Objective To evaluate the diagnostic performance of plasma miR-499 in acute myocardial infarction (AMI) diagnosis.Methods Diagnostic accuracy test.The suspected AMI patients,who with chest pain for more than half an hour and been admitted in the Second People's Hospital of Wuxi and First People's Hospital of chuzhou during October 2010 and July 2011,were consecutively and prospectively enrolled in the present study.Sixty apparently healthy individuals were designed as healthy control.The plasma samples of the suspected AMI patients were collected within two hours after admission.The plasma miR-499 was determined by real time polymerase chain reaction (RT-PCR).The diagnostic performance of plasma miR-499 for AMI was estimated by receiver operating characteristic (ROC) curve analysis.The association between plasma miR-499 and AMI was analyzed by multivariable logistic model.The plasma miR-499 was determined and explained in blind fashion.Results Two hundred and nine suspected AMI patients,including 131 confirmed AMI patients (46 STEMI and 85 NSTEMI) and 78 AMI free patients were enrolled in the present study.The delta cycle threshold (ΔCT) was 1.01 ± 3.34 for AMI patients,-2.76 ± 2.90 for non-AMI patients and-3.79 ± 2.21 for healthy controls.The differences had statistical significance (x2 =96.77,P ＜ 0.01).The area under curve (AUC) of plasma miR-499 was 0.80 (95％ C I:0.74-0.86),lower than that of cardiac troponin Ⅰ (AUC =0.90,95％ CI:0.86-0.94) on admission (P ＜0.01).At the optimal cut-off of 0.18,the diagnostic sensitivity and specificity were 0.69 (95％ CI:0.61-0.77) and 0.77 (95％ CI:0.66-0.86),respectively.The coefficient of correlation between plasma miR-499 and cTnI was 0.72 (P ＜0.01).The odds ratio (OR) of plasma miR-499 ＞0.18 for AMI was 2.59 (95％ CI:1.10-6.07),after adjusted cTnI.Conclusions Plasma miR-499 is a useful biomarker for AMI diagnosis.It can provide additional diagnostic information beyond cTnI.Combination utility of plasma miR-499 and cTnI may improve the diagnostic accuracy for AMI.

ObjectiveTo evaluate the feasibility of constructing tissue engineering cardiac patch with photooxidationfixed acellular bovine pericardium.MethodsFresh bovine pericardia were treated by dye-mediated photooxidation after decellularization.Some of them were seeded with bone marrow stromal cells(MSCs) isolated from male SD rats to construct cardiac patches.Myocardial infarction(MI) model was made in female SD rats by left anterior descending coronary ligation(LAD).One week later, the confirmed MI rats were divided into three groups randomly, group MI (n = 15)without any treatment; group P (n = 18) with photooxidated pericardia implantation ; group P + C (n = 18) with seeded pericardia implantation.A sham group (n = 10) was also performed with opening and closing chest twice only.The heart were explanted at 2 or 4 weeks after implantation, and examined histologically and immunohistochemically.The heart function was evaluated by echocardiography at 4 weeks before excising the rats.ResultsThere were no cells or cell debris remained in bovine pericardium tissue.The fiber structure became condensed after photooxidation.The seeded cells formed a continuous layer on the surface of the tissue.The pericardial degradation level and newly formed microvessel density were larger in group P + C than in group P after 2 [ (13.7 ±5.2)个/mm2 vs (7.1 ±3.1)个/mm2, P＜0.05]and4 [(22.6 ±4.9)个/mrn2 vs (14.1 ±5.3)个/mm2, P＜0.05]weeks.Four weeks after transplantation, cardiac echocardiography showed left ventricular ejection fraction(LVEF) was lower in group MI (44.8 ± 4.4) ％ and group P (48.4 ± 5.0) ％ compared with group P + C (49.3 ± 4.8) ％, left ventricular fractional shorterning(LVFS) was lower in group MI (18.0 ± 2.2) ％ and group P (19.8 ± 2.5) ％ compared with group P + C (20.4 ±2.5) ％, the difference between P + C and MI was significant.ConclusionTransplantation of the tissue engineered bovine pericardial patches with dye-mediated photooxidation can improve heart function in MI rats.This kind of patches demonstrates a promising prospect in the future.

