| Objective: PComplex Intra-abdominal Infections(c IAI)is one of the more common infections in clinic.CIAI generally occurs in the abdominal cavity,behind the abdominal organs and peritoneum.In most cases,c IAI is a mixed bacterial infection.The infection sources include Gram-negative anaerobes,Aerobic Bacilli,positive aerobic and anaerobic bacteria,etc.,and broad-spectrum antibiotics or combinations of antibiotics are required.Tigecycline is a derivative of minocycline,tigecycline broad antibacterial spectrum,tigecycline for the treatment of c IAI positive effect,for the clinical first-line drugs.This study collected a total of 24 cases of c IAI patients from department of ICU of the Second Affiliated Hospital of Anhui Medical University during June 2015 to June 2017,and the other 15 cases of healthy volunteers from medical examination center were also enrolled.To observe the therapeutic effect of tigecycline on patients with c IAI,in the observation of tigecycline c IAI patients play an anti-infective role at the same time.To further study the effect of tigecycline on immune function in patients with c IAI in vitro and in vivo,and to provide some experimental evidence for further study on the anti-infective effect of tigecycline.Objective:To study the effect of tigecycline on immune function in patients with complicated intra-abdominal infection.Method:1.The diagnostic criteria of 1.cIAIWith c IAI surgical indications(laparotomy,laparoscopic surgery or percutaneous drainage abscess surgery),secondary surgery or imaging studies.The exclusion criteria were as follows: abdominal abscess,small bowel obstruction,and female genital tract infection.Other systemic antibiotics were included within 72 hours prior to the study,with low blood volume(<25%)and platelet count less than 75 000 / mm \ or neutrophil count less than 1000 / mm \ as well as other life-threatening illnesses(acute liver failure and liver disease)and patients with a history of antibiotic use due to intraperitoneal infection.2.Treatment options and efficacy evaluationCIAI patients treatment options are as follows,tigecycline(Pfizer Pharmaceuticals Tiger,batch number: H20143394)first dose of 100 mg,maintenance dose 50 mg / 12 h,intravenous infusion,the treatment of 7 to 14 days,the clinical symptoms disappeared,laboratory tests and bacteriological examination results;the above results were negative for significant and effective;after treatment,patients with clinical symptoms and signs disappeared or significantly improved,and laboratory tests and bacteriological results were recorded as part of the improvement;treatment clinical symptoms before and after the patient,laboratory tests and bacteriological findings no significant change as invalid.VITEK-2 automatic microbiological analysis system(France Merial biotechnology company)to identify the pathogen isolated from abdominal infection specimens.3.PBMC preparation and proliferationPeripheral blood mononuclear cells(PBMC)were prepared from c IAI patients and healthy volunteers by gradient centrifugation before and after the intervention of tigecycline.The above cells were seeded in 96-well plates at 1 × 10 \ per well in a final volume of 200 ?l per well.The experimental groups were as follows: normal group,model group,1 mg / L,2 mg / L and 5 mg / L tigecycline group,except the normal group,each group were treated with 100 ?g / L Staphylococcus aureus Toxin A stimulated for 24 h.Four hours before the end of the experiment,100 ?l MTT solution(5 g / L)was added and the culture was continued for 4 h.The culture was stopped and the absorbance value(A)of each well was detected by microplate reader(Bio Tek USA).4.Cytokines level detectionTake healthy volunteers,before treatment and after treatment of patients with c IAI 3 ml of peripheral blood,serum was separated.PBMC cells were prepared as before.The levels of interleukin-1(IL-1),IL-6 and IL-8 were measured according to the instructions of RD Systems of USA.5.Procalcitonin,C-reactive protein,blood testTake healthy volunteers,before treatment and after treatment of patients with c IAI 5 ml of peripheral blood,serum was separated.The procalcitonin level was measured in a Modular PPI automatic biochemical analyzer according to the procedure for the quantitative detection of the kit by procalcitonin electrochemiluminescence.C-reactive protein levels were measured on a Modular PPI automated biochemical analyzer following C-reactive protein diagnostic kit protocol.Peripheral blood leukocyte and neutrophil levels were measured according to the XE5000 automated hemacytometer procedures.6.Detection of 6.PBMC T cell subsets levelBefore and after treatment,healthy volunteers were drawn,before treatment and after treatment of patients with c IAI peripheral blood 2 ml,adding