Objective To construct the mouse NF-κB P65 subunit expression plasmid, and identify its biological activity. Methods NF-κB P65 siRNA retrovirus expression vectors were reconstructed by molecular clone technology. Recombinant vectors were transfected into 293E package cells and virus suspension was collected. RT-PCR was used to detect the expression level of NF-κB P65 mRNA and TNF-α mRNA at different time-point of LPS stimulation. Western blot was performed to analyze the protein level of NF-κB P65. ELISA was applied to detect the expression level of TNF-α released by LPS-stimulated J774A.1. Results NF-κB P65 siRNA retrovirus expression vectors of mouse were successfully constructed. From2 hours after the stimulation of LPS, the expression level of NF-κB P65 mRNA of the siRNA group was obviously lower than the scramble control group ( 0.91 ±0.03 vs. 1.02 ±0.02, Plt;0.01) . At24,36, 48 and 72 hours after the LPS stimulation, the expression level of NF-κB P65 protein of the siRNA group was significantly decreased compared with the scramble control group ( 0.97 ±0.02 vs. 1.01 ±0.01, 0.94 ± 0.01 vs. 1.02 ±0. 01,0.94 ±0.02 vs. 1.02 ±0.01, 0.93 ±0.01 vs. 1.00 ±0.02, Plt;0. 05) . At 2, 6, 12, 24 hours after the LPS stimulation, both the expression level of TNF-α mRNA and the content of TNF-α in the culture medium supernatant of the siRNA group were lower than the scramble control group ( Plt;0. 01) . Conclusions The construction of NF-κB P65 siRNA retrovirus expression vectors is feasible. Inflammation factors in mouse monocyte-macrophages are significantly inhibited after NF-κB expression is depressed by RNA interference technology, which may be applied to prevent and treat excessive inflammatory reaction in acute lung injury.
ObjectiveTo enhance the understanding of Takayasu’s arteritis (TA) with pulmonary vascular involvement through the analyses on clinical features.MethodsA retrospective study was conducted to analyze the clinical records of patients diagnosed as TA with pulmonary vascular involvement admitted to Beijing Anzhen Hospital from January 2007 to August 2017.ResultsIn recent 10 years, there were 233 patients diagnosed as TA in Beijing Anzhen Hospital, 17 of them were involved with pulmonary arterial, which accounted for 7.3%. Fourteen patients were females, 3 patients were males. The age ranged from 16 to 59 years with an average of (40±13) years. Among the 17 patients, dyspnea (15 cases, 88.2%) was the most common symptom, and unequal blood pressure of upper limbs (9 cases, 52.9%) was the main sign. Thirteen patients had pulmonary hypertension by echocardiographic examination. Angiographic data showed that multi-lobular and multi-segmental pulmonary branches were predominantly affected. Unilateral involvement of pulmonary artery was more common than bilateral involvement, while the right pulmonary arteries were more often affected than the left. Thirteen patients received hormone and immunosuppressive therapy, 11 patients received anticoagulation or antiplatelet therapy, and 5 patients received targeted treatment of pulmonary artery pressure.ConclusionsThe clinical manifestation of TA is unspecific and misdiagnosis rate is relatively high. Improving the understanding of TA is necessary so as to reduce the misdiagnosis rate at an early stage and ameliorate the prognosis.
Objective To investigate the clinical characteristics and bacterial drug resistance of bloodstream infection of gram-negative bacteria, and provide guidance for clinical rational drug use and control of hospital infection. Methods A retrospective analysis was conducted in the patients diagnosed as severe pneumonia with blood culture of gram-negative bacteria from January 2015 to December 2017 in Beijing Anzhen Hospital. Results A total of 60 severe pneumonia patients suffered from bloodstream infection of gram-negative bacteria were recruited including 34 males and 26 females aging from 42 to 89 years and 73.4 years in average. In the 60 patients, 32 cases were infected with Klebsiella pneumonias, 20 cases were infected with Acinetobacter baumanni, and 8 cases were infected with Escherichia coli. The antimicrobial susceptibility testing result of Klebsiella pneumonias showed that the drug susceptibility rate was 100% to tigecycline, and 6.3% to amikacin. Escherichia coli was sensitive to Amikacin, imipenem, ceftazidime and meropenem while resistance to other drugs. The antimicrobial resistance of Acinetobacter baumanni was 28.6% for cefoperazone/sulbactam, and 14.3% for tigecycline. C-reactive protein, procalcitonin and SOFA scores were higher in the patients infected with Acinetobacter baumanni. Neutrophils and blood lactic acid were higher in the patients infected with Klebsiella pneumonias. There were no statistical differences in white blood cell, platelet or motality rate between the patients infected with Acinetobacter baumanni and the patients infected with Klebsiella pneumonias. SOFA scores and blood lactic acid had significantly statistical relevance with prognosis. Conclusion There is a high proportion of drug resistance of Klebsiella pneumoniae and Acinetobacter baumanni in the bloodstream infection of gram-negative bacteria.
ObjectiveTo study the correlation between international normalized ratio (INR) and coagulation factor Ⅱ and Ⅹ in patients with pulmonary thromboembolism treated with warfarin at moderate and low intensity anticoagulation.MethodsFifty-one patients with pulmonary thromboembolism treated with warfarin orally were divided into low-intensity anticoagulation group (INR from 1.6 to 2.0) and standard-intensity anticoagulation group (INR form 2.0 to 3.0) according to their monitoring INR indices. The levels of coagulation factor Ⅱ and Ⅹ were measured, and the correlation between INR level and coagulation factor activity was compared.ResultsThe INR of the low intensity anticoagulation group was 1.69±0.2 and the standard intensity anticoagulation group was 2.55±0.46. The corresponding activity of coagulation factor Ⅱ was (48.3±28.0)% and (24.0±8.0)% respectively. The activity of coagulation factor Ⅹ was (32.8±24.0)% and (16.7±6.0)%. There was a negative correlation between the activity of INR and coagulation factor Ⅱ and Ⅹ, with correlation coefficients of –0.903 and –0.459, respectively. Coagulation factor Ⅱ activity < 40%, coagulation factor Ⅹ activity inhibitory level < 25% is defined as anticoagulation effect. When coagulation factor Ⅱ activity level reaches anticoagulation effect, the corresponding minimum INR value was 1.56 and as to coagulation factor Ⅹ, the corresponding minimum INR value was 1.66.ConclusionsINR is negatively correlated with the activity of coagulation factor Ⅱ and coagulation factor Ⅹ. With the increase of INR, the activity of coagulation factor Ⅱ and coagulation factor Ⅹ decrease. Low intensity anticoagulation could not effectively inhibit the activity of coagulation factor.