Egg-associated secretions from the brown planthopper (Nilaparvata lugens) activate rice immune responses.

The brown planthopper (BPH, Nilaparvata lugens) is a notorious sap-sucking insect pest that damages rice (Oryza sativa) plants throughout Asia. During BPH feeding, saliva enters rice plant tissues, whereas during oviposition egg-associated secretions (EAS) are deposited in damaged plant tissue. Dynamic changes in rice to planthopper salivary effectors have been widely reported. However, the effects of EAS from planthopper on rice immunity remains largely unexplored. In this study, we found that both infestation of rice by gravid BPH female adults and treatment with the EAS elicited a strong and rapid accumulation of jasmonic acid (JA), JA-isoleucine, and hydrogen peroxide in rice. EAS enhanced plant defenses not only in rice but also in tobacco, and these impaired the performance of BPH on rice, as well as the performance of aphids and whiteflies on tobacco. High-throughput proteome sequencing of EAS led to 110 proteins being identified and 53 proteins with 2 or more unique peptides being detected. Some proteins from BPH EAS were also found in the salivary proteome from herbivores, suggesting potential evolutionary conservation of effector functions across feeding and oviposition; however, others were only identified in EAS, and these are likely specifically related to oviposition. These findings point to novel proteins affecting interactions between planthoppers and rice during oviposition, providing an additional source of information for effector studies.

Combined transcriptome and metabolome analysis reveal that the white and yellow mango pulp colors are associated with carotenoid and flavonoid accumulation, and phytohormone signaling.

Mango (Mangifera indica L.) is a widely appreciated tropical fruit for its rich color and nutrition. However, knowledge on the molecular basis of color variation is limited. Here, we studied HY3 (yellowish-white pulp) and YX4 (yellow pulp), reaped with 24 h gap from the standard harvesting time. The carotenoids and total flavonoids increased with the advance of harvest time (YX4 > HY34). Transcriptome sequencing showed that higher expressions of the core carotenoid biosynthesis genes and flavonoid biosynthesis genes are correlated to their respective contents. The endogenous indole-3-acetic acid and jasmonic acid contents decreased but abscisic acid and ethylene contents increased with an increase in harvesting time (YX4 > HY34). Similar trends were observed for the corresponding genes. Our results indicate that the color differences are related to carotenoid and flavonoid contents, which in turn are influenced by phytohormone accumulation and signaling.

Phylogenetic Analysis Reveals Distinct Evolutionary Trajectories of the Fluoroquinolones-Resistant Escherichia coli ST1193 From Fuzhou, China.

Escherichia coli (E. coli) ST1193 is an emerging fluoroquinolones-resistant and virulent lineage. Large gaps remain in our understanding of the evolutionary processes and differences of this lineage. Therefore, we used 76 E. coli ST1193 genomes to detect strain-level genetic diversity and phylogeny of this lineage globally. All E. coli ST1193 possessed fimH64, filCH5, and fumC14. There was 94.7% of isolates classified as O-type O75. There was 9.33% of E. coli ST1193 that possessed K5 capsular, while 90.67% of isolates possessed K1 capsular. The core genome analysis revealed that all isolates were divided into two phylogenetic clades (clade A and B). Clade A included 25 non-Chinese E. coli ST1193, and clade B contained all isolates collected from Fuzhou, China, respectively. The results of comparative genomics indicated Indels were identified in 150 clade-specific genes, which were enriched into the biological process and molecular function. Accessory genome phylogenetic tree showed a high degree of correlation between accessory genome clusters and core genome clades. There was significant difference in antibiotic resistance genes (ARGs) [bla CTX-M-55 , bla TEM-1 , sul2, tet(B), tet(R), APH(6)-Id, and AAC(3)-IId], virulence factors (cia, neuC, gad, and traT), and plasmid replicon types (IncQ1, Col156, and IncB/O/K/Z) between clade A (non-Chinese isolates) and clade B (Chinese isolates) (p < 0.05). Further analysis of the genetic environments of bla CTX-M-55 demonstrated that the flanking contexts of bla CTX-M-55 were diverse. In conclusion, our results reveal the distinct evolutionary trajectories of the spread of E. coli ST1193 in Fuzhou, China and non-China regions. This supports both global transmission and localized lineage expansion of this lineage following specific introductions into a geographic locality.