BACKGROUND:During the outbreak of severe acute respiratory syndrome(SARS),certain people had been isolated by various reasons and appeared a series of psychological, physical and behavioral reactions. OBJECTIVE:To understand the different defensive features in people with different level of mental health in the isolated population, and explore the relationship between defensive style and mental health. DESIGN:An in investigative study taking the isolated population during outbreak of SARS as the subjects. SETTING:A psychiatric department of a military hospital. PARTICIPANTS:Totally 187 people of different sex,age and education, who were isolated during April and May 2003 due to SARS outbreak in a city of northwest China,were selected as the subjects. INTERVENTIONS:The 187 subjects,who were isolated due to SARS outbreak,were evaluated by using the symptom checklist(SCL 90) and defense style questionnaire(DSQ). RESULTS:About 36.4% people of this population had distinct mental or physical health problems that were characterized by anxiety,horror,depression,hostility and compulsion.There was difference in defensive styles between the high symptom group and low symptom group,among which the score of DSQ factors in the immature type of high and low symptom groups were 5.72± 1.56 and 4.35± 0.96 respectively while the scores in the intermediate type defense were 4.98± 1.44 and 3.72± 0.89 respectively(P< 0.01).Mental health problem was positively correlated with the application of immature defensive style,but had negative correlation with application of mature defensive means. CONCLUSION:There is difference in the defensive styles among the isolated people with different mental health status,and their defensive strategies are closely related to the mental health.

Objective To explore the characteristic manifestations of breast malignant isodense masses using mammography. Methods 121 breast isodense masses with pathological confirmations were collected.Compared indicators with pathological findings were the tumor size,shape,edge,structure changes around the mass,axillary lymph nodes,skin changes,nipple changes,suspi-cious malignant calcification.χ2 test and Logistic regression analysis were used to evaluate the diagnostic values of each indicator.Re-sults In the 121 isodense masses,73 were benign,48 were malignant tumors.According to χ2 test analysis,factors impacted the nature of mass including:tumor morphology (χ2 =14.376,P =0.002),tumor edge (χ2 = 21.555,P =0.000),structure twisted surrounding the mass (χ2 =26.939,P =0.000),axillary lymph nodes (χ2 =1 6.285,P =0.000),skin thickening (χ2 =4.698,P =0.030).According to multivariate binary Logistic regression analysis,risk factors of the Logistic regression model included:edge infiltrating of mass (P =0.014),structure twisted surrounding the mass (P =0.003),axillary lymph nodes (P =0.026).Conclusion The characteristic manifestations of breast malignant isodense masses include edge infiltrating,structure twisted surrounding the mass,axillary lymph nodes.These manifestations are of great significance in differential diagnosis of breast isodense masses.

Objective To explore the brain electrophysiological mechanism of object rotation in first-episode schizophrenia.Methods 30 patients with schizophrenia and 28 normal healthy people,who were from the Center for Mental Disease Control and Prevention,Third Hospital of PLA,took part in the mental rotation tasks,then the incubation period and amplitude of P500,and the wrong number and reaction time were measured.Results Compared with control group ( normal:(494.16 ± 34.68 ) ms,( 9.56 ± 2.54) μV; mirror:(496.51 ± 33.10 ) ms,(6.38 ± 2.41 ) μV),schizophrenia' incubation periods were significantly delayed ( normal:( 571.30 ± 51.21 ) ms;mirror:(573.41 ±39.27) ms) and volatility were significantly lower ( normal:(4.26 ± 1.01 ) μV; mirror:(3.61± 1.21 )μV) in normal and mirror rotation (P＜0.05 ).The mirror-normal differences were not significant on the incubation periods of two groups (P ＞ 0.05 ) ; the mirror-normal image differences were not significant on the patient group' volatility (P ＞ 0.05 ) ; the normal volatility was significantly higher than mirror in the control group (P ＜ 0.05).Conclusion Schizophrenia'mental rotation ability is impaired,and mirror-normal differences on mental rotation are disappeared.It can be used as an early-stage clinical auxiliary diagnosis index.

