[Effect of neuregulin-1 on cardiac glucose metabolism in rats with experimental myocardial infarction].

Objective: To investigate the effect of neuregulin-1(NRG-1) on cardiac glucose metabolism in Sprague Dawley (SD) rats with experimental myocardial infarction (MI). Methods: Adult male SD rats were randomly divided into three groups: the sham-operated group, MI group, and MI+NRG1 group. The rat MI model was established via ligation of the left anterior descending coronary artery. Two weeks after operation, echocardiography was performed, MI rats with left ventricular ejection fraction (LVEF) between 0.3-0.5 were selected and randomly assigned to MI group and MI+NRG-1 group. Rats in MI+NRG-1 group were treated with recombinant human NRG-1ß (100 µg/kg) via tail vein at 2 weeks after operation (twice per week for 6 weeks); while rats in sham-operated group and MI group received equal volume of physiological saline. By the end of administration, echocardiography and small animal positron emission tomography (PET) were performed to detect cardiac function and myocardial glucose uptake. Myocardial morphology and collagen volume fraction, cardiomyocyte apoptosis and reactive oxygen species (ROS) production were evaluated by histopathologic analysis. Myocardial pyruvate dehydrogenase (PDH) and citrate synthase (CS) activity, as well as ATP production were detected by commercial kits. The mRNA and protein expression levels of NRG-1, p-ErbB4, and key factors involved in glucose metabolism (including Glut-4, HK2, PDK4, PDH, CS) were detected by quantitative real-time PCR (qRT-PCR) and Western blot assay, respectively. Results: With the MI model successfully established, the left ventricular ejection fraction(LVEF) and left ventricular shortening fraction(LVFS) were significantly lower in MI group and MI+NRG-1 group than that in sham group (both P<0.01), while there was no significant difference between MI group and MI+NRG-1 group(all P>0.05). After 6 weeks of NRG-1ß intervention, the LVEF and LVFS were significantly higher in MI+NRG-1 group than in MI group (both P<0.01). By the end of experiment, PET imaging showed that the mean standardized uptake value (SUVmean) were lower in MI+NRG-1 group than in the sham group (4.06±0.28 vs. 5.18±0.37, P<0.01), while significantly higher than that in MI group (4.06±0.28 vs.2.86±0.49, P<0.01). Histopathological analysis showed that compared with MI group, rats in MI+NRG-1 group exhibited significantly decreased left ventricle collagen volume fraction ((7.83±1.24) % vs. (18.31±3.58) %, P<0.01), cardiomyocyte apoptosis((37.98±4.26)% vs. (67.04±5.38)%, P<0.01), and DHE fluorescence intensity(0.057 28±0.007 06 vs. 0.076 94±0.008 46, P<0.01), indicating that NRG-1ß could reduce ROS production. PDH activity, CS activity, and ATP production were significantly higher in MI+NRG-1 group than in MI group (all P<0.05). qRT-PCR demonstrated an upregulated Glut-4, HK2 and CS, but downregulated PDK4 mRNA expression in MI+NRG-1 group compared with MI group (all P<0.01). Western blot assay showed significantly higher protein expression of NRG-1, p-ErbB4, Glut-4, HK2, PDH, CS in MI+NRG-1 group than in MI group (all P<0.01). Conclusion: NRG-1 could improve glucose uptake and utilization in myocardium by activating phosphorylation of myocardial ErbB4 receptor in MI rats, thus providing a therapeutic option for improving energy metabolism after MI.

Dementia predicted one-year mortality for patients with first hip fracture: a population-based study.