Synthesis and biological evaluation of N 6 derivatives of 8-azapurine as novel antiplatelet agents.

Two series of novel N 6 derivatives of 8-azapurine I and II were designed as antiplatelet agents. Series I and II were N 6 amino derivatives and N 6 hydrazone derivatives of 8-azapurine, respectively. The compounds were synthesized in acceptable yields via conventional procedures, including nucleophilic substitution, diazotization, and amination or hydrazonation with amino alcohol and 4,6-dichloropyrimidine as starting materials. To assess the ability of the synthesized compounds as antiplatelet agents, the ADP-induced platelet aggregation assay of Born was performed both in vitro and in vivo using ticagrelor as a reference control substance. The analysis of the structure-activity relationship and molecular docking were also discussed in detail. The results demonstrated that series I and II compounds exhibited antiplatelet activity in vitro and IIh was the most active compound (IC50 = 0.20 µM) among the target compounds, being almost 4-fold better than ticagrelor (IC50 = 0.74 µM). For a preliminary assessment of the safety profile, a bleeding test (mouse tail) and a single-dose toxicity test were conducted. The use of compound IIh resulted in a shorter bleeding time, less blood loss and lower acute toxicity compared to ticagrelor. In addition, a molecular docking study was performed to investigate the binding capacity and binding mode between IIh and P2Y12.

Synthesis and biological evaluation of N4 -hydrazone derivatives of 5,7-dihydro-6H-pyrrolo[2,3-d]pyrimidin-6-one as novel anticancer agents with antimetastatic adjunct efficacy.

To obtain new anticancer agents with antimetastatic adjunct efficacy, a series of novel N4 -hydrazone derivatives of 5,7-dihydro-6H-pyrrolo[2,3-d]pyrimidin-6-one were designed and synthesized by an eight-step reaction, with appropriate yields. All the synthesized compounds were evaluated for their antiproliferative activity against A549 and MCF-7 cells and for antiplatelet aggregation activity in vitro. The results showed that compounds 25 and 35 not only showed potent antiproliferative activity against the A549 (IC50 = 15.3 and 21.4 µM) and MCF-7 (IC50 = 15.6 and 10.9 µM) cell lines but also showed certain antiplatelet aggregation activity (inhibition rates: 47.0% and 45.8%). These results indicated that the structural modification on the N4 -hydrazone moiety of 5,7-dihydro-6H-pyrrolo[2,3-d]pyrimidin-6-one is promising to obtain novel anticancer compounds with antimetastatic adjunct efficacy. In addition, a molecular docking study was performed to investigate the possible targets, and these results indicated that compounds 25 and 35 have the potential to target EGFR, HER2, and P2Y12 .

Acquisition of a Stable and Transferable blaNDM-5-Positive Plasmid With Low Fitness Cost Leading to Ceftazidime/Avibactam Resistance in KPC-2-Producing Klebsiella pneumoniae During Treatment.

