Objective To explore strategies and measures to improve hand hygiene (HH)compliance and correctness of health care workers (HCWs)in a large hospital.Methods The WHO multimodal hand hygiene improvement strategy was adopted by healthcare-associated infection(HAI)management department of a hospital,measures consisted of five key com-ponents,including:system change,education and training,evaluation and feedback,reminders in the workplace,and insti-tutional safety climate.HH compliance and correctness of HCWs were observed by infection control practitioners,HH compliance and correctness in January-June of 2012 (pre-intervention)and January-June of 2014 (post-intervention)were compared,effectiveness of intervention strategies were evaluated.Results HH compliance rate and correctness rate of post-intervention were both higher than pre-intervention (75.92% [8 369/11 023]vs 53.67% [5 127/9 553],P <0.001;94.11%[7 782/8 269]vs 83.88%[3 642/4 342],P <0.001).Of different occupations,HH compliance rates of doctors and nurses were significantly different before and after intervention (both P <0.001),while workers and staff of other oc-cupations before and after intervention were not significantly different(both P >0.05).Except ‘after body fluid exposure’, HH compliance rates of the other four indications for HH before and after intervention were significantly different(all P <0.001).Conclusion HH compliance and correctness of HCWs can be improved after adopting WHO multimodal hand hy-giene improvement strategy.

Objective To study the clinical efficacy and operative skill of multi-endoscopic technique in treatment of post-traumatic urethrostenosis in male patients.Methods A retrospective analysis was done on clinical data of 47 male patients with post-traumatic urethrostenosis treated with direct visional incision urethrotomy combined with transurethral resection of scar tissue and ureteroscopic incision.There were 29 patients with anterior urethral strictures and 18 with posterior urethral strictures.Results Of all,43 patients underwent successful endoscopic surgeries at the first time but one underwent secondary surgery because of unsuccessful endoscopic incision.These patients achieved satisfactory results without urinary incontinence,fistula or reoperation.The left three patients underwent open surgeries because of unsuccessful endoscopic incision,in which one patient could micturate at maximal flow rate of 9-12 ml/s,without therapeutic urethral dilation,one could micturate under regular therapeutic dilation and the other one could not micturate.Conclusions With the advantages of safety,high success rate and good long-term efficiency,multi-endoscopic technique can be used as an initial treatment for male patients with post-traumatic urethrostenosis and is worthy to be popularized.

Objective To assess the clinical value of intravesical prostatic protrusion (IPP) measured by transabdominal uhrasonography in judgement of benign prostatic obstruction (BPO). Methods According to pressure-flow study, 109 patients with benign prostatic hyperplasia were di-vided into 3 groups (non-obstruction, equivocal obstruction and obstruction). IPP was measured by transabdominal ultrasonography in all patients. The difference of IPP between different groups was studied and the correlation between IPP and the parameters reflecting BPO was analyzed. Results IPP value of the non-obstruction group, equivocal obstruction group and obstruction group was 2.7 ±1.2 mm, 2.9±1.4 mm and 15.4±6.5 mm. There was significant difference in IPP between the non-obstruction group, equivocal obstruction group and obstruction group (P<0.01). IPP was correlated with the parameters including Pdet. Qbeg, Pdet. Qmax, Pdet. Qend, Qmax and A-G number, Spearman's ratio was 0. 628, 0. 714, 0. 591, -0. 450 and 0. 729(P<0.01), respectively. The sensi-tivity and specificity of judging BPO were 89.9% and 97.5% if the cut-off was IPP≥10 mm. Con-clasion IPP measured by transabdominal ultrasonography could be a convenient and accurate method in diagnosis of BPO.

Fusarium graminearum (sexual stage: Gibberella zeae) is the causative agent of Fusarium Head Blight (FHB), which is one of the most destructive plant disease of cereals, accounting for high grain yield losses, especially for wheat and maize. Like other fungal pathogens, several extracellular enzymes secreted by G. zeae are known to be involved in host infection. Among these secreted lipases, G. zeae lipase (GZEL), which is encoded by the FGL1 gene, was demonstrated to be crucial to G. zeae pathogenicity. However, the precise mechanism of GZEL remains unclear due to a lack of detailed structural information. In this study, we report the crystal structure of GZEL at the atomic level. The structure of GZEL displays distinct structural differences compared to reported homologues and indicates a unique "double lock" enzymatic mechanism. To gain insight into substrate/inhibitor recognition, we proposed a model of GZEL in complex with substrate and the lipase inhibitor ebelactone B (based on the reported structures of GZEL homologues), which defines possible substrate binding sites within the catalytic cleft and suggests an "anti sn-l" binding mode. These results pave the way to elucidating the mechanism of GZEL and thus provide clues for the design of anti-FHB inhibitors.
Amino Acid Sequence ; Catalytic Domain ; Crystallography, X-Ray ; Gibberella ; enzymology ; Lactones ; chemistry ; Lipase ; chemistry ; metabolism ; Models, Molecular ; Molecular Sequence Data ; Oleic Acid ; chemistry ; Protein Binding ; Protein Structure, Secondary ; Protein Structure, Tertiary ; Sequence Alignment ; Sequence Homology, Amino Acid ; Substrate Specificity ; Surface Properties

