1.Constructed a cell line to express hBD1 stablly and detected the antimicrobial activity of hBD1 to multidrug resistant bacterial strains
Nan CUI ; Xinnian CHEN ; Lianhua WEI ; Juan LI ; Fengmei ZOU
Chinese Journal of Microbiology and Immunology 2011;31(12):1138-1142
ObjectiveTo established a cell line that expresses hBD1 stably,and detected the antimicrobial activity of the hBD1 to the muhidrug resistant bacterial strains.MethodsRecombinant plasmid was introduced into COS-7 cells by lipofectamine,cells were selected in culture medium containing G418 to acquired the monoclonal cell lines,total RNA were extracted from the cultured cells,expression levels of hBD1 mRNA was identified by RT-PCR,collected the supernatant solution of the cultured cell,expression levels of protein was identified by Western blot.Put the expression products and resistant organisms mixed together,after incubation in different times in 37℃,coating the mixtures in LB flat,then obtained the ratios between colonies number of experimental groups and colonies number of control groups,put those ratios as the survival rate of the drug resistance bacterias.Results The monoclonal cell lines had obtained after screened with G418,the hBD1 gene could be detected both at transcriptional and protein levels,Under the influence of expression product hBD1,survival rate of muhidrug-resistant Acinetobacter baumannii,multidrug-resistant Escherichia coli and multidrug-resistant Klebsiella pneumoniae could reduced to 9%,22% and 50%,but survival rate of multidrug-resistant Stenotrophomonas maltophilia is not have apparente difference with the control group.ConclusionThe stably-transfected cell line of hBD1 was successfully constructed,and the expression products of hBD1 showed the antimicrobial activity toward multidrug resistant bacterial strains.
2.Correlation Study of Cognitive Disorder Induced by Ethanol and M1 Receptor in Hippocampus in Mice
Dan ZOU ; Minfan WU ; Ge JIN ; Yan ZHENG ; Fengmei CHEN
Journal of China Medical University 2015;(7):602-605
Objective To investigate the effect of ethanol on level of the main hippocampal subtype of muscarinic receptor(M1)in mice,and evalu?ate whether the content change on this receptor could be linked with alterations in cognition,so as to further reveal the mechanism of brain damage in?duced by ethanol. Methods Sixty female mice were randomly divided into four groups. The model mice were induced by intragastric administration of ethanol at dose of 8%,16%,and 32%respectively of 0.2 mL/10 g for 8 weeks according to the protocol,and control group were treated with intra?gastric administration of distilled water. The capability of learning and memory were examined by Morris water maze,and ELISA method was used to measure the M1 receptor content in hippocampus in each group of mice. Results Compared with first day,the mean escape latency period on the fifth day was significantly shortened in each group. There was no significant difference between ethanol and control group for the mean escape latency period on the fifth day. Compared with the control group,the active time in the target quadrant was significantly shortened in 16%and 32%ethanol group. M1 receptor content in hippocampus formation was significantly decreased in all the ethanol group mice. The ethanol concentration was nega?tive correlated with the M1 receptor content. Conclusion Chronic alcoholism can induce the memory impairment in mice,which might be associat?ed with the low level of M1 receptor subtype in hippocampus of mice.
3.Establishment and application of external quality assessment system for microbial morphology and detection of special drug-resistance in clinical laboratory
Lianhua WEI ; Fengmei ZOU ; Gang LIU ; Yongqing YANG ; Wanxia WANG ; Ling WU ; Junchun LI ; Xin WANG ; Yuqian CHEN ; Xiaoqing CHEN ; Keke LI ; Haixing SHAO ; Jichao ZHU
Chinese Journal of Clinical Laboratory Science 2017;35(7):538-541
Objective To investigate the establishment,operation and performance of external quality assessment(EQA) system for microbial morphology and detection of special drug-resistance in clinical laboratory,and explore the value of the developed system in clinical application.Methods The pictures of known bacteria and fungi colony,gram staining and acid-fast staining from clinical microbiology were distributed to the participating laboratories in Gansu province twice a year at regular intervals.The pictures of standard knowledge points from CLSI,such as special drug resistance were distributed simultaneously.All the participating laboratories were required to complete the interpretation for the pictures and report their resuhs in a scheduled time.Then the resuhs were summarized and analyzed as 3 modes:complete consistency,general consistency and non-consistency.Results During the 2 years when the EQA system for microbial morphology and detection of special drug-resistance were performed for 24 times,the rate of annual complete consistency increased year by year and reached to 91.3% in 2015.Conclusion The EQA system based on the examinations of microbial morphology and CLSI standard knowledge points for clinical laboratory may supervise the staff of clinical microbiology laboratories in the hospitals at second grade or above to master the skills of morphological identification and learn CLSI knowledge points,so their professional skills of clinical microbiology could be comprehensively improved.