The emergence and prevalence of carbapenem-resistant Enterobacteriaceae (CRE) have drawn worldwide attention. Ceftazidime/avibactam (CAZ/AVI) gives us a valuable alternative strategy to treat CRE infections. Unfortunately, CAZ/AVI resistance could occur during CAZ/AVI treatment. The CAZ/AVI-resistant Carbapenem-resistant Klebsiella pneumoniae (CR-KP) (KP137060) and earlier CAZ/AVI-susceptible isolate (KP135194) from the same hospitalized patient were collected at Fujian Medical University Union Hospital between October and November 2019. In this study, CAZ/AVI MICs of CAZ/AVI-susceptible and -resistant isolates (KP135194 and KP137060) were 4 mg/L and 128 mg/L, respectively; and the two isolates had the same antibiotic resistance pattern to other carbapenems. Two strains were then submitted for whole-genome sequencing and bioinformatic analysis. ompK36 was not detected in two isolates. No mutation was observed in blaKPC-2, ompK35 and ompK37 in this study and there was no significant difference of the expression in blaKPC-2, ompK35 and ompK37 between the two isolates (p>0.05). Two isolates were sequence type 11 and harbored blaKPC-2, blaSHV-182 and blaTEM-1B. Compared with KP135194, KP137060 harbored an additional blaNDM-5 positive plasmid. blaNDM-5 gene could be successfully transferred into E. coli J53 at a conjugation frequency of 1.14×10-4. Plasmid stability testing showed that blaKPC-2- and blaNDM-5-harboring plasmids were still stably maintained in the hosts. Growth assay and growth competition experiments showed there was no significant difference in fitness cost between two CR-KP isolates. Our study described the acquisition of a blaNDM-5-harboring plasmid leading to resistance to ceftazidime/avibactam in KPC-2-producing Klebsiella pneumoniae during treatment. This phenomenon deserves further exploration.

Epidemic Potential of Escherichia coli O16:H41-ST131: Compared with Pandemic O25b:H30-ST131 Lineage.

BACKGROUND: O16:H41 is an important subclone among Escherichia coli (E. coli) sequence type (ST) 131, which has risen dramatically in recent years. However, reasons for the rapid increase of E. coli O16:H41-ST131 remain unclear. The aim of this study was to compare the pathogenicity and survivability features of E. coli O16:H41-ST131 with global epidemic O25b:H30-ST131 lineage. METHODS: Sixteen E. coli ST131 were divided into two groups: group O16:H41-ST131 (n=6) and group O25b:H30-ST131 (n=10). Adhesion and invasion activity of different isolates were measured using human T24 cells. Biofilm production was quantified by crystal violet staining. Fifty percent human serum was used to detect serum sensitivity. Resistance to hydrogen peroxide was detected by broth microdilution method, and anti-phagocytic function was determined by phagocytosis experiments. RESULTS: E. coli O16:H41-ST131 and O25b:H30-ST131 lineage showed similar biofilm formation, adhesion and invasion abilities. In terms of survivability, resistance to serum and hydrogen peroxide of E. coli O16:H41-ST131 was similar as that of E. coli O25b:H30-ST131. But anti-phagocytic function of E. coli O16:H41-ST131 was significantly weaker than that of E. coli O25b:H30-ST131. CONCLUSION: The pathogenicity and survivability of E. coli O16:H41-ST131 were similar to those of E. coli O25b:H30-ST131, which may be important reasons for its increasing prevalence. Our study may contribute to a better understanding of the prevalence of E. coli O16:H41-ST131.

A network pharmacology-based investigation on the bioactive ingredients and molecular mechanisms of Gelsemium elegans Benth against colorectal cancer.

BACKGROUND: Colorectal cancer (CRC) remains one of the leading causes of cancer-related death worldwide. Gelsemium elegans Benth (GEB) is a traditional Chinese medicine commonly used for treatment for gastrointestinal cancer, including CRC. However, the underlying active ingredients and mechanism remain unknown. This study aims to explore the active components and the functional mechanisms of GEB in treating CRC by network pharmacology-based approaches. METHODS: Candidate compounds of GEB were collected from the Traditional Chinese Medicine@Taiwan, Traditional Chinese Medicines Integrated Database, Bioinformatics Analysis Tool for Molecular mechanism of Traditional Chinese Medicine, and published literature. Potentially active targets of compounds in GEB were retrieved from SwissTargetPrediction databases. Keywords "colorectal cancer", "rectal cancer" and "colon cancer" were used as keywords to search for related targets of CRC from the GeneCards database, then the overlapped targets of compounds and CRC were further intersected with CRC related genes from the TCGA database. The Cytoscape was applied to construct a graph of visualized compound-target and pathway networks. Protein-protein interaction networks were constructed by using STRING database. The DAVID tool was applied to carry out Gene Ontology and Kyoto Encyclopedia of Genes and Genome pathway enrichment analysis of final targets. Molecular docking was employed to validate the interaction between compounds and targets. AutoDockTools was used to construct docking grid box for each target. Docking and molecular dynamics simulation were performed by Autodock Vina and Gromacs software, respectively. RESULTS: Fifty-three bioactive compounds were successfully identified, corresponding to 136 targets that were screened out for the treatment of CRC. Functional enrichment analysis suggested that GEB exerted its pharmacological effects against CRC via modulating multiple pathways, such as pathways in cancer, cell cycle, and colorectal cancer. Molecular docking analysis showed that the representative compounds had good affinity with the key targets. Molecular dynamics simulation indicated that the best hit molecules formed a stable protein-ligand complex. CONCLUSION: This network pharmacology study revealed the multiple ingredients, targets, and pathways synergistically involved in the anti-CRC effect of GEB, which will enhance our understanding of the potential molecular mechanism of GEB in treatment for CRC and lay a foundation for further experimental research.