Dehydration is one of the key steps in the biosynthesis of mycolic acids and is vital to the growth of Mycobacterium tuberculosis (Mtb). Consequently, stalling dehydration cures tuberculosis (TB). Clinically used anti-TB drugs like thiacetazone (TAC) and isoxyl (ISO) as well as flavonoids inhibit the enzyme activity of the β-hydroxyacyl-ACP dehydratase HadAB complex. How this inhibition is exerted, has remained an enigma for years. Here, we describe the first crystal structures of the MtbHadAB complex bound with flavonoid inhibitor butein, 2',4,4'-trihydroxychalcone or fisetin. Despite sharing no sequence identity from Blast, HadA and HadB adopt a very similar hotdog fold. HadA forms a tight dimer with HadB in which the proteins are sitting side-by-side, but are oriented anti-parallel. While HadB contributes the catalytically critical His-Asp dyad, HadA binds the fatty acid substrate in a long channel. The atypical double hotdog fold with a single active site formed by MtbHadAB gives rise to a long, narrow cavity that vertically traverses the fatty acid binding channel. At the base of this cavity lies Cys61, which upon mutation to Ser confers drug-resistance in TB patients. We show that inhibitors bind in this cavity and protrude into the substrate binding channel. Thus, inhibitors of MtbHadAB exert their effect by occluding substrate from the active site. The unveiling of this mechanism of inhibition paves the way for accelerating development of next generation of anti-TB drugs.
Amino Acid Sequence ; Bacterial Proteins ; chemistry ; metabolism ; Catalytic Domain ; Enzyme Inhibitors ; chemistry ; pharmacology ; Flavonoids ; chemistry ; pharmacology ; Hydro-Lyases ; antagonists & inhibitors ; chemistry ; Molecular Sequence Data ; Mycobacterium tuberculosis ; drug effects ; enzymology ; Protein Binding ; drug effects ; Protein Multimerization ; drug effects ; Protein Structure, Secondary ; Sequence Alignment

During severe acute respiratory syndrome coronavirus (SARS-CoV) infection, the activity of the replication/transcription complexes (RTC) quickly peaks at 6 hours post infection (h.p.i) and then diminishes significantly in the late post-infection stages. This "down-up-down" regulation of RNA synthesis distinguishes different viral stages: primary translation, genome replication, and finally viron assembly. Regarding the nsp8 as the primase in RNA synthesis, we confirmed that the proteolysis product of the primase (nsp8) contains the globular domain (nsp8C), and indentified the resectioning site that is notably conserved in all the three groups of coronavirus. We subsequently crystallized the complex of SARS-CoV nsp8C and nsp7, and the 3-D structure of this domain revealed its capability to interfuse into the hexadecamer super-complex. This specific proteolysis may indicate one possible mechanism by which coronaviruses to switch from viral infection to genome replication and viral assembly stages.
Amino Acid Sequence ; Crystallography, X-Ray ; DNA Primase ; chemistry ; genetics ; physiology ; Humans ; Isoenzymes ; chemistry ; genetics ; physiology ; Molecular Sequence Data ; Protein Structure, Secondary ; RNA, Viral ; biosynthesis ; SARS Virus ; chemistry ; genetics ; physiology ; Sequence Alignment ; Severe Acute Respiratory Syndrome ; virology ; Virus Replication

Disulfide bond-forming (Dsb) protein is a bacterial periplasmic protein that is essential for the correct folding and disulfide bond formation of secreted or cell wallassociated proteins. DsbA introduces disulfide bonds into folding proteins, and is re-oxidized through interaction with its redox partner DsbB. Mycobacterium tuberculosis, a Gram-positive bacterium, expresses a DsbA-like protein ( Rv2969c), an extracellular protein that has its Nterminus anchored in the cell membrane. Since Rv2969c is an essential gene, crucial for disulfide bond formation, research of DsbA may provide a target of a new class of anti-bacterial drugs for treatment of M.tuberculosis infection. In the present work, the crystal structures of the extracellular region of Rv2969c (Mtb DsbA) were determined in both its reduced and oxidized states. The overall structure of Mtb DsbA can be divided into two domains: a classical thioredoxin-like domain with a typical CXXC active site, and an α-helical domain. It largely resembles its Escherichia coli homologue EcDsbA, however, it possesses a truncated binding groove; in addition, its active site is surrounded by an acidic, rather than hydrophobic surface. In our oxidoreductase activity assay, Mtb DsbA exhibited a different substrate specificity when compared to EcDsbA. Moreover, structural analysis revealed a second disulfide bond in Mtb DsbA, which is rare in the previously reported DsbA structures, and is assumed to contribute to the overall stability of Mtb DsbA. To investigate the disulphide formation pathway in M.tuberculosis, we modeled Mtb Vitamin K epoxide reductase (Mtb VKOR), a binding partner of Mtb DsbA, to Mtb DsbA.
Amino Acid Sequence ; Bacterial Proteins ; chemistry ; metabolism ; Catalytic Domain ; Crystallography, X-Ray ; Disulfides ; chemistry ; Escherichia coli ; metabolism ; Escherichia coli Proteins ; chemistry ; metabolism ; Molecular Docking Simulation ; Molecular Sequence Data ; Mycobacterium tuberculosis ; metabolism ; Oxidation-Reduction ; Protein Disulfide-Isomerases ; chemistry ; metabolism ; Protein Folding ; Protein Structure, Tertiary ; Sequence Alignment ; Static Electricity

The guanine-nucleotide exchange factor (GEF) RalGPS1a activates small GTPase Ral proteins such as RalA and RalB by stimulating the exchange of Ral bound GDP to GTP, thus regulating various downstream cellular processes. RalGPS1a is composed of an Nterminal Cdc25-like catalytic domain, followed by a PXXP motif and a C-terminal pleckstrin homology (PH) domain. The Cdc25 domain of RalGPS1a, which shares about 30% sequence identity with other Cdc25-domain proteins, is thought to be directly engaged in binding and activating the substrate Ral protein. Here we report the crystal structure of the Cdc25 domain of RalGPS1a. The bowl shaped structure is homologous to the Cdc25 domains of SOS and RasGRF1. The most remarkable difference between these three Cdc25 domains lies in their active sites, referred to as the helical hairpin region. Consistent with previous enzymological studies, the helical hairpin of RalGPS1a adopts a conformation favorable for substrate binding. A modeled RalGPS1a-RalA complex structure reveals an extensive binding surface similar to that of the SOS-Ras complex. However, analysis of the electrostatic surface potential suggests an interaction mode between the RalGPS1a active site helical hairpin and the switch 1 region of substrate RalA distinct from that of the SOS-Ras complex.
Amino Acid Sequence ; Binding Sites ; Catalytic Domain ; Cloning, Molecular ; Crystallography, X-Ray ; Escherichia coli ; Guanosine Diphosphate ; metabolism ; Guanosine Triphosphate ; metabolism ; Humans ; Models, Molecular ; Molecular Conformation ; Molecular Sequence Data ; Plasmids ; metabolism ; Protein Binding ; Protein Structure, Tertiary ; genetics ; Recombinant Proteins ; chemistry ; genetics ; metabolism ; ral GTP-Binding Proteins ; chemistry ; genetics ; metabolism ; ral Guanine Nucleotide Exchange Factor ; chemistry ; genetics ; metabolism

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