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Objective To investigate the constitution and drug resistance status of the major pathogens in mechanically ventilated patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD),so as to provide evidences for clinical treatment.Methods From patients with AECOPD undergoing mechanical ventilation in EICU or ICU of the Second Affiliated Hospital of Wenzhou Medical University from January 2016 to December 2017,various specimens were collected for identification and drug susceptibility testing of pathogens;the clinical data and test results were analyzed.Results A total of 104 patients were included in the study.Total of 163 strains of pathogens were isolated from the specimens,of which gram-negative pathogens were the most common,accounting for 74.8% of any pathogens.Drug resistance analysis showed that gram-negative pathogens were severely resistant to third generation cephalosporins,and were more sensitive to Sulperazon and carbapenems;gram-positive cocci were more sensitive to vancomycin and linezolid.Fungi were more sensitive to amphotericin B and 5-fluorocytosine.Most of the pathogens were multi-resistant.Conclusions In selecting antibiotics for the treatment of critical patients with AECOPD,emphasis should be placed on bacterial culture and drug susceptibility testing,so as to reduce irrational drug use and the emergence of drug-resistant strains or superinfection.
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Objective To explore the role of receptor for advanced glycation end products (RAGE) in HMGBl-mediated CD4+ T cells differentiation to Th1/Th2.Methods CD4+ T lymphocytes isolated from the spleens of male BALB/C mice by magnetic beads were suspended in RPMI-1640 with 10% FCS in 2× 106cell/well on 96-well cell culture plates in vitro.The cells were randomly divided 4 groups according to concentration of HMGB1 treatment:control group,10 ng/mL group,100 ng/mL group,1 000 ng/mL group after stimulation with ConA in 3 μg/mL for 12 hours.IL-4 and IFN-γ levels in culture supernatants were quantitated by ELISA kits after HMGB1 stimulation for 12,24,and 48 h.According to the results,cultured cells were exposed to HMGB1 in 100 ng/mL for 24 h in the following experiments.The cells were randomly divided into 4 groups:control group,A group,B group,C group,and each group were cultured with ConA in 3 μg/mL for 24 h.The cells of control group and other three groups were stimulated with PBS or 100 μg/L HMGB1 for 24 h.The cells of B,C groups were incubated with 1/200 diluted 5 μg/L anti-RAGE Abs (anti-bodies) or PBS for 2 h before HMGB1 stimulation.The cell suspension was obtained to detect the levels of IL-4 and IFN-γ by EILSA and the protein levels and mRNA expressions of RAGE,CATA-3 were detected by western-blot and real-time fluorescent quantitative PCR,respectively.ResuRs Compared with control group,CD4+ T cells incubated with increasing concentrations of HMGB1 (100,1 000 ng/mL) for 24 h resulted in a decrease in IFN-γ/IL-4 ratio (P<0.05).When CD4+ T cells were exposed to 100 ng/mL HMGB1 for 12 h,IFN-γ/IL-4 ratio was markedly increased.However,CD4+ T cells treated with 100 ng/mL HMGB1 for 24,48 h,IFN-γ/IL-4 ratio was markedly inhibited (P<0.05).Compared with control group,protein levels and mRNA expressions of RAGE and GATA3 of cells in A group were significantly increased (P<0.05),and IFN-γ/IL-4 ratio of cell suspension in A group and B group was significantly down-regulated (P<0.05).Compared with A group,IFN-γ/IL-4 ratio of cell suspension in C group was increased (P<0.05),and expression of GATA3 mRNA was down-regulated (P<0.05).Compared with A group,protein level of RAGE of cells in C group was significantly down-regulated (P<0.05),but protein level of RAGE of cells in C group was still increased compared with control group (P<0.05).Conclusion Th1/Th2 differentiation induced by HMGB1 on CD4+ T lymphocytes in vitro was at least partly mediated by over-activating RAGE/GATA3 pathway.
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Objective To investigate the effect of continuous renal replacement therapy ( CRRT) on the reg-ulation of Treg/Th17 in patients with severe sepsis,and related inflammatory factors IL-6,IL-17,IL-10,and TNF-α.Methods 60 patients with severe sepsis were randomly divided into two groups,30 cases in each group.The control group received conventional treatment,and the observation group was treated with CRRT on the basis of the control group.Flow cytometry was used to detect the levels of Tregs and Th17 cells,and IL-6,IL-10,IL-17 and TNF-αwere detected by ELISA method.At the same time,the APACHEII score,ICU length of hospital stay were observed and recorded.Results After treatment,APACHE Ⅱscore,ICU length of hospital of the observation group were lower than the control group,there were statistically significant differences(t=4.258,t=4.518,all P0.05].Conclusion CRRT can not only remove the inflammatory mediators of abnormal expression,improve the function of T cells,but also can maintain the balance between Th17 and Treg,improve the immune disorders,and improve the prognosis of sepsis.
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Objective To investigate the serum high mobility group box-1 protein level of children's patients with a-cute traumatic brain injury and it's expression changes and clinical significance. Methods A total of 87 children's patients with acute traumatic brain injury were randomly divided into three groups: severe group (≤8 scores), moderate group(9 to 12 scores) and minor group(13 to 15 scores) according to the patients' Glasgow Coma Scale on admission. Their venous blood were drawn at 3 h, 6 h, 12 h, 24 h, 48 h after injury and the serum HMGB1 were determined by en zyme linked immunosorbent assay(ELISA). The serum HMGB1 concentration were compared between different groups, survivors and non-survivors in 28 days, patients who developed acute lung injury(ALI) or sepsis and who did not deve-lope ALI or sepsis in severe injury group. Results Compared with the minor and moderate group, the serum HMGB1 level in severe group were significantly elevated at each time points(P<0.01). At 6 h, 12 h, 24 h, 48 h time points, the serum HMGB1 level in the moderate group increased significantly compared with the minor group (P<0.01). The serum HMGB1 levels at 24 h after injury were higher in non-survivors compared with survivors (P<0.01). The serum HMGB1 levels in those patients who developed acute lung injury (ALI) or sepsis were higher than who did not develope ALI or sepsis (P<0.01). Conclusion The serum HMGB1 concentration is increased early and may be a ‘alarm signal'prog-nosticating prognosis and complication after acute traumatic brain injury in children.