Molecular Mechanism of Gelsemium elegans (Gardner and Champ.) Benth. Against Neuropathic Pain Based on Network Pharmacology and Experimental Evidence.

Gelsemium elegans (Gardner and Champ.) Benth. (Gelsemiaceae) (GEB) is a toxic plant indigenous to Southeast Asia especially China, and has long been used as Chinese folk medicine for the treatment of various types of pain, including neuropathic pain (NPP). Nevertheless, limited data are available on the understanding of the interactions between ingredients-targets-pathways. The present study integrated network pharmacology and experimental evidence to decipher molecular mechanisms of GEB against NPP. The candidate ingredients of GEB were collected from the published literature and online databases. Potentially active targets of GEB were predicted using the SwissTargetPrediction database. NPP-associated targets were retrieved from GeneCards, Therapeutic Target database, and DrugBank. Then the protein-protein interaction network was constructed. The DAVID database was applied to Gene Ontology and Kyoto Encyclopedia of Genes and Genome pathway enrichment analysis. Molecular docking was employed to validate the interaction between ingredients and targets. Subsequently, a 50 ns molecular dynamics simulation was performed to analyze the conformational stability of the protein-ligand complex. Furthermore, the potential anti-NPP mechanisms of GEB were evaluated in the rat chronic constriction injury model. A total of 47 alkaloids and 52 core targets were successfully identified for GEB in the treatment of NPP. Functional enrichment analysis showed that GEB was mainly involved in phosphorylation reactions and nitric oxide synthesis processes. It also participated in 73 pathways in the pathogenesis of NPP, including the neuroactive ligand-receptor interaction signaling pathway, calcium signaling pathway, and MAPK signaling pathway. Interestingly, 11-Hydroxyrankinidin well matched the active pockets of crucial targets, such as EGFR, JAK1, and AKT1. The 11-hydroxyrankinidin-EGFR complex was stable throughout the entire molecular dynamics simulation. Besides, the expression of EGFR and JAK1 could be regulated by koumine to achieve the anti-NPP action. These findings revealed the complex network relationship of GEB in the "multi-ingredient, multi-target, multi-pathway" mode, and explained the synergistic regulatory effect of each complex ingredient of GEB based on the holistic view of traditional Chinese medicine. The present study would provide a scientific approach and strategy for further studies of GEB in the treatment of NPP in the future.

Hospital Wastewater as a Reservoir for Antibiotic Resistance Genes: A Meta-Analysis.