Coronaviruses are the causative agent of respiratory and enteric diseases in animals and humans. One example is SARS, which caused a worldwide health threat in 2003. In coronaviruses, the structural protein N (nucleocapsid protein) associates with the viral RNA to form the filamentous nucleocapsid and plays a crucial role in genome replication and transcription. The structure of N-terminal domain of MHV N protein also implicated its specific affinity with transcriptional regulatory sequence (TRS) RNA. Here we report the crystal structures of the two proteolytically resistant N- (NTD) and C-terminal (CTD) domains of the N protein from murine hepatitis virus (MHV). The structure of NTD in two different crystal forms was solved to 1.5 Å. The higher resolution provides more detailed structural information than previous reports, showing that the NTD structure from MHV shares a similar overall and topology structure with that of SARS-CoV and IBV, but varies in its potential surface, which indicates a possible difference in RNA-binding module. The structure of CTD was solved to 2.0-Å resolution and revealed a tightly intertwined dimer. This is consistent with analytical ultracentrifugation experiments, suggesting a dimeric assembly of the N protein. The similarity between the structures of these two domains from SARS-CoV, IBV and MHV corroborates a conserved mechanism of nucleocapsid formation for coronaviruses.
Amino Acid Sequence ; Binding Sites ; Crystallography, X-Ray ; Molecular Sequence Data ; Murine hepatitis virus ; chemistry ; metabolism ; Nucleocapsid Proteins ; chemistry ; metabolism ; Phosphoproteins ; chemistry ; metabolism ; Protein Binding ; Protein Folding ; Protein Multimerization ; Protein Structure, Secondary ; Protein Structure, Tertiary ; RNA ; metabolism ; Sequence Alignment

Binding Sites ; Cysteine Endopeptidases ; metabolism ; Humans ; Isoxazoles ; chemistry ; pharmacology ; Protease Inhibitors ; chemistry ; metabolism ; pharmacology ; Protein Structure, Tertiary ; Pyrrolidinones ; chemistry ; pharmacology ; Rhinovirus ; drug effects ; SARS Virus ; drug effects ; enzymology ; Severe Acute Respiratory Syndrome ; virology ; Viral Proteins ; antagonists & inhibitors ; metabolism

Gram-negative Enterobacteriaceae with resistance to carbapenem conferred by New Delhi metallo-β-lactamase 1 (NDM-1) are a type of newly discovered antibioticresistant bacteria. The rapid pandemic spread of NDM-1 bacteria worldwide (spreading to India, Pakistan, Europe, America, and Chinese Taiwan) in less than 2 months characterizes these microbes as a potentially major global health problem. The drug resistance of NDM-1 bacteria is largely due to plasmids containing the blaNDM-1 gene shuttling through bacterial populations. The NDM-1 enzyme encoded by the blaNDM-1 gene hydrolyzes β-lactam antibiotics, allowing the bacteria to escape the action of antibiotics. Although the biological functions and structural features of NDM-1 have been proposed according to results from functional and structural investigation of its homologues, the precise molecular characteristics and mechanism of action of NDM-1 have not been clarified. Here, we report the three-dimensional structure of NDM-1 with two catalytic zinc ions in its active site. Biological and mass spectroscopy results revealed that D-captopril can effectively inhibit the enzymatic activity of NDM-1 by binding to its active site with high binding affinity. The unique features concerning the primary sequence and structural conformation of the active site distinguish NDM-1 from other reported metallo-β-lactamases (MBLs) and implicate its role in wide spectrum drug resistance. We also discuss the molecular mechanism of NDM-1 action and its essential role in the pandemic of drug-resistant NDM-1 bacteria. Our results will provide helpful information for future drug discovery targeting drug resistance caused by NDM-1 and related metallo-β-lactamases.
Amino Acid Sequence ; Anti-Bacterial Agents ; metabolism ; Binding Sites ; Captopril ; chemistry ; pharmacology ; Catalytic Domain ; Crystallography, X-Ray ; Drug Resistance, Bacterial ; Enterobacteriaceae ; enzymology ; Molecular Sequence Data ; Sequence Alignment ; Sequence Homology, Amino Acid ; beta-Lactamases ; chemistry ; metabolism