anticoagulant EDTA.Take100 ?l of anticoagulant into the tube and add 10 ?l of fluorescent labeled antibody for staining and mixing,including CD3 \,CD4 \ and CD8 \ antibody(Santa Cruz,USA).Incubate at room temperature for 30 minutes in the dark,add 500 ?l of hemolysin to lyse erythrocytes,and incubate at room temperature for 20 minutes in the dark to detect by flow cytometry.7.Statistical AnalysisUse SPSS16.0 software for analysis,count data as a percentage,measurement data to Said that the same group of sample data before and after treatment compared with the level of immunity paired sample t test,the immune level between the two groups compared with independent samples t test,P <0.05 said the difference was statistically significant.Result:1.Tigecycline effectively inhibits c IAI patients with intra-abdominal infectionThe total effective rate of tigecycline in patients with c IAI was 70.8%(effective rate was 45.80% + effective rate was 25.0%),the inefficiency was 29.2%.The inhibitory rates of tigecycline against pathogenic microorganisms were 70.6%(E.coli),42.9%(Enterobacter cloacae),55.6%(Acinetobacter baumannii),75.0%(Clostridium),72.7%(Digestive chain77.8% of enterococci,60.0% of Staphylococcus aureus,72.7% of S.anguillarum,55.6% of Proteus mirabilis,83.3% of Klebsiella pneumoniae.2.Effect of tigecycline on peripheral blood leukocytes and neutrophils in c IAI patientsCIAI patients in the use of tigecycline before treatment,peripheral blood leukocyte and neutrophil levels increased significantly compared with the normal group,there is significant difference \ of P<0.01,c IAI in patients treated by using the add ring,peripheral blood leukocyte and neutrophils the level was significantly decreased compared with before treatment,there is significant difference \ P<0.01).3.Effect of tigecycline on c-reactive protein and procalcitonin in patients with c IAICIAI patients in the use of tigecycline before treatment,C reactive protein and calcitonin in peripheral blood were significantly increased compared with the normal group,there is significant difference \ of P<0.01,c IAI in patients treated by using the add ring,C reactive protein and calcitonin in peripheral blood the fibrinogen level compared with before treatment was significantly reduced,there is significant difference \ P<0.01).4.Tigecycline reduces the levels of inflammatory cytokines in serum and PBMC of patients with c IAICompared with the normal group,the levels of serum inflammatory cytokines IL-1?,IL-6and IL-8 in c IAI patients before treatment with tigecycline were significantly higher(P<0.01);The levels of IL-1?,IL-6 and IL-8 in c IAI patients after treatment with tigecycline were significantly lower than those before treatment,the difference was statistically significant \ P <0.01.In addition,PBMCs were prepared in vitro and the difference was statistically significant(P <0.01)when SEA stimulation was used to significantly increase the levels of IL-1?,IL-6 and IL-8 secreted by cells Drugs could significantly reduce the level of IL-1?,IL-6 and IL-8 in PBMC supernatant,the difference was statistically significant \ P <0.01,suggesting that tigecycline directly reduce the inflammatory cytokines Level.5.Tigecycline reduces PBMC proliferation in c IAI patientsThe results showed that compared with the PBMC of normal control group,SEA stimulation in vitro could significantly increase the proliferation of PBMC,the difference was statistically significant \ P <0.01.In vitro administration of tigecycline significantly reduced the proliferation of PBMCs stimulated by SEA,the difference was statistically significant(P <0.01).The results suggest that tigecycline has a direct inhibitory effect on the proliferation of PBMCs stimulated by SEA.6.Tigecycline regulates the expression of CD3,CD4 and CD8 in peripheral blood of c IAI patientsCompared with the normal control group,the percentages of CD48 \ and CD4 \ in peripheral blood of patients with c IAI before treatment were significantly lower,the ratios of CD4 \ / CD8 \ and the percentage of CD8 The difference was statistically significant(P<0.01).Compared with those before c IAI treatment,the ratio of TF53,CD4 \ and CD4 \ /CD8 \ in peripheral blood were significantly increased after treatment with tigecycline While reducing the percentage of CD8 \,the difference was statistically significant P <0.01,suggesting that tigecycline in c IAI patients in vivo immune imbalance there is a certain regulatory role.In conclusion:1.Tigecycline has a therapeutic effect on c IAI patients.2.Tigecycline in the play This study shows that tigecycline in the role of anti-infective at the same time,in vitro and in vivo effects of c IAI immune cells,can regulate the body immune imbalance. |
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