathway enrichment, among which, CLP animal models showed higher consistency with patients. The key genes obtained by WGCNA and PPIN analysis were MAPK14, NLRC4 and LCN2. Experiment 2 (sepsis animal model preparation): animal experiment results showed that the mRNA expressions of MAPK14, NLRC4 and LCN2 in lung tissue of CLP model mice were significantly up-regulated compared with the sham group.Conclusions:MAPK14, NLRC4 and LCN2 are key genes involved in the regulation of biological processes of pulmonary sepsis secondary to infection, and are potential research directions of this disease. What's more, CLP animal model can better reflect the biological characteristics of patients with pulmonary infection secondary sepsis, and is one of the ideal animal model schemes for pulmonary infection secondary sepsis.

Analysis of clinical characteristics of patients with different vaccines and underlying diseases infected with novel coronavirus Omicron variant / 中华危重病急救医学

Objective:To analyze the clinical characteristics of patients inoculated with different vaccines and underlying diseases, infected with the novel coronavirus Omicron variant.Methods:The data of 430 patients infected with the novel coronavirus Omicron variant who were admitted to Tianjin First Center Hospital from January 21, 2022 to March 7, 2022 were collected. A total of 108 patients with Omicron variant infection with underlying diseases were selected and enrolled. The gender, age, body mass index (BMI), history of underlying diseases, vaccination status (vaccination times, vaccination type), clinical symptoms, laboratory test indicators, imaging data, hospitalization time, nucleic acid negative conversion time, re-positivity and antibody titer from the two groups of the patients were collected and analyzed.Results:In the 108 patients, 93 cases received inactivated vaccine and 15 cases received adenovirus vaccine. There was no statistically significant difference between the two groups in terms of gender, age, BMI, disease types, whether completed the fully vaccinated, whether had prime boost and underlying diseases. Both groups had fever, dry cough, sore throat, runny nose and other clinical symptoms, but there were no statistical difference between the two groups. There were no statistically significant differences in laboratory blood routine tests, biochemical indexes, C-reactive protein (CRP) level and the results of chest computed tomography (CT) imaging between the two groups. There were no statistically significant differences in hospitalization days, nucleic acid negative conversion time, whether admission to intensive care unit (ICU), turn re-positive on nucleic acid tests and immunoglobulin M (IgM) antibody titer expression between the two groups, but immunoglobulin G (IgG) antibody titer in adenovirus group was higher than that in inactivated group (g/L: 229.67±26.13 vs. 194.33±61.56, P = 0.020). There were also no significant differences in laboratory examinations, hospitalization days, nucleic acid negative conversion time, turn re-positive on nucleic acid tests and Novel coronavirus antibody titers expression of the patients with booster shots between the inactivated vaccine group and the adenovirus vaccine group. Conclusions:The protection of inactivated virus vaccine is equivalent to adenovirus vaccine in patients with underlying disease Omicron variant infection, and the titer of IgG antibody in patients with adenovirus vaccine is higher than that in patients with inactivated virus vaccine after one week of recovery.

Effect of hypophosphatemia on the prognosis of critically ill patients / 中华危重病急救医学

Objective:To explore the impact of hypophosphatemia on the occurrence and prognosis of critically ill patient.Methods:The clinical data of critically ill patients admitted to the intensive care unit (ICU) of Tianjin First Central Hospital from October 2021 to April 2022 were retrospectively analyzed. Patients were divided into hypophosphatemia group (serum phosphorus level < 0.80 mmol/L) and non-hypophosphatemia group (serum phosphorus level ≥ 0.80 mmol/L) when they were admitted to the ICU. The following variables were also collected, including gender, age, acute physiology and chronic health evaluationⅡ(APACHE Ⅱ), sequential organ failure assessment (SOFA), serum phosphorus level, serum calcium level, serum magnesium level, white blood cell count (WBC), neutrophil count (NEU), lymphocyte count (LYM), neutrophil/lymphocyte ratio (NLR), C-reactive protein (CRP), presence of infection and infection site, length of hospital stay, ICU stay, 28-day mortality, and mechanical ventilation time. Multivariate Logistic regression analysis was used to evaluate the relationship between each variable and the 28-day mortality. The receiver operator characteristic curve (ROC curve) was drawn, and the area under the ROC curve (AUC) and 95% confidence interval (95% CI) were calculated to evaluate the predictive value of serum phosphorus levels for the prognosis of ICU patients. Results:A total of 263 patients were enrolled, including 54 patients with hypophosphatemia and 209 patients without. The SOFA score, LYM level and the infection rate of patients in the hypophosphatemia group were significantly higher than those in the non-hypophosphatemia group [SOFA score: 6.70±3.17 vs. 5.64±3.59, LYM (×10 9/L): 0.99±0.54 vs. 0.77±0.54, infection rate: 77.78% (42/54) vs. 59.33% (124/209), all P < 0.05], the NLR was significantly lower than that of the non-hypophosphatemia group [10.67 (7.08, 18.02) vs. 12.25 (7.25, 21.68), P < 0.05]. The length of hospital stay, ICU stay, and mechanical ventilation duration in the hypophosphatemia group were significantly longer than those in the non-hypophosphatemia group [length of hospital stay (days): 15 (11, 28) vs. 12 (6, 21), length of ICU stay (days): 10.35±7.80 vs 7.15±6.61, mechanical ventilation duration (days): 3 (0, 12) vs. 2 (0, 5), all P < 0.05]. There was no significant difference in the 28-day mortality between the hypophosphatemia group and the non-hypophosphatemia group [9.26% (5/54) vs. 11.00% (23/209), P > 0.05]. Multivariate Logistic regression analysis showed that APACHE Ⅱ score [odds ratio ( OR) = 1.188, 95% CI was 1.110-1.271], CRP ( OR = 1.016, 95% CI was 1.007-1.026), and NLR ( OR = 1.002, 95% CI was 0.996-1.008) were independent risk factors affecting the 28-day mortality of critically ill patients in ICU (all P < 0.05). ROC curve analysis showed that the AUC of serum phosphorus levels for predicting the length of hospital stay of critically ill patients in ICU > 10 days, ICU stay > 5 days, and mechanical ventilation duration > 5 days were 0.701 (95% CI was 0.632-0.770), 0.771 (95% CI was 0.691-0.852), 0.617 (95% CI was 0.541-0.692), respectively, all P < 0.01. Conclusion:Hypophosphatemia has some predictive value for the length of hospital and ICU stay and mechanical ventilation time in critically ill patients, but it cannot predict the 28-day mortality.