Aims: We aimed to determine the effect of dementia and Parkinson's disease on one, three and 12-month mortality following surgery for fracture of the hip in elderly patients from an Asian population. Patients and Methods: Using a random sample of patients taken from the Taiwan National Health Insurance Research Database, this retrospective cohort study analyzed the data on 6626 elderly patients who sustained a fracture of the hip between 1997 and 2012 who had ICD-9 codes within the general range of hip fracture (820.xx). We used Cox regression to estimate the risk of death associated with dementia, Parkinson's disease or both, adjusting for demographic, clinical, treatment, and provider factors. Results: Among 6626 hip fracture patients, 10.20% had dementia alone, 5.60% had Parkinson's disease alone, and 2.67% had both. Corresponding one-year mortality rates were 15.53%, 11.59%, and 15.82%, compared with 9.22% for those without neurological illness. Adjusted hazard ratio for one-year mortality was 1.45 (95% confidence intervals (CI) 1.17 to 1.79) for those with dementia, and 1.57 (95% CI 1.07 to 2.30) with both dementia and Parkinson's disease versus patients with neither. There was no significant association with death for Parkinson's disease alone. Age, male gender and comorbidities were also associated with a higher risk of mortality. Conclusion: Dementia, with or without Parkinson's disease, is an independent predictor of mortality following surgery for fractures of the hip. Age, male gender and comorbidities also increase the risk of death. Parkinson's disease alone has no significant effect. Cite this article: Bone Joint J 2018;100-B:1220-6.

Atrial fibrillation prevalence and atrial vulnerability analysis in paroxysmal supraventricular tachycardia patients after radiofrequency ablation.

OBJECTIVE: Paroxysmal supraventricular tachycardia (PSVT) patient has higher prevalence ratio of atrial fibrillation (AF), while AF recurrence remains a high ratio after radiofrequency ablation (RFCA). This study explored AF prevalence and atrial vulnerability in PSVT patients after RFCA, using ROC curve to analyze atrial effective refractory period dispersion (dERP) on postoperative AF in the prediction of PSVT. PATIENTS AND METHODS: The PSVT patients were enrolled and divided into three groups and received RFCA treatment between January 2010 and December 2014. Group A, AF was induced by programmed electrical stimulation (PES) under isopropyl epinephrine (ISO) perfusion; group B, AF was induced by PES or PES under ISO perfusion; group C, no AF was generated by PES or PES under ISO perfusion. Ultrasonic cardiogram was applied to detect intra, inter, atrial electromechanical delay and dERP. ROC curve was adopted to analyze the prediction effect of dERP on postoperative AF recurrence. RESULTS: dERP, group B > A > C; inter and intra: group B > A (p<0.05). Logistic regression analysis showed that age, gender, sympathetic nerve stimulation, and left/right atrial diameter were independent risk factors of postoperative AF in PSVT patients (p<0.05). Of note, dERP cut-off value at 75.5 msec can effectively predict AF recurrence in PSVT patients after RFCA (p<0.05). CONCLUSIONS: Atrial vulnerability index dERP could be applied to evaluate AF recurrence in PSVT patients after RFCA.

Rosuvastatin alleviates the development of monocrotaline-induced pulmonary hypertension in rats.

Statins can increase endothelial function through enhancement of the expression and activity of endothelial nitric oxide synthase (eNOS). The aim of this study is to evaluate the effect of rosuvastatin on the number of circulating endothelial progenitor cells (EPCs) and endothelial expression of eNOS in monocrotaline-induced pulmonary hypertensive rats. Sixty Sprague-Dawley (SD) rats were divided into three groups of 20: control (group A), pulmonary hypertension (PAH) + rosuvastatin group (group B), and PAH (group C). Monocrotaline (MCT; 60 mg/kg) was injected (intraperitoneally) to induce PAH. Rats in group B received rosuvastatin [10 mg/(kg. day)] for 2 weeks. Peripheral blood (5 mL) was aspirated from the femoral artery of each rat before and after 2 weeks of treatment. Mononuclear cells were isolated and subcultured to obtain EPCs. Small and moderately sized pulmonary arteries were collected 2 weeks later for histological analyses. eNOS gene expression in endothelial cells of pulmonary arteries were then determined at mRNA and protein levels. eNOS expression at mRNA and protein levels and the number of circulating EPCs were reduced significantly in groups B and C compared with group A (P less than 0.05), and a significant difference between group B and group C (P less than 0.05) was observed. Vascular remodeling in small and moderately sized pulmonary arteries was attenuated markedly in group B compared with group C. These results suggest that rosuvastatin can ameliorate the remodeling of pulmonary arteries in MCT-induced PAH rats by increasing the number of circulating EPCs and eNOS upregulation.