Background: The emergence and dissemination of antibiotic resistance genes (ARGs) in the environment poses a huge global health hazard. Hospital wastewater (HWW), in which a high density of antibiotic residues and antibiotic-resistant bacteria are present, may be a reservoir of ARGs dissemination into the environment. Our meta-analysis comprehensively analyzes the prevalence of ARGs in HWW, as well as the influencing factors in ARGs distribution. Methods: Online databases were used to search for literature using the subject terms: "Drug Resistance" AND "Genes" AND "Hospitals" AND "Wastewater." Two reviewers independently applied predefined criteria to assess the literature and extract data including "relative abundance of ARGs," "title," "authors," "country," "location," "sampling year," and "sampling seasons." The median values and 95% confidence intervals of ARGs abundance were calculated by Wilcox.test function in R. Temporal trends, spatial differences, seasonal variations and removal efficiency of ARGs were analyzed by Pearson correlation analysis and Kruskal-Wallis H test. Results: Resistance genes to carbapenems, sulfonamides, tetracyclines and mobile genetic elements were found at high relative abundance (>10-4 gene copies/16S rRNA gene copies) in HWW. The abundance of resistance genes to extended-spectrum ß-lactams, carbapenems, sulfonamides and glycopeptide significantly decreased, while tetracycline resistance genes abundance increased from 2014 to 2018. The abundance of ARGs was significantly different by country but not by season. ARGs could not be completely removed by on-site HWW treatments and the removal efficiency varies for different ARGs. Conclusions: HWW presents more types of ARGs, and their abundance is higher than those in most wastewater systems. HWW may be a reservoir of ARGs and play an important role in the dissemination of ARGs.

A Comparative Study of Fluoroquinolone-Resistant Escherichia coli Lineages Portrays Indistinguishable Pathogenicity- and Survivability-Associated Phenotypic Characteristics Between ST1193 and ST131.

BACKGROUND: Sequence type 1193 is a new such lineage among fluoroquinolone-resistant Escherichia coli, which has risen dramatically within the last several years. However, reasons for rapid emergence and successful spread of E. coli ST1193 remain unclear. The aim of this study was to compare the pathogenicity and survivability features of E. coli ST1193 with global epidemic lineage, ST131. METHODS: A total of 30 E. coli were used in this study. Isolates were divided into two groups, ST1193 (n=15) and ST131 (n=15). Adhesion and invasion to T24 cells and resistance to serum were quantified and compared among two groups. Biofilm formation capacity was assessed by crystal violet assay. Macrocolony formation was assessed on macrocolony formation plates. Resistance to hydrogen peroxide was performed by broth microdilution. RAW264.7 cells were used to assess the anti-phagocytic function of different isolates. RESULTS: Adhesion and invasion assays revealed that E. coli ST1193 could adhere and invade T24 cells (p <0.05). 93.3% of E. coli ST1193 could form biofilms. The majority of E. coli ST1193 (66.7%) possessed no curli/no cellulose on macrocolony formation plates. E. coli ST1193 showed significant growth in serum and hydrogen peroxide and illustrated higher anti-phagocytic function to RAW264.7 cells (p <0.05). Group analysis showed that E. coli ST1193 was similar to ST131 in pathogenicity- and survivability-associated phenotypic characteristics (p >0.05). CONCLUSION: Our study provided more insights into pathogenicity and survivability features of E. coli ST1193, which was similar to ST131. Our study could be of great importance in understanding the emergence of global spread E. coli ST1193. Strategic and continued surveillance should be carried out to prevent the infections caused by E. coli ST1193.

The genome evolution and domestication of tropical fruit mango.

BACKGROUND: Mango is one of the world's most important tropical fruits. It belongs to the family Anacardiaceae, which includes several other economically important species, notably cashew, sumac and pistachio from other genera. Many species in this family produce family-specific urushiols and related phenols, which can induce contact dermatitis. RESULTS: We generate a chromosome-scale genome assembly of mango, providing a reference genome for the Anacardiaceae family. Our results indicate the occurrence of a recent whole-genome duplication (WGD) event in mango. Duplicated genes preferentially retained include photosynthetic, photorespiration, and lipid metabolic genes that may have provided adaptive advantages to sharp historical decreases in atmospheric carbon dioxide and global temperatures. A notable example of an extended gene family is the chalcone synthase (CHS) family of genes, and particular genes in this family show universally higher expression in peels than in flesh, likely for the biosynthesis of urushiols and related phenols. Genome resequencing reveals two distinct groups of mango varieties, with commercial varieties clustered with India germplasms and demonstrating allelic admixture, and indigenous varieties from Southeast Asia in the second group. Landraces indigenous in China formed distinct clades, and some showed admixture in genomes. CONCLUSIONS: Analysis of chromosome-scale mango genome sequences reveals photosynthesis and lipid metabolism are preferentially retained after a recent WGD event, and expansion of CHS genes is likely associated with urushiol biosynthesis in mango. Genome resequencing clarifies two groups of mango varieties, discovers allelic admixture in commercial varieties, and shows distinct genetic background of landraces.

Discovery and optimization of 5,7-dihydro-6H-pyrrolo[2,3-d]pyrimidin-6-one derivatives as mTORC1/mTORC2 dual inhibitors.

New potent mTORC1/mTORC2 dual inhibitors, 5,7-dihydro-6H-pyrrolo[2,3-d]pyrimidin-6-one derivatives, were obtained by optimizing functional groups on our previously reported PI3Kα inhibitor. All the target compounds were synthesized and structural optimization on the structure of the lead compound based on cytotoxic activity. The results showed that some of the target compounds exhibited moderate to high cytotoxic activity against cell line U87MG and PC-3. The activities against mTOR kinase were investigated and the compound 12q showed excellent activity with an IC50 value of 54 nM in the same level of the positive control BEZ235 with IC50 value of 55 nM under the same test conditions. The western blot and cell cycle results demonstrate that compound 12q is a candidate as an mTORC1/mTORC2 dual-target inhibitor. The theoretical calculations were also performed to better understanding the binding modes of the compound 12q in the mTOR active site.

Design, Synthesis and Biological Evaluation of 4-Aryl-5,7-dihydro-6H-pyrrolo[2,3-d]pyrimidin-6-one Derivatives as a PI3Kα Inhibitor.

A novel series of 4-aryl-5,7-dihydro-6H-pyrrolo[2,3-d]pyrimidin-6-one derivatives were designed as a phosphoinositide 3-kinase α (PI3Kα) inhibitor by scaffold hopping. The target compounds, characterized by 1H-NMR, 13C-NMR and high resolution (HR)-MS, were synthesized from diethyl malonate and ethyl chloroacetate by nucleophilic substitution, ring-closure, chlorination and Suzuki reaction, etc. The biological activities were evaluated with cytotoxic activity in vitro on Uppsala 87 Malignant Glioma (U87MG) and prostate cancer-3 (PC-3) by Cell Counting Kit-8 (CCK-8). The results showed that compound 9c displayed the higher inhibition than the positive control PI-103, and high PI3Kα inhibitory activity with IC50 of 113 ± 9 nM in the same order of magnitude as BEZ235. In addition, the Log Kow values and molecular docking studies were performed to further investigate the drug-like properties of target compounds and interactions between 9c and PI3Kα.

Treatment options and clinical outcomes for carbapenem-resistant Enterobacteriaceae bloodstream infection in a Chinese university hospital.

BACKGROUND: Carbapenem resistant Enterobacteriaceae (CRE) has become a serious public health problem. Limited information is available about the treatment options that physicians used to fight CRE infections and related clinical outcomes in China. The aim of the present study was to explore the treatment options and clinical outcomes of patients with CRE bloodstream infection (BSI) in a Chinese teaching hospital. METHODS: A retrospective study was conducted during 2011 to 2015 in one Chinese teaching hospital. Demographic, microbiological and clinical characteristics of enrolled subjects were collected from medical records. Data were analyzed by Kaplan-Meier graphs, log-rank test, and Cox regression. RESULTS: A total of 98 inpatients with CRE BSI were enrolled in this study. For empirical therapy, 26 patients (26.5%) received at least one active drug within 48h after the onset of bacteremia. For definitive treatment, 59.2% (49/82) patients received at least one active drug and 40.2% (33/82) patients received therapy with no active drug. The overall 30-day mortality was 53.1% (52/98). Adverse outcome appeared to be more likely among patients with previous carbapenem exposure, neutropenia, severity of septic and time to initiation of BSIs. There was no significant difference in the mortality between the two groups of patients with combination therapy versus monotherapy (p=0.105). Severity of sepsis and neutropenia were identified as independent predictors of mortality. CONCLUSIONS: Our study demonstrated a high mortality associated with CRE BSI and a high percentage of inappropriate empirical treatment for CRE BSI patients in a Chinese teaching hospital. Particular attention should be given to the patients with CRE BSI.

