1.Mechanisms of tigecycline resistance in carbapenems-resistant Acinetobacter baumannii strains
Meimei LI ; Jia DU ; Chunquan XU ; Huale CHEN ; Bin LI ; Yuanbo HOU ; Tieli ZHOU
Chinese Journal of Microbiology and Immunology 2015;35(10):717-722
Objective To investigate the mechanisms of tigecycline nonsusceptibility in carbapen-ems-resistant Acinetobacter baumannii ( CRAB) strains in order to provide a theoretical basis for a reasonable use of antibiotics and the control of nosocomial infection .Methods Susceptibility testing of 120 non-dupli-cate CRAB strains to tigecycline was performed by using the broth microdilution method .Minimal inhibitory concentrations ( MIC) of tigecycline against the A.baumannii strains were determined by using the broth mi-crodilution method before and after exposing the strains to Carbonylcyanide-m-chlorophenylhydrazone (CCCP), which was the efflux pump inhibitor .Polymerase chain reaction (PCR) was used to amply the ef-flux pumps genes including adeB, adeJ, adeG, abeM, adeE, adeRS, tetX and tetX1.The real-time PCR was performed to measure the expression of efflux pumps genes including adeB, adeJ, adeG, abeM and adeE.Results A total of 120 CRAB strains were collected including 13 (10.8%) tigecycline non-suscep-tible A.baumannii (TNAB) strains and 107 (89.2%) tigecycline susceptible A.baumannii (TSAB) strains.The MIC values of tigecline to the 120 CRAB strains were in a range of 0.25 μg/ml to 8 μg/ml. The adeR and adeJ genes were detected in 90.0%and 92.5%of the 120 CRAB strains, respectively.The positive rates of adeB, adeS, adeG and abeM genes among the 120 CRAB strains were all 94.2%.None of the three genes including adeE, tetX and tetX1 were detected .The mean expression levels of adeB and adeJ in TNAB strains were respectively increased by 18.69 folds and 5.46 folds as compared with those in sensi-tive strains.No significant increase in the expression of adeG and abeM genes was observed in TNAB strains . A 4-fold decrease in the MIC was observed in 8 out of 13 TNAB isolates treated with 10 μg/ml of CCCP .The CCCP could partially reverse the resistance pattern of tigecycline .Conclusion The efflux pump sys-tems of adeABC and adeIJK rather than the abeFGH and abeM systems might play an important role in reduc-ing the tigecycline susceptibility in carbapenems-resistant A.baumannii strains.
2.Study of mitochondrial energy metabolism in D-galactose-induced cell ageing model
Jianming CAO ; Dongjie HAO ; Huale CHEN ; Xiao YU ; Yuanbo HOU ; Wei YE
Chinese Journal of Geriatrics 2017;36(2):199-203
Objective To investigate the mitochondrial energy metabolism in D-galactoseinduced cell ageing model.Methods MRC-5 cells were cultivated for 72 hours in a medium containing 55 mmol/L D-galactose.The analysis of cell proliferation capacity by CCK8 method,β-galactosidase staining and detection of p21 protein expression level were performed for identifying cell senescence.The cell oxidation-reduction state was evaluated by an analysis of cellular ROS levels,SOD activity,MDA content and oxidative damage level of mitochondrial DNA(mtDNA).For purpose of detecting mitochondrial function and its impairment,mitochondrial morphology was observed by electron microscope,mitochondrial quantity was analyzed by flow cytometry,mitochondrial membrane potential(△Ψm) was measured by JC-1 staining,and ATP content was analyzed by HPLC,and mitochondrial oxygen consumption rate was detected by Seahorse cell energy metabolism detection system.Results The decreased MRC-5 cell proliferation,up-expression of p21 protein,increased β-galactosidase activity were observed in D-Gal-treated cells,which indicated the cell premature senescence.When treated with D-Gal,the significantly increased ROS and MDA level,decreased SOD activity and increased oxidized mtDNA proved that the cells kept higher oxidative stress.D-Gal induced-mitochondrial impairment was evidenced by the dimming of mitochondrial cristae and double membrane structure,decrease of transmembrane potential and ATP synthesis,and decrease of its oxygen consumption rate(OCR).Conclusions The 55 mmol/L D-Gal causes an impairment of mitochondrial structure and a decrease of function of energy metabolism,which is associated with cellular senescence induced by D-Gal.
4.Factors influencing success of external cephalic version and their clinical significance
Lianghui ZHENG ; Huale ZHANG ; Zhaodong LIU ; Qiuping LIAO ; Lichun CHEN ; Rongxin CHEN ; Jianying YAN
Chinese Journal of Perinatal Medicine 2023;26(1):11-19
Objective:To analyze the factors influencing the success rate of external cephalic version (ECV) and to create a preoperative scoring scale for stratified management of pregnant women who were preparing for ECV.Methods:This prospective study was conducted on singleton pregnant women who underwent ECV without anesthesia in Fujian Maternity and Child Health Hospital from January 1, 2017, to December 31, 2019. Univariate (two independent samples t-test, Mann-Whitney U test, and Chi-square test) and multivariate logistic regression were used to screen the clinical characteristics affecting the success of ECV, and receiver operating characteristic (ROC) curve was used to determine the cut-off value and convert quantitative variables into dichotomous variables. The independent variables were scored according to the regression coefficient in multivariate logistic regression analysis, and then a preoperative scoring scale was created. The ROC curve was used to calculate the cut-off value for the scoring scale. The subjects were divided into low and high score groups according to the cut-off value. The area under the ROC curve was used for evaluating the effectiveness of the scale in predicting the success of ECV. The success rate of ECV, difficulty of the operation and mode of delivery were compared between the two groups. Results:A total of 1 338 pregnant women met the inclusion criteria during the study period. After the exclusion of 885 women, 165 refused ECV in favor of direct cesarean section, 27 spontaneously converted to cephalic position before ECV, 261 who voluntarily accepted ECV were finally enrolled. ECV succeeded in 202 cases and failed in 59. (1) Favorable factors for ECV without anesthesia were the distance between the fetal breech and ischial spine <-3.5 cm ( OR=0.177, 95% CI: 0.071-0.438, P=0.009), the sum of the fundal height and the station of the fetal breech based on the ischial spine <30.25 cm ( OR=0.225, 95% CI: 0.094-0.537, P=0.001), amniotic fluid index ≥12 cm ( OR=0.399, 95% CI: 0.164-0.969, P=0.042), the surgeon's ability to hold the fetal head or breech with one hand ( OR=0.241, 95% CI: 0.098-0.589, P=0.002; OR=0.219, 95% CI: 0.087-0.546, P=0.001), and the fetal head located on the right or left upper abdomen of the mother ( OR=0.184, 95% CI: 0.059-0.568, P=0.003; OR=0.253, 95% CI: 0.084-0.760, P=0.014). (2) The area under the ROC curve of the preoperative score for predicting the success of ECV was 0.881 (95% CI: 0.821-0.941) and the cut-off value was 5.5. The subjects were divided into low (0-5 scores) and high (6-11 scores) score groups and the area under the ROC curve for predicting the success of ECV by grouping was 0.843 (95% CI: 0.774-0.912). Compared with the low score group, the high score group had a shorter ECV duration [2.0 min (0.5-10.0 min) vs 10.0 min (0.9-25.8 min), Z=-6.83, P<0.001], less attempts [1.0 times (1.0-4.0 times) vs 3.0 times (1.0-5.0 times), Z=-8.41, P<0.001], higher success rate [92.7% (190/205) vs 21.4% (12/56), χ2=127.64, P<0.001], higher rate of vaginal birth [75.4% (147/195) vs 18.5% (10/54)] and lower cesarean section rate [24.6% (48/195) vs 81.5% (44/54)] ( χ2=58.70, P<0.001). Conclusions:Preoperative scoring based on the factors influencing the success rate of ECV (the distance between the fetal breech and ischial spine, the sum of the fundal height and the station of the fetal breech based on the ischial spine <30.25 cm, amniotic fluid index ≥12 cm, the surgeon's ability to hold the fetal head or breech with one hand, and the fetal head locating on the right or left upper abdomen of the mother) is conducive to the individualized evaluation of the difficulty and the success rate of ECV as well as the success rate of vaginal delivery after ECV, which can provide a reference for clinical stratified management of ECV patients.