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Objective To evaluate of impact of thalidomide on CD4 +CD25 +T lymphocytes in patients with acute myeloid leukemia and its clinical curative effect.Methods 71 cases of patients with AML were randomly divided into the control group and the experiment group, and 31 healthy people were selected to be the normal group.The experiment group and the control group patients were treated with the same chemotherapy, the experiment group was treated with thalidomide.The levels of CD4 +CD25 +T lymphocyte, CD3 +T lymphocyte, CD4 +T lymphocyte, CD8 +T lymphocyte, ratio of CD4 +/CD8 +and NK cells were detected at before treatment, 10 to 14d after treatment, complete remission 6 months follow-up.In normal group, the same index was detected before and after chemotherapy, and 10-14 d.The clinical curative effect of the experiment group and the control group were observed.Results The effective rate of the experiment group was higher than of the control group(P<0.05); The levels of CD4 +CD25 +T lymphocyte in the experiment group and control group were significantly higher than in the normal group(P<0.05), the two groups decreased compared with the control group, the level of CD4 +CD25 +T lymphocyte in the experiment group was significantly decreased(P<0.05).And with thalidomide treatment, the experiment group in the CD3 +T lymphocytes, CD4 +T lymphocytes and ratio of CD4 +/CD8 +, NK cells were significantly higher than the control group ( P<0.05 ) .Conclusion Thalidomide can improve both the immunity cell function and the clinical efficiency in patients with AML.The mechanism is related to reduce the level of CD4 +CD25 +T lymphocyte.
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Objective To explore the viral etiology spectrum of different age and different seasons for neo-natal pneumonia.Methods Medical records of 1 073 cases of neonatal pneumonia in our hospital were retrospectively analyzed,line direct immunofluorescence assay were used to detect nasopharyngeal secretions of newborns,the test results were statistically analyzed.Results In 1 073 cases with neonatal pneumonia,406 cases were detected positive with virus infected,the positive rate was 37.8%,7 cases were mix infected.334 cases were infected by respiratory syncytial virus(RSV),which had the highest detection rate,accounting for 82.3%;RSV infection rate in 1 -6 month baby was 36.4%,which was higher than the >6 -12 month -old baby with RSV infection rate 26.5%,the difference was statistically significant(χ2 =12.25,P <0.05);RSV infection rate in winter and spring group was 39.7%,which was significantly higher than that in autumn and winter group(13.6%);PIV3 infection rate in winter and spring group was 1.8%,significantly lower than that in autumn and summer group(9.1%),the difference was statistically signifi-cant(χ2 =31.27,P <0.05 ).Conclusion RSV is the most common viral in neonatal pneumonia,more attention should be payed to RSV infection control in small babies and at winter and spring,pay attention to PIV3 infection at autumn and summer.
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Objective To observe the dynamic changes of high mobility group protein B1 ( HMGB1 )expression in the lung of rats with Vibrio vulnificus sepsis so as to unravel the role of HMGB1 in lung injury.Methods Sixty rats of clean grade were randomly divided into normal control group ( A group, n = 10) and Vibrio vulnificus sepsis group (B group, n =50). Sepsis model was made in rats with subcutaneous injection of Vibrio vulnificus with concentration of 6 × 108 cfu/ml in dose of 0. 1 ml/100 g into left lower limb.The rats of group B were sacrificed 1 h, 6 h, 12 h, 24 h and 48 h after infection for taking lung tissues to detect the water content of lung and to observe the histopathological changes in lung under light microscope.The expression of HMGB1 mRNA and the level of HMGB1 protein in the lungs were detected by RT-PCR and Western blot, respectively. Data were analysed with ANOVA and LSD method for comparison between groups, and P <0.05 was considered statistically significant. Results Compared with the group A (0.652±0. 177), the expressions of HMGB1 mRNA in lung of rats of group B were significantly higher in 12 hours (1. 161 ±0.358, P=0.013), 24 hours (1.679 ±0.235, P =0.000) and 48 hours (1.258 ±0.274, P=0.004) and reached the peak in 24 h. Compared with group A (0.594 ±0. 190), the level of HMGB1 protein in rats of group B 6 h after infection ( 1. 408 ± 0. 567, P = 0. 026) was significantly increased (P<0.05), and it reached peak in 24 h (2.415 ± 1.064, P =0.000) after infection. Compared with group A (0.699 ± 0.054), the lung water contents in rats of group B were significantly increased in 6 h (0.759±0.030, P=0.001), in 12 h (0.767 ±0.023, P =0.000), in 24 h (0.771 ±0.043, P=0.000) and in 48 h (0.789 ±0.137, P=0.000) after infection. Compared with group A, the pathological changes in the lung of rats in group B showed clearly marked pulmonary vascular congestion, interstitial edema and inflammatory cell infiltration, and those changes became more and more serious until alveolar sacs entirely collapsed and the boundaries of the alveolar septa could not be clearly identified in 48 h. Conclusions Vibrio vulnificus sepsis leads to the lung injury of infected rats, and the increase in the expression of HMGB1 mRNA in lung might be one of the mechanisms of lung injury in rats with Vibrio vulnificus sepsis.