Person perception precedes theory of mind: an event related potential analysis.

Prior to developing an understanding of another person's mental state, an ability termed "theory of mind" (ToM), a perception of that person's appearance and actions is required. However the relationship between this "person perception" and ToM is unclear. To investigate the time course of ToM and person perception, event-related potentials (ERP) were recorded while 17 normal adults received three kinds of visual stimuli: cartoons involving people (person perception cartoons), cartoons involving people and also requiring ToM for comprehension (ToM cartoons), and scene cartoons. We hypothesized that the respective patterns of brain activation would be different under these three stimuli, at different stages in time. Our findings supported this proposal: the peak amplitudes of P200 for scene cartoons were significantly lower than for person perception or ToM cartoons, while there were no significant differences between the latter two for P200. During the 1000-1300 ms epoch, the mean amplitudes of the late positive components (LPC) for person perception were more positive than for scene representation, while the mean amplitudes of the LPC for ToM were more positive than for person perception. The present study provides preliminary evidence of the neural dynamic that underlies the dissociation between person perception and ToM.

How to map ses, a mutant of Arabidopsis thaliana affecting pollen development.

In Arabidopsis, map-based cloning has been developed to an effective method in mutant genetic analysis because high-density markers are available, candidate genes or genomic sequences can be amplified by PCR and transgenic techniques are simplified. Mutant ses named from shortened early-stage siliques was used as an example to show how to map a mutant in this day. By the process of bulked segregants analysis, linkage testing, large-scale and fine scale mapping, mutant ses was narrowed into a 67 kb interval from CER448792 (2000541 bp) to CER464544 (2067844 bp) crossing over the right of BAC F12K11 to the left of the BAC F4H5 including at most 22 putative genes on the top of chromosome l. In sequence-based map of Arabidopsis genes with Mutant phenotype (SMAGMP) mutant ses was between ATlg06150 (EMB1444) and ATlg08060 (MOM). The SES mapping also showed that developed markers on polymorphism site of CAPC not only were simplified and but worked well. 24 markers from CAPC used in the mapping maybe help Arabidopsis researches with others and the methods related to SES mapping also gave an example of positional cloning.

Expression and localization of recombinant human B2 receptors in the methylotrophic yeast Pichia pastoris.

The human bradykinin B2 receptor (B2R) fused with green fluorescent protein (GFP) at the C-terminal has been expressed in the methylotrophic yeast of Pichia pastoris. In the expression vector, B2R gene was drove under the highly inducible promoter of alcohol oxidase 1 gene of P. pastoris. By fluorescence activated cell sorting (FACS) analysis and western blot analysis, it was proved that B2R recombinant receptor proteins were expressed at high level in the yeast. Further more, the transformants of P. pastoris were monitored with confocal microscopy, a strong green fluorescence was checked out. The recombinant B2R receptor proteins were mainly located on the plasma membrane proved by immunofluorescence microscopy.

Selection of phage-display peptides that bind specifically to the outer coat protein of Rice black streaked dwarf virus.

Several peptides that could bind specifically to the outer coat protein encoded by the S10 gene of Rice black streaked virus (RBSDV) were isolated from a phage-display random 12-mer peptide library. The sequence analysis showed that the amino acid motif (K)K**(*)P, the asterisk denoting any amino acid, might be the core sequence by which the peptides bind to the target protein. The peptide 1 that had a high affinity to RBSDV outer coat protein was synthesized by a chemical method and its fusion protein with glutathione-S-transferase (GST) was produced in an Escherichia coli expression system. The dot and Western blot analyses indicated that RBSDV could be detected with a high sensitivity in crude extracts of diseased plant leaves using a purified GST fusion protein. The circular dichroism (CD) spectroscopy revealed that the synthesized binding peptide but not a nonbinding peptide could bring about a marked change in the conformation of outer coat RBSDV protein. Since the protein functions only when it has correct conformation, the peptides binding specifically to it could possibly disturb the function of the virus outer coat protein and might be used to block the transmission pathway of the virus. Summing up, as these peptides showed a high specificity and sensitivity and diagnostic potential for RBSDV, they may represent the basis of a novel strategy for development of resistance to RBSDV.