Evaluation of automated systems for aminoglycosides and fluoroquinolones susceptibility testing for Carbapenem-resistant Enterobacteriaceae.

BACKGROUND: Automated systems (MicroScan WalkAway 96 Plus, Phoenix 100, and Vitek 2 Compact) are widely used in clinical laboratories nowadays. The aim of this study is to evaluate the performance of these three systems for susceptibility testing of aminoglycosides and fluoroquinolones against Carbapenem-resistant Enterobacteriaceae (CRE). METHODS: A total of 75 CRE isolates were used in this study. Quinolone resistance determinants (QRDs) (qnrA, qnrB, qnrC, qnrD, qnrS, aac(6')-Ib-cr, oqxAB and qepA) and aminoglycoside resistance determinants (ARDs) (aac(6')-Ib, armA, npmA, rmtA, rmtB, rmtC, rmtD and rmtE) of these CRE were screened by PCR. The MICs of aminoglycosides (gentamicin and amikacin) and fluoroquinolones (ciprofloxacin and levofloxacin) to CRE obtained with the automated systems were compared with the reference method (agar dilution method). RESULTS: Totally, 97.3% (73/75) of CRE harbored QRDs. The qnr gene was the most common QRD determinant identified in 68 (96.7%), followed by aac (6')-Ib-cr in 56 (74.7%), oqxAB in 23 (30.7%), and qepA in 2 (2.7%), respectively. 22.7% (17/75) of CRE harbored ARD determinants. rmtA, rmtB and npmA were identified among these isolates in 6 (8.0%), 6 (8.0%) and 5 (6.7%), respectively. A total of 900 results were obtained in this study. Overall, the total error rate was 9.89%. Twenty-eight very major errors (3.11%), 22 major errors (2.44%) and 39 minor errors (4.33%) were identified against agar dilution method. The very major errors were almost evenly distributed between results for fluoroquinolones (2.89%) and aminoglycosides (3.33%), while the major errors and minor errors were more commonly found in the results of fluoroquinolones (3.11% and 6.44%, respectively) than aminoglycosides (1.78% and 2.22%, respectively). CONCLUSIONS: Our study shows that testing difficulties in susceptibility testing do exist in automated systems. We suggest clinical laboratories using automated systems should consider using a second, independent antimicrobial susceptibility testing method to validate aminoglycosides and fluoroquinolones susceptibility.

Molecular characteristic of mcr-1 producing Escherichia coli in a Chinese university hospital.

BACKGROUND: Colistin has been considered as a last-line treatment option in severe infections caused by multidrug-resistant (MDR) gram-negative pathogens. However, the emergence of the mobile colistin resistance gene (mcr-1) has challenged this viewpoint. The aim of this study is to explore the prevalence of mcr-1 in Escherichia coli (E. coli) in a Chinese teaching hospital, and investigate their molecular characteristics. METHODS: A total of 700 E. coli isolates were used to screen mcr-1 by PCR and sequencing in a Chinese university hospital from August 2014 to August 2015. Susceptibility test of mcr-1-producing isolates was determined by Vitek -2 Compact system. 26 virulence factors (VFs), phylogenetic groups, Multi-locus sequence typing (MLST), and DNA Fingerprinting (ERIC-PCR) of strains were investigated by PCR. RESULTS: Four (0.6%) mcr-1 producing E. coli isolates were found in this study. The results of antibiotic susceptibility test showed that all four isolates were resistant to colistin, ciprofloxacin, levofloxacin, cefazolin, and trimethoprim/sulfamethoxazole, and were susceptible to amikacin, ertapenem and imipenem. In addition, all 4 isolates exhibited high-level resistance to aztreonam, cefotaxime and gentamicin. The numbers of VFs contained in mcr-1 positive isolates were no more than 4 in our study. MLST result demonstrated that these isolates were assigned to two sequence types: ST156 and ST167. The result of phylogenetic analysis showed that four mcr-1-positive isolates belong to two phylogenetic groups: A and B1 group. ERIC-PCR showed that four mcr-1 positive strains were categorized into three different genotypes. CONCLUSIONS: Our study demonstrated a low prevalence of mcr-1 in E. coli clinical isolates in a Chinese teaching hospital, and we have gained insights into the molecular characteristics of these mcr-1-positive strains. Increasing the surveillance of these infections, as well as taking effective infection control measures are urgently needed to take to control the transmission of mcr-1 gene.