Meta analysis of the effects of high intensity interval training on metabolic syndrome in overweight college students / 中国学校卫生

Objective@#In this study, a meta analysis was conducted to evaluate and compare the effects of different types of interval training, such as, repeated sprint training(RST), high intensity interval training(HIIT) and sprint interval training(SIT) on body index of overweight/obese college students and the effect of moderate intensity continuous training(MICT), so as to provide a reference for taking appropriate exercise measures.@*Methods@#The data was searched and selected from the database of Web of Science, PubMed, Scopus, The Cochrane Library and CNKI, and from the articles about random research on the effects of HIIT, SIT, RST, and MICT on overweight/obese college students for the evaluation of bias risk. And Stata 16.0 software was used for Meta analysis and network analysis.@*Results@#Totally 815 samples selected from 18 articles were included in the study. Meta analysis showed that HIIT( SMD=-0.26, 95%CI =-0.52--0.00, P <0.05) and SIT( SMD=-0.39, 95%CI =-0.72--0.07, P <0.05) could make greater effects on BMI than MICT. The differences between RST and MICT were of no statistical significance( SMD=0.28, 95%CI =0.32-0.87, P >0.05). According to the SUCRA method combined with effect size, the best effect on improving MetS related physiological indexes of overweight college students was found in SIT( SUCRA =79.3), followed by HIIT( SUCRA =78.2), RST( SUCRA =56.8), and MICT( SUCRA =35.7).@*Conclusion@#Compared with MICT, high intensity interval training can greatly improve BMI of the obese/overweight college students, and could achieve the optimal effect of reducing the fat by exercising through SIT.

Effects of terlipressin combined with norepinephrine on liver function and prognosis of patients with septic shock / 国际生物医学工程杂志

Objective:To investigate the effects of terlipressin (TP) combined with norepinephrine (NE) on liver function and prognosis of patients with septic shock.Methods:From June 2018 to December 2019, 96 patients with septic shock and liver function impairment admitted to the ICU of Tianjin First Central Hospital were selected for prospective study. The patients were divided into control group( n=48) and experiment group( n=48) by randomize number table derived by computer. Based on conventional treatment, NE was used in control group, and the low dose continuous infusion of TP combined with NE was used in experiment group. Serial measurements of aspartate aminotransferase (AST), alanine aminotransferase (ALT), serum total bilirubin (TBIL), heart rate and mean arterial pressure (MAP), and blood lactic acid levels were made before the treatment and after the treatment at 24 and 48 hours. The mechanical ventilation time, intensive care unit (ICU) stay, and total length of hospital stay of the two groups were compared, and the 28-day mortality and serious adverse reactions of the two groups was also calculated. Results:The levels of ALT, AST, TBIL, heart rate and blood Lac of the two groups were significantly decreased after the treatment (all P<0.01), and the level of MAP was significantly increased (all P<0.01). Compared with the control group, the levels of 24-hour and 48-hour ALT, AST, TBIL, blood Lac of the experiment group were significantly decreased (all P<0.05), and the 48-hour level of MAP was significantly increased (all P<0.05), but there was no statistically significant difference between the two groups in the levels of 24-hour heart rate and 24-hour MAP (all P>0.05). Besides, there was no statistically significant difference between the two groups in the mechanical ventilation time, ICU stay, total length of hospital stay and the 28-day mortality (all P>0.05). And there were no serious adverse reactions such as avascular necrosis of the fingers and myocardial infraction in the two groups. Conclusions:In the treatment of septic shock, on the basic of adequate fluid resuscitation, continuous intravenous pumping of low-dose TP combined with NE can play a certain protective effect on the live, and the mechanisms of action may be mediated by stabilizing hemodynamics, reducing heart rate, reducing the level of blood Lac and improving liver perfusion, thereby protecting liver function in patients with septic shock.

Gene cloning, protein expression and examination of biological activity of chicken CD40L / 生物工程学报

To obtain chicken CD40L protein, the cDNA was prepared from chicken splenic cells and used as a template to clone and amplify CD40L by PCR. The target gene was cloned into pFastBac vector to construct a pFastBac-chCD40L donor plasmid. Recombinant plasmid was transformed into DH10Bac and recombinant Bacmid-chCD40L was obtained. The Bacmid-chCD40L plasmid was transfected into sf9 insect cells to obtain His-chCD40L protein. In addition, the target gene was cloned into pQM01 vector to construct a pQM01-chCD40L plasmid, recombinant plasmid was transfected into HEK 293T cells to obtain Strep-chCD40L protein. The chCD40L protein was purified by affinity chromatography, and the concentration of purified chCD40L protein was determined to be 0.01 mg/mL. Primary cells were isolated from the bursal tissue of 3-week old SPF chickens, and the chCD40L protein was added to the culture medium to stimulate cells. The chCD40L could bind to CD40 on B cells as examined by Western blotting, indirect immunofluorescence assay and flow cytometry, suggesting that chCD40L protein is biologically active. We successfully obtained chicken CD40L protein of biological activity, which laid the foundation in the in vitro culture of primary B lymphocytes for the isolation and diagnosis of virulent IBDV.

Predictive value of SIRT2 in the progression of sepsis to persistent inflammation-immunosuppression and catabolism syndrome / 中华急诊医学杂志

Objective:To explore the predictive value of SIRT2 in the progression of sepsis to persistent inflammation-immunosuppression and catabolism syndrome (PICS).Methods:From June 2018 to June 2019, 81 sepsis patients in Intensive Care Unit of Tianjin First Center Hospital were enrolled, and 20 healthy adult volunteers were recruited as controls. Forty-five patients who had been hospitalized for more than 14 d were selected and divided into the non-PICS group and PICS group. Blood samples were collected at 0, 24 h, 4 d, 7 d, 10 d, 14 d, 17 d, and 21 d. The levels of SIRT2, PD-1, TNF-α, IL-6, IL-10 and TGF-β were measured at different time points. In the control group, and fasting was taken only once in the morning. ROC curve was drawn and AUC was calculated to evaluate the value of SIRT2 and PD-1 in predicting sepsis progression to PICS.Results:(1) Compared with the control group, the expression of SIRT2 and PD-1 decreased at admission in the non-PICS group and PICS group ( P<0.05). Compared with the non-PICS group, the expression of SIRT2 and PD-1 in the PICS group increased at 10 and 14 d, respectively. SIRT2 in the PICS group had statistical difference at 10 d [(0.87±0.08) and (1.15±0.09), respectively; P<0.05]. PD-1 was statistical difference at 14 d between the two groups[ (0.86±0.04 )and (1.01±0.02), respectively; P<0.05]. (2) Over time, TNF-α and IL-6 in the two groups declined gradually, but IL-10 and TGF-β in the PICS group were higher than those in the non-PICS group at 10 d ( P<0.05). (3) The AUC of PD-1 was 0.766 (95% CI: 0.624-0.908), and the sensitivity and specificity were 70.8% and 81.0%, when the cut-off value was 1.01. The AUC of SIRT 2 was 0.841 (95% CI: 0.722-0.960), and the sensitivity and specificity were 79.2% and 81.0%, when the cut-off value was 1.10. Conclusions:SIRT2 expression level changes when sepsis patients enter PICS stage. SIRT2 has certain predictive value for the occurrence of PICS.

Effect of recombinant human thrombopoietin on endotoxin-induced thrombocytopenia in mice / 中华危重病急救医学

Objective To investigate the effect of recombinant human thrombopoietin (rhTPO) on thrombocytopenia (TCP) induced by endotoxin lipopolysaccharide (LPS) in mice. Methods Sixty male C57BL/6 mice were divided into normal saline (NS) control group (NS group), sepsis-induced TCP model group (LPS group) and rhTPO treatment group (LPS+rhTPO group) by random number table with 20 mice in each group. Sepsis-induced TCP model was reproduced by one intraperitoneal injection of LPS 30 mg/kg, and the mice in NS group were given the same amount of NS. In LPS+rhTPO group, 2.7 kU/kg rhTPO was subcutaneously injected into mice immediately after intraperitoneal injection of LPS, once every 24 hours. The mice in NS group and LPS group were injected subcutaneously with the same amount of NS. The observation period of each group lasted for 72 hours. The inner canthus blood was harvested before and every 24 hours after modeling, and the platelet count (PLT) was measured by animal blood cell counter. The eyeball blood of mice was harvested at 72 hours after modeling, and the proportion of CD61+CD62p+ cells in platelet-rich plasma was detected by flow cytometry, by which the platelet activation was reflected. Lung and spleen tissues of mice were harvested, and the positive expression of CD41 was determined by immunohistochemistry, by which the platelet sequestration in organs was reflected. Bone marrow cells from unilateral femur of mice were harvested, and the proportion of CD41+CD61+ cells was determined by flow cytometry to reflect the proliferation of bone marrow megakaryocytes. Results There was no significant difference in PLT among the groups before modeling. With the extension of the time after modeling, PLT in LPS group was decreased continuously, and increased slightly at 72 hours, but it was still significantly lower than that in NS group (×109/L: 308.60±21.70 vs. 1 152.72±50.27, P < 0.05); PLT in LPS+rhTPO group was increased continuously with the extension of modeling time, and it was significantly higher at 72 hours than that in LPS group (×109/L: 926.78±48.85 vs. 308.60±21.70, P < 0.05). At 72 hours after modeling, the proportion of CD61+CD62p+ cells in platelet-rich plasma of LPS group was significantly higher than that of NS group [(25.07±2.55)% vs. (4.17±0.38)%, P < 0.05], while the value in LPS+rhTPO group was significantly lower than that of LPS group [(15.92±1.26)% vs. (25.07±2.55)%, P < 0.05]. The proportion of CD41+CD61+ cells in bone marrow megakaryocytes of LPS group was significantly higher than that of NS group [(11.84±0.80)% vs. (3.60±0.42)%, P < 0.05], and the proportion of CD41+CD61+ cells in LPS+rhTPO group was significantly higher than that in LPS group [(30.96±2.49)% vs. (11.84±0.80)%, P < 0.05]. Immunohistochemistry showed that the positive expressions of CD41 in lung and spleen tissues of LPS group increased significantly than NS group [A value: 828.94±119.30 vs. 447.09±16.19 in lung tissue, (280.15±16.71)×103 vs. (0.65±0.26)×103 in spleen tissue, both P < 0.05], while the positive expressions of CD41 in lung and spleen tissues of LPS+rhTPO group decreased significantly than LPS group [A value: 542.78±2.95 vs. 828.94±119.30 in lung tissue, (129.40±13.49)×103 vs. (280.15±16.71)×103 in spleen tissue, both P < 0.05]. Conclusion The rhTPO in endotoxin-induced TCP may stimulate the proliferation of bone marrow megakaryocytes, inhibit platelet activation and affect platelet sequestration in organs, so as to increase platelet levels.

Effect of recombinant human thrombopoietin on endotoxin-induced thrombocytopenia in mice / 中华危重病急救医学

Objective@#To investigate the effect of recombinant human thrombopoietin (rhTPO) on thrombocytopenia (TCP) induced by endotoxin lipopolysaccharide (LPS) in mice.@*Methods@#Sixty male C57BL/6 mice were divided into normal saline (NS) control group (NS group), sepsis-induced TCP model group (LPS group) and rhTPO treatment group (LPS+rhTPO group) by random number table with 20 mice in each group. Sepsis-induced TCP model was reproduced by one intraperitoneal injection of LPS 30 mg/kg, and the mice in NS group were given the same amount of NS. In LPS+rhTPO group, 2.7 kU/kg rhTPO was subcutaneously injected into mice immediately after intraperitoneal injection of LPS, once every 24 hours. The mice in NS group and LPS group were injected subcutaneously with the same amount of NS. The observation period of each group lasted for 72 hours. The inner canthus blood was harvested before and every 24 hours after modeling, and the platelet count (PLT) was measured by animal blood cell counter. The eyeball blood of mice was harvested at 72 hours after modeling, and the proportion of CD61+CD62p+ cells in platelet-rich plasma was detected by flow cytometry, by which the platelet activation was reflected. Lung and spleen tissues of mice were harvested, and the positive expression of CD41 was determined by immunohistochemistry, by which the platelet sequestration in organs was reflected. Bone marrow cells from unilateral femur of mice were harvested, and the proportion of CD41+CD61+ cells was determined by flow cytometry to reflect the proliferation of bone marrow megakaryocytes.@*Results@#There was no significant difference in PLT among the groups before modeling. With the extension of the time after modeling, PLT in LPS group was decreased continuously, and increased slightly at 72 hours, but it was still significantly lower than that in NS group (×109/L: 308.60±21.70 vs. 1 152.72±50.27, P < 0.05); PLT in LPS+rhTPO group was increased continuously with the extension of modeling time, and it was significantly higher at 72 hours than that in LPS group (×109/L: 926.78±48.85 vs. 308.60±21.70, P < 0.05). At 72 hours after modeling, the proportion of CD61+CD62p+ cells in platelet-rich plasma of LPS group was significantly higher than that of NS group [(25.07±2.55)% vs. (4.17±0.38)%, P < 0.05], while the value in LPS+rhTPO group was significantly lower than that of LPS group [(15.92±1.26)% vs. (25.07±2.55)%, P < 0.05]. The proportion of CD41+CD61+ cells in bone marrow megakaryocytes of LPS group was significantly higher than that of NS group [(11.84±0.80)% vs. (3.60±0.42)%, P < 0.05], and the proportion of CD41+CD61+ cells in LPS+rhTPO group was significantly higher than that in LPS group [(30.96±2.49)% vs. (11.84±0.80)%, P < 0.05]. Immunohistochemistry showed that the positive expressions of CD41 in lung and spleen tissues of LPS group increased significantly than NS group [A value: 828.94±119.30 vs. 447.09±16.19 in lung tissue, (280.15±16.71)×103 vs. (0.65±0.26)×103 in spleen tissue, both P < 0.05], while the positive expressions of CD41 in lung and spleen tissues of LPS+rhTPO group decreased significantly than LPS group [A value: 542.78±2.95 vs. 828.94±119.30 in lung tissue, (129.40±13.49)×103 vs. (280.15±16.71)×103 in spleen tissue, both P < 0.05].@*Conclusion@#The rhTPO in endotoxin-induced TCP may stimulate the proliferation of bone marrow megakaryocytes, inhibit platelet activation and affect platelet sequestration in organs, so as to increase platelet levels.

Effect of alveolar macrophages phagocytosis on prognosis in patients with acute respiratory distress syndrome caused by abdominal infection: a multicenter study / 中华危重病急救医学

Objective To investigate the effects of alveolar macrophage phagocytosis on prognosis in patients with acute respiratory distress syndrome (ARDS) caused by abdominal infection. Methods ARDS patients caused by severe intra-abdominal infection admitted to intensive care unit (ICU) of Tianjin Fourth Central Hospital, Tianjin Nankai Hospital, Tianjin First Central Hospital and Tianjin Fifth Central Hospital from June 2016 to March 2018 were enrolled. The gender, age, acute physiology and chronic health evaluationⅡ(APACHEⅡ) within 24 hours of admission, neutral red phagocytosis and alkaline phosphatase activity of macrophages in bronchoalveolar lavage fluid, the length of ICU stay, total hospitalization time, hospitalization expenses, and prognosis were recorded. According to the prognosis, the patients were divided into death group and survival group, and the parameters were compared between the two groups. Pearson test was used to analyze the correlation between neutral red phagocytosis function of macrophages and alkaline phosphatase activity and other indicators. The prognosis was analyzed by binary Logistic regression combined with neutral red phagocytosis and alkaline phosphatase activity in patients, and the predictive value of both subjects on prognosis was analyzed by the receiver operating characteristic (ROC) curve. Results Twenty patients were enrolled in the study, with 8 in the death group and 12 in the survival group. Compared with the survival group, the death group was older (years old: 58.50±14.86 vs. 46.67±13.40), APACHEⅡ score was higher (21.50±3.93 vs. 13.58±4.12), neutral red phagocytosis ability and alkaline phosphatase activity of alveolar macrophages were significantly decreased (A value:0.265±0.050 vs. 0.338±0.016; μmol/L: 12.06±1.24 vs. 17.96±3.90), and the length of ICU stay was significantly longer (days: 22.00±14.59 vs. 11.50±3.17), hospitalization cost was significantly increased (10 thousand Yuan:24.17±11.02 vs. 13.44±3.53), the total hospitalization time was shorter (days: 25.25±15.01 vs. 35.67±8.58), and the difference was statistically significant (all P < 0.05). There was no significant difference in gender between the survival group and the death group [male (case): 8 vs. 6, P > 0.05]. The neutral red phagocytosis ability of alveolar macrophages in ARDS patients caused by abdominal infection was negatively correlated with age, APACHEⅡ score and the length of ICU stay (r value was -0.328, -0.572, -0.809, respectively, all P < 0.05); alkaline phosphatase activity was negatively correlated with age, APACHEⅡ score, the length of ICU stay and hospitalization expenses (r value was -0.334, -0.583,-0.470, -0.517, respectively, all P < 0.05). Binary Logistic regression analysis showed that neutral red phagocytosis [odds ratio (OR) = 0.596, 95% confidence interval (95%CI) = 0.212-0.997] and alkaline phosphatase activity (OR = 0.573, 95%CI = 0.339-0.968) were the influencing factors of prognosis (both P < 0.05). ROC curve analysis showed that the AUC of neutral red phagocytosis ability for prognosis of ARDS patients caused by abdominal infection was 0.948, and the sensitivity and specificity were 91.7% and 87.5% when the off-cut value was 0.317. The AUC of alkaline phosphatase for the prognosis of ARDS patients caused by abdominal infection was 0.813; when the cut-off value was 19.72 μmol/L, the sensitivity was 75.0%, and the specificity was 87.5%. Conclusion The alveolar macrophage phagocytosis dysfunction in ARDS patients caused by severe abdominal infection was not only related to the severity of the disease, but also increased the medical burden of patients, and significantly affected the mortality of such patients.

Clinical characteristics and prognosis of acute kidney injury in elderly patients with sepsis / 中华危重病急救医学

Objective To analyze the incidence of acute kidney injury (AKI) in elderly patients with sepsis, compare the clinical characteristics and prognosis between AKI and non-AKI elderly patients with sepsis, and to investigate the impact of classification of AKI and renal replacement therapy (RRT) on the outcome of elderly patients with sepsis. Methods The clinical data of 490 septic patients over 65 years old, admitted to intensive care unit (ICU) of Tianjin First Center Hospital from April 1st, 2016 to December 31st, 2018 were retrospectively analyzed. The patients were divided into two groups according to those with or without AKI. The clinical characteristics of patients were compared, and subgroup analysis of elderly septic patients with AKI was performed according to Kidney Disease:Improving Global Outcomes (KDIGO) staging criteria and whether RRT was performed, to observe the effects of AKI staging and RRT on the prognosis of elderly septic patients with AKI. Multivariate Cox regression analysis was used to screen the risk factors of death in elderly patients with sepsis associated AKI. Results ① A total of 490 septic elderly patients were enrolled, including 249 patients with AKI and 241 patients without AKI, with the AKI incidence of 50.8%. Compared with non-AKI group, the patients in AKI group were older (years old: 72.0±7.2 vs. 68.8±5.1), acute physiology and chronic health evaluation Ⅱ (APACHEⅡ) score and sequential organ failure assessment (SOFA) score were evidently higher (23.1±6.1 vs. 22.0±3.7, 9.4±3.8 vs. 6.1±3.5); the duration of mechanical ventilation [days: 7.0 (5.0, 10.0) vs. 6.0 (3.0, 9.0)], length of ICU stay [days: 12.0 (7.0, 15.0) vs. 7.0 (4.0, 13.0)] and total length of hospital stay [days: 15.0 (10.0, 21.5) vs. 12.0 (7.0, 15.0)] were longer, and ICU mortality and 28-day mortality were evidently higher [22.9% (57/249) vs. 14.1% (34/241), 36.1% (90/249) vs. 24.5% (59/241), all P < 0.05]. ② According to KDIGO staging, 93 patients were in stage 1, 70 in stage 2 and 86 in stage 3 of AKI. The rate of RRT was increased with increase in KDIGO staging [14.0% (13/93), 30.0% (21/70), 88.4% (76/86)], the duration without mechanical ventilation within 28 days was shortened [days: 20.0 (0, 23.0), 8.0 (0, 20.5), 8.0 (0, 13.0)], the rate of kidney recovery was decreased [71.0% (66/93), 51.4% (36/70), 37.2% (32/86)], meanwhile, the ICU and 28-day mortality was increased [12.9% (12/93), 38.6% (27/70), 20.9% (18/86), and 26.9% (25/93), 35.7% (25/70), 46.5% (40/86), all P < 0.05]. ③ 110 elderly septic patients with AKI were treated with RRT, and 139 without RRT. Compared with non-RRT group, the ratio of mechanical ventilation in RRT group was lowered [46.4% (51/110) vs. 68.3% (95/139)], the duration without mechanical ventilation within 28 days [days: 18.0 (0, 23.0) vs. 10.0 (0, 13.0)], the length of ICU stay [days: 13.0 (12.0, 17.9) vs. 10.0 (6.0, 14.0)] and the total length of hospital stay [days: 22.5 (15.0, 46.0) vs. 16.0 (12.0, 23.0)] were prolonged, and the 28-day mortality was evidently increased [50.0% (55/110) vs. 25.2% (35/139), all P < 0.01], however, no significant difference in ICU mortality was found [27.3% (30/110) vs. 19.4% (27/139), P > 0.05]. ④ Cox regression analysis showed that SOFA score [relative risk (RR) = 1.214, 95% confidence interval (95%CI) = 1.117-1.319], KDIGO stage (RR = 4.077, 95%CI =1.850-8.982), vasoactive substance usage (RR = 2.896, 95%CI = 1.502-5.584), and mechanical ventilation (RR = 5.787, 95%CI = 1.512-22.156) were the risk factors of 28-day mortality in elderly septic patients with AKI (all P < 0.05). Conclusions The incidence of AKI for elderly septic patients with AKI was about 50%, who had a worse prognosis as compared with non-septic AKI patients. The higher the stage of KDIGO, the higher the mortality in elderly septic patients with AKI was. RRT can decrease the rate of mechanical ventilation, whereas, it may not improve the prognosis of elderly septic patients with AKI.

Expression of IL-18 in sepsis-associated thrombocytopenia and its clinical significance / 实用医学杂志

Objective To investigate the expression of IL-18 in severe sepsis and sepsis-associated throm-bocytopenia and explore its clinical significance.Methods Real-time polymerase chain reaction was applied to de-tect the expression of IL-18 miRNA in 28 samples of sepsis-associated thrombocytopenia patients and 32 samples of severe sepsis. The enzyme-linked immunosorbent assay was applied to detect the concentration of IL-18 in their plasma. Results The miRNA expression of IL-18 in the sepsis-associated thrombocytopenia group was higher than in the severe sepsis group(P=0.015).The concentration of IL-18 in the severe sepsis-associated thrombocyto-penia group were also higher than those in the severe sepsis group(P=0.034). Conclusion The expression level of IL-18 is related with the severity of thrombocytopenia in patients with sepsis and is likely to play an important role in the pathogenesis of sepsis-associated thrombocytopenia.

Fabrication and pre-clinical application of patient-specific 3D silicone rubber bolus for chest wall / 中华放射肿瘤学杂志

Objective To apply 3D printing technology to fabricate patient-specific silicone tissue compensators for the chest wall and compare the advantages and clinical characteristics between conventional bolus and 3D-printed PLA materials. Methods The chest wall data of two breast cancer patients undergoing mastectomy were obtained based upon the CT images. A patient-specific 3D printing silicone rubber bolus (3D-SRB) was designed and fabricated. The conformability of 3D-SRB,3D-PLA and conventional bolus to the chest wall were validated. Ecipse8. 6 planning system was adopted to statistically compare the dosimetric parameters of virtual plan with those after using three tissue compensators. Results The 3D-SRB was successfully designed and fabricated with a similar hardness to conventional bolus. During the process of validating conformability and radiotherapy planning,3D-SRB and 3D-PLA were superior to conventional bolus in terms of conformability to chest wall and planning dosimetric distribution.3D-SRB was advantageous in repeatability, conformability and comfortable experience compared with 3D-PLA. Regarding dosimetric parameters,3D-SRB yielded the highest repeatability with the virtual plan, followed by 3D-PLA and conventional bolus. Conclusion It is applicable to utilize 3D-SRB as the patient-specific compensators for the chest wall,which is of significance in clinical practice.