Ebola virus (EBOV) is a key member of Filoviridae family and causes severe human infectious diseases with high morbidity and mortality. As a typical negative-sense single-stranded RNA (-ssRNA) viruses, EBOV possess a nucleocapsid protein (NP) to facilitate genomic RNA encapsidation to form viral ribonucleoprotein complex (RNP) together with genome RNA and polymerase, which plays the most essential role in virus proliferation cycle. However, the mechanism of EBOV RNP formation remains unclear. In this work, we solved the high resolution structure of core domain of EBOV NP. The polypeptide of EBOV NP core domain (NP(core)) possesses an N-lobe and C-lobe to clamp a RNA binding groove, presenting similarities with the structures of the other reported viral NPs encoded by the members from Mononegavirales order. Most strikingly, a hydrophobic pocket at the surface of the C-lobe is occupied by an α-helix of EBOV NP(core) itself, which is highly conserved among filoviridae family. Combined with other biochemical and biophysical evidences, our results provides great potential for understanding the mechanism underlying EBOV RNP formation via the mobility of EBOV NP element and enables the development of antiviral therapies targeting EBOV RNP formation.
Crystallography, X-Ray ; Ebolavirus ; physiology ; Humans ; Nucleoproteins ; chemistry ; genetics ; metabolism ; Protein Structure, Tertiary ; Structure-Activity Relationship ; Virus Assembly ; physiology

Unlike the well-established picture for the entry of enveloped viruses, the mechanism of cellular entry of non-enveloped eukaryotic viruses remains largely mysterious. Picornaviruses are representative models for such viruses, and initiate this entry process by their functional receptors. Here we present the structural and functional studies of SCARB2, a functional receptor of the important human enterovirus 71 (EV71). SCARB2 is responsible for attachment as well as uncoating of EV71. Differences in the structures of SCARB2 under neutral and acidic conditions reveal that SCARB2 undergoes a pivotal pH-dependent conformational change which opens a lipid-transfer tunnel to mediate the expulsion of a hydrophobic pocket factor from the virion, a pre-requisite for uncoating. We have also identified the key residues essential for attachment to SCARB2, identifying the canyon region of EV71 as mediating the receptor interaction. Together these results provide a clear understanding of cellular attachment and initiation of uncoating for enteroviruses.
Acids ; chemistry ; Amino Acid Sequence ; Animals ; Capsid Proteins ; chemistry ; genetics ; metabolism ; Enterovirus A, Human ; genetics ; metabolism ; physiology ; HEK293 Cells ; Host-Pathogen Interactions ; Humans ; Hydrogen-Ion Concentration ; Lysosome-Associated Membrane Glycoproteins ; chemistry ; genetics ; metabolism ; Molecular Docking Simulation ; Molecular Sequence Data ; Protein Binding ; Protein Conformation ; Protein Interaction Mapping ; Protein Structure, Tertiary ; RNA, Viral ; genetics ; metabolism ; Receptors, Scavenger ; chemistry ; genetics ; metabolism ; Sequence Homology, Amino Acid ; Sf9 Cells ; Static Electricity ; Virion ; genetics ; metabolism ; Virus Attachment

Enterovirus 71 (EV71), one of the major causative agents for hand-foot-and-mouth disease (HFMD), has caused more than 100 deaths among Chinese children since March 2008. The EV71 genome encodes an RNAdependent RNA polymerase (RdRp), denoted 3D(pol), which is central for viral genome replication and is a key target for the discovery of specific antiviral therapeutics. Here we report the crystal structures of EV71 RdRp (3D(pol)) and in complex with substrate guanosine-5'-triphosphate and analog 5-bromouridine-5'-triphosphate best to 2.4 Å resolution. The structure of EV71 RdRp (3D(pol)) has a wider open thumb domain compared with the most closely related crystal structure of poliovirus RdRp. And the EV71 RdRp (3D(pol)) complex with GTP or Br-UTP bounded shows two distinct movements of the polymerase by substrate or analogue binding. The model of the complex with the template:primer derived by superimposition with foot-and-mouth disease virus (FMDV) 3D/RNA complex reveals the likely recognition and binding of template:primer RNA by the polymerase. These results together provide a molecular basis for EV71 RNA replication and reveal a potential target for anti-EV71 drug discovery.
Amino Acid Sequence ; Child ; China ; epidemiology ; Crystallography, X-Ray ; Drug Discovery ; Enterovirus A, Human ; chemistry ; enzymology ; Hand, Foot and Mouth Disease ; drug therapy ; epidemiology ; virology ; Humans ; Models, Molecular ; Molecular Sequence Data ; Molecular Targeted Therapy ; Protein Conformation ; Protein Folding ; RNA Replicase ; chemistry ; genetics ; metabolism ; Sequence Alignment ; Substrate Specificity