High prevalence of metallo-ß-lactamase among carbapenem-resistant Klebsiella pneumoniae in a teaching hospital in China.

The aim of this study was to characterize the carbapenemases in carbapenem-resistant Klebsiella pneumoniae (CR-KP) from a Chinese teaching hospital. A total of 40 CR-KPs were screened for the presence of carbapenemases. Minimum inhibitory concentrations were determined by agar dilution. The modified Hodge test was used for the detection of carbapenemase production. Carbapenemase, extended-spectrum ß-lactamase, and AmpC genes were detected using polymerase chain reaction (PCR) and sequencing. A conjugation test was performed using a broth culture mating method, transferred plasmids were typed by PCR-based replicon typing, and clonal relatedness was investigated by enterobacterial repetitive intergenic consensus sequences PCR (ERIC-PCR) and multilocus sequence typing (MLST). The results revealed that modified Hodge test was positive for 28 CR-KPs, and CR-KPs exhibited high resistance rates against various antibiotics, except colistin (5.0%) and tigecycline (22.5%). ERIC and MLST profiles showed no clonal outbreak. PCR demonstrated a high prevalence rate (55.0%, 22/40) of metallo-ß-lactamases (MBLs) in CR-KPs. IMP-4, IMP-8, NDM-1, and KPC-2 were identified in 14 (35.0%), 7 (17.5%), 2 (5.0%), and 7 (17.5%) isolates, respectively. Notably, 2 CR-KPs coproduced 2 carbapenemases simultaneously (IMP-8/NDM-1 and IMP-4/KPC-2). In vitro transfer of carbapenem resistance was successful for 11 MBL-producing CR-KPs. The extended spectrum ß-lactamase genes were detected in 30 (75.0%) of these CR-KPs. To the best of our knowledge, this is the first report focusing on carbapenem resistance in K. pneumoniae due to metalloenzymes in China. Screening and surveillance of MBLs in Enterobacteriaceae is urgently needed in this region to control and prevent the spread of these resistance determinants.

Fecal carriage of carbapenem-resistant Enterobacteriaceae in a Chinese university hospital.

BACKGROUND: Carbapenem-resistant Enterobacteriaceae (CRE) is widespread in China. To date, no study available has specifically determined the prevalence and risk factors of inpatients with CRE intestinal colonization in this region. METHODS: Stool samples were screened for the presence of CRE in a Chinese university hospital. A case-control study was performed to identify risk factors associated with CRE fecal colonization. Case patients were those who had CRE colonization. Control subjects had no microbiologic evidence of CRE colonization. Clinical data were obtained from the medical record. RESULTS: The prevalence of CRE was 6.6% (20/303 patients), of which 8 had carbapenemase-producing isolates. KPC-2, IMP-4, and NDM-1 were detected from these isolates. Hospital readmissions (odds ratio [OR], 58.067; 95% confidence interval [95% CI]: 5.517-611.134; P = .001), sickbed changes (OR, 45.904; 95% CI: 8.484-248.376; P < .001), invasive procedures (OR, 8.322; 95% CI: 1.996-34.690; P = .004), and vancomycin (OR, 11.552; 95% CI: 1.155-115.574; P = .037) were independently associated with CRE colonization. CONCLUSION: This study demonstrated that asymptomatic intestinal carriage of CRE was relatively common in one region of China. Our study suggested that the implementation of effective infection control measures is urgently required to control the transmission of CRE in health care facilities in this country.