4.Antimicrobial resistance profile of clinical isolates in hospitals across China: report from the CHINET Surveillance Program, 2017
Fupin HU ; Yan GUO ; Demei ZHU ; Fu WANG ; Xiaofei JIANG ; Yingchun XU ; Xiaojiang ZHANG ; Zhaoxia ZHANG ; Ping JI ; Yi XIE ; Mei KANG ; Chuanqing WANG ; Aimin WANG ; Yuanhong XU ; Jilu SHEN ; Ziyong SUN ; Zhongju CHEN ; Yuxing NI ; Jingyong SUN ; Yunzhuo CHU ; Sufei TIAN ; Zhidong HU ; Jin LI ; Yunsong YU ; Jie LIN ; Bin SHAN ; Yan DU ; Sufang GUO ; Lianhua WEI ; Fengmei ZOU ; Hong ZHANG ; Chun WANG ; Yunjian HU ; Xiaoman AI ; Chao ZHUO ; Danhong SU ; Ruizhong WANG ; Hua FANG ; Bixia YU ; Yong ZHAO ; Ping GONG ; Dawen GUO ; Jinying ZHAO ; Wenen LIU ; Yanming LI ; Yan JIN ; Chunhong SHAO ; Kaizhen WEN ; Yirong ZHANG ; Xuesong XU ; Chao YAN ; Hua YU ; Xiangning HUANG ; Shanmei WANG ; Yafei CHU ; Lixia ZHANG ; Juan MA ; Shuping ZHOU ; Yan ZHOU ; Lei ZHU ; Jinhua MENG ; Fang DONG ; Hongyan ZHENG ; Han SHEN ; Wanqing ZHOU ; Wei JIA ; Gang LI ; Jinsong WU ; Yuemei LU
Chinese Journal of Infection and Chemotherapy 2018;18(3):241-251
Objective To investigate the antimicrobial resistance profile of the clinical isolates collected from selected hospitals across China. Methods Twenty-nine general hospitals and five children's hospitals were involved in this program. Antimicrobial susceptibility testing was carried out according to a unified protocol using Kirby-Bauer method or automated systems. Results were interpreted according to CLSI 2017 breakpoints. Results A total of 190 610 clinical isolates were collected from January to December 2017, of which gram negative organisms accounted for 70.8% (134 951/190 610) and gram positive cocci 29.2% (55 649/190 610). The prevalence of methicillin-resistant strains was 35.3% in S. aureus (MRSA) and 80.3% in coagulase negative Staphylococcus (MRCNS) on average. MR strains showed much higher resistance rates to most of the other antimicrobial agents than MS strains. However, 91.6% of MRSA strains were still susceptible to trimethoprim-sulfamethoxazole, while 86.2% of MRCNS strains were susceptible to rifampin. No staphylococcal strains were found resistant to vancomycin. E. faecalis strains showed much lower resistance rates to most of the drugs tested (except chloramphenicol) than E. faecium. Vancomycin-resistant Enterococcus (VRE) was identified in both E. faecalis and E. faecium. The identified VRE strains were mainly vanA, vanB or vanM type based on phenotype or genotype. The proportion of PSSP or PRSP strains in the non-meningitis S.pneumoniae strains isolated from children decreased but the proportion of PISP strains increased when compared to the data of 2016. Enterobacteriaceae strains were still highly susceptible to carbapenems. Overall, less than 10% of these strains (excluding Klebsiella spp.) were resistant to carbapenems. The prevalence of imipenem-resistant K. pneumoniae increased from 3.0% in 2005 to 20.9% in 2017, and meropenem-resistant K. pneumoniae increased from 2.9% in 2005 to 24.0% in 2017, more than 8-fold increase. About 66.7% and 69.3% of Acinetobacter (A. baumannii accounts for 91.5%) strains were resistant to imipenem and meropenem, respectively. Compared with the data of year 2016, P. aeruginosa strains showed decreasing resistance rate to carbapenems. Conclusions Bacterial resistance is still on the rise. It is necessary to strengthen hospital infection control and stewardship of antimicrobial agents. The communication between laboratorians and clinicians should be further improved in addition to surveillance of bacterial resistance.