Association of the immature platelet fraction with sepsis diagnosis and severity / 中华急诊医学杂志

Objective To explore the relationship between immature platelet fraction(IPF) with severity of sepsis and prognosis in patients with septic shock.Methods A total of 40 patients admitted to intensive care units of Tianjin First Central Hospital from June 2016 to June 2017 were enrolled.Of them,10 patients contracted non-sepsis infected,13 patietns with septic shock,and 17 patients with non-complicated sepsis.Ten healthy subjects were recruited as control groups from Tianjin Medical University.IPF and immature reticulocyte fraction (IRF) were detected,and SOFA and APACHE Ⅱ scores were calculated,and clinical findings of all groups were recorded.The differences in IPF and IRF between the groups were analyzed.The relationship between the IPF and SOFA score was studied,and the role of IPF in the diagnosis of septic shock was evaluated.Statistical methods include t test,MarmWhitney test,Spearman correlation analysis,and ROC procedure,and P＜0.05 was considered significant.Results Significantly higher IPF level was observed in patients with sepsis than that in patients with nonsepsis infection.(6.25 + 2.92) vs.(2.49 ± 1.03),P＜0.01.Significantly higher IPF level was observed in patients with septic shock than that in patients with non-complicated sepsis(4.71 ± 1.79) vs.(8.25 ± 2.94),P＜0.01.IPF correlated with sepsis severity scores (7.41 ± 3.51) vs.(4.5 ± 1.7),P=0.005;r=0.58,P=0.001.This study presented the highest diagnostic accuracy for the presence of sepsis by all studied clinical and laboratory parameters (AUC=0.78,P=0.01).Conclusion IPF levels could be used as a biomarker for diagnosis and severity of sepsis.

Effects of common hemoglobin variants on 4 ion exchange high performance liquid chromatography methods based HbA1c measurements / 中华检验医学杂志

Objective To evaluate effects of 7 common hemoglobin variants on HbA 1c measurements using 4 ion exchange high performance liquid chromatography methods .Methods Ninety five samples with hemoglobin variants were collected from January 2017 to February 2018 during HbA1c measurements in laboratary medicine of peking university shenzhen hospital .Samples with 7 common hemoglobin variants were measured using Sebia Capillary 2 Flex Piercing, Bio-Rad D-10, Arkray HA8180V, Tosoh G8, and MQ6000 Plus, respectively.Effects of 7 common hemoglobin variants on HbA 1c measurements by the 4 methods were analyzed using Capillary 2 Flex Piercing as a comparative method .All statistical analyses were carried out using SPSS software version 19.0 .Mean bias were calculated for samples with hemoglobin variants , box plot was established to display bias distribution .Results Hb New York showed no interference on the 4 HPLC mechods although Hb New York could not be detected .D-10 could detect 6 Hb variants, and showed clinically significant interference for Hb J-Bangkok, Hb G-Coushatta, and Hb G-Taipei.HA-8180V fast mode yielded no HbA1c values for Hb J-Bangkok, Hb G-Coushatta, and Hb G-Taipei.Hb E, Hb Q-Thailand, and Hb G-Honolulu produced significant negative biases for HA-8180V.G8 standard mode could detect 1 Hb variant, and showed significant negative biases for six Hb variants .MQ6000 Plus could separate six Hb variants , only Hb G-Coushatta and Hb G-Taipei produced significant negative biases for the system . Conclusions Some common hemoglobin variants can interfere with HbA 1c determination by the most popular methods in South China , which may lead to erroneous HbA 1c values.

Generation and characterization of monoclonal antibodies against chicken interleukin 4 / 生物工程学报

To develop monoclonal antibodies (McAbs) against chicken interleukin 4 (chIL-4), we subcloned the mature chIL-4 gene into prokaryotic expression vectors pET-28a and pGEX-6P-1, then expressed and purified the recombinant proteins. We immunized BALB/c mice with the purified His-chIL-4 protein and fused the murine splenocytes with SP2/0 after 4 times of immunization. We used the GST-chIL-4 protein as a coating antigen to establish an indirect ELISA to screen positive clones. After screening and 3 rounds of cloning process, we obtained 3 hybridomas that stably secreted McAbs against chIL-4, and named 1G11-3B, 2E5-3D, and 1G11-5H. The isotypes of these McAbs were all IgG1 and the dissociation constant (Kd) of these McAbs were 1.79×10⁻⁹, 1.61×10⁻⁹, and 2.36×10⁻⁹, respectively. These McAbs specifically bound to chIL-4 expressed by either prokaryotic or eukaryotic system as determined by Western blotting and indirect immunofluorescence assay. The binding domains of chIL-4 recognized by 1G11-3B, 2E5-3D, and 1G11-5H were located between aa 1-40, 80-112, and 40-80, respectively, as determined by Western blotting. These McAbs would help to detect chIL-4 and to elucidate the biological roles of chIL-4 in immune responses.

Timing of tracheostomy in ICU patients with mechanical ventilation support / 中华急诊医学杂志

Objective To assess the influence of timing of tracheostomy performed on ICU patientswith mechanical ventilation support for long-term.Methods A retrospective study was carried out in 94 patients under mechanical ventilation support with tracheostomy from January 2012 to October 2014.The patients were divided into early stage group (group A) in which the tracheostomy was done within 7 days after endotracheal intubation and late stage group (group B) in which the tracheostomy was performed at above 7 days after endotracheal intubation.The differences in lengths of mechanical ventilation support (MVS),ICU stay,and hospital stay,incidence of ventilator-associated pneumonia (VAP) and mortality were compared between two groups using nonparametric statistics.Results Compared with group B,there were statistically significant reduction in duration of mechanical ventilation (7d vs.17 d;P ＜ 0.05),shorter length of ICU stay (10 d vs.19 d;P ＜ 0.05),and lower incidence of VAP (21.05％ vs.36.84％;P ＜ 0.05) in group A.There were no significant differences in hospital stay and mortality between two groups (P ＞0.05).There was a correlation between the duration of mechanical ventilation and timing of tracheostomy (R2 =0.680) and a correlation between the length of ICU stay and the timing of tracheostomy (R2 =0.662) was found.Conclusions Early tracheostomy has a significant positive impact on critically ill patients hospitalized in this ICU.These results support the tendency to balance the risk-benefit analysis in favor of early tracheostomy.

Experience of Laparoscopy and Colonoscopy Cooperative Surgery in Treatment of Colon Tumors / 中国微创外科杂志

Objective To investigate the result of colonoscopic positioning in laparoscopic treatment of colon tumors . Methods A retrospective analysis was made in 16 cases of colon tumors treated with laparoscopic colon resection from January 2014 to November 2015.During the operation, the lesion could not be locatized and the resection range could not be determinated because the serous layer was not involved or the lesion was located in the lateral abdominal wall .Transanal colonoscopic positioning of the lesion with light transmission method was carried out . Results The pathological changes were found under colonoscopy in all the 16 cases and the laparoscopic resection was successfully completed without conversion to open operation .No missed resection or incorrect resection occurred.The operation time was (112.5 ±31.0) min and the blood loss was (22.8 ±11.2) ml.Among 9 cases of preoperative pathological diagnosis of benign tumors , there were 6 cases of malignant tumors after surgery , including 4 cases of moderately differentiated adenocarcinoma , 1 case of moderate to severe atypical hyperplasia with cancerous lesions , and 1 case of moderately differentiated adenocarcinoma with mucous adenocarcinoma .A total of 15 patients were discharged from hospital at 12-24 days after the operation , whereas 1 patient was dismissed 1 month after the operation due to anastomotic fistula .The 16 patients were followed up for 3 months with no recurrence under colonoscopic examiantion . Conclusions During laparoscopic colon resection , if the tumor can not be locolized , colonoscopic positioning can help prevent missed resection or incorrect resection , which increases the indication and safety of laparoscopic and colonoscopic minimally invasive treatment .It has good clinical application value and deserves further promotion .

A multicenter study of targeted surveillance at Intensive Care Unit acquired MRSA infection in Tianjin district / 中华急诊医学杂志

Objective To study the incidence and risk factors of infection caused by methicillinresistant staphylococcus aureus (MRSA) with a targeted surveillance at intensive care unit (ICU) acquired MRSA infection in Tianjin area.Methods A prospective multi-center observational analysis of consecutive patients admitted to 15 adult ICUs from March 1,2012 through March 31,2014 was carried out.The ICUs were divided into four groups according to the type of the ICU.All of the patients were cared for with routine MRSA surveillance.A number of risk markers and prognostic factors were recorded.The risk factors contributing to ICU acquired MRSA were evaluated using a logistic regression model.Comparison of survival between groups was analyzed with Kaplan-Meier method.Results A total of 1 787 patients were enrolled,and 144 cases of them were MRSA infections.The patients with MRSA infection were significantly older than those with non-MRSA infection (P =0.043),length of ICU stay,length of antimicrobial therapy,the history of repeated administration of antibiotics in recent days,history of operation in the past five years,history of MRSA infection or colonization,frequent application of and the overall length of time for mechanical ventilation and central venous catheter and catheter-associated infection were significantly higher than those with non-MRSA infection.The survival rate of patients with non-MRSA infection were higher than those with MRSA infection (x2 =9.23,P =0.004).The rate of MRSA infection and MRSA colonization in 2013 were significantly lower than that in 2012,because the rate of hand hygiene rule execution and bacterial clearance rate were significantly higher in 2013.Multivariate Logistic regression analysis demonstrated that advanced age (OR =1.05,95％ CI:1.009-1.086),length of ICU stay (OR =1.05,95％ CI:1.01-1.08),history of MRSA infection or colonization (OR =1.33,95％ CI:1.82 -3.27),glucocorticoid therapy (OR =2.85,95％ CI:1.18-6.91),antacid medicine (OR =4.92,95％ CI:1.18-20.58),history of recent or repeated application of antibiotics (OR =3.26,95％ CI:1.06-4.59) catheter-associated infections (OR =2.22,95％ CI:1.08-4.59) were associated with ICU acquired MRSA infections.Conclusions Performing the rule of hand hygiene strictly as well as strengthening prevention and control of MRSA infections can effectively reduce the incidence of ICU acquired MRSA infections.The advanced age,length of ICU stay,history of MRSA infection or colonization,glucocorticoid therapy,antacids medicine,history of recent or repeated application of antibiotics,catheterassociated infections were independent risk factors of ICU acquired MRSA infections.