Selection and expression of peptides which can change the conformation of p20 protein of rice stripe virus.

Phages with high affinity to the P20 protein of rice stripe virus (RSV) were enriched from phage-displayed random 12-mer peptide library after three rounds of phage display screening. Nine different peptides from the enriched library were selected by enzyme-linked immunosorbent assay (ELISA). The P20 protein from raw extracts of rice leaves infected with RSV could be detected by those 9 peptides displayed on the phage, which suggested that a peptide could be an effective tool for diagnosis of RSV in rice and planthopper. Circular dichroism (CD) spectra of P20 fusion proteins with the binding phages and non-binding phages showed that the conformation of P20 protein was changed after binding to each of the 9 selected 12-mer peptides, which suggested that these peptides might disrupt the function of the P20 protein. Thereafter, those peptides might be used to develop plant resistance and disrupt virus transmission. Three of the 12-mer peptide genes were fused with the glutathione-S-transferase (GST) gene in the vector pGEX 3X. The fusion proteins were obtained from an Escherichia coli expression system and purified. The fusion proteins might have a potential to develop a plant peptide-based resistance to its pathogens and virus diagnosis. It also provided a tool (i) to confirm the inhibition of the function of P20 protein by the fusion peptides in vivo, and (ii) to detect the function of P20 protein and the interaction between the virus and its vector.

Identification of rice black streaked dwarf virus in different cereal crops with dwarfing symptoms in China.

This report describes isolation of virus particles from plants of rice, maize, wheat and sorghum with symptoms of dwarfing collected from two provinces of China, purification of double-stranded RNA (dsRNA) from the virus particles, and synthesis of full-length cDNAs of genome segments 9 (S9) and 10 (S10) by reverse transcription-polymerase chain reaction (RT-PCR). Sequence analysis showed that the S9 sequences of the Chinese isolates and a Japanese rice black-streaked disease virus (RBSDV) isolate were very similar (89.1-89.6% homology at nucleotide level and 92.3-92.9% and 95.8-98.6% homology at amino acid level for ORF1 and ORF2, respectively). Analogical similarity was found also for the S10 sequences of the isolates under comparison: 93.0-95.4% homology at nucleotide level and 96.2-97.0% homology at amino acid level. However, there was a relatively lower similarity for S9 and S10 segments ofthe Chinese isolates and an Italian maize rough dwarf virus (MRDV) isolate. The phylogenetic analysis indicated that the Chinese isolates that infect rice, maize, wheat and sorghum and cause similar symptoms could represent the same virus species, RBSDV.

[Phylogenetic relationships and evolution of yeast-like symbionts of the small brown planthoppers based on partial 18S rDNA sequences].

The yeast-like symbionts are isolated and purified from the small brown planthoppers collected from five areas in China. The 18S rDNA sequences of them are determined. Combined with the sequences of other fungi, we have constructed a comprehensive phylogenetic tree. The results suggested that the taxonomic positions of the YLS are in the class Pyrenomycetes in the subphylum Ascomycotina and the YLS have the closest relationship with H. chrysospermus. The YLS of different areas in China and Japan may belong to different geographical populations of one species. We could also draw a conclusion that there are independent acquisitions of various symbionts in differentiated hosts.

[Production of herbicide-resistant rice with transforming heterogene].

Using pAHC20 (containing Bar gene), pWRG1515 (containing GUS gene and hygromycin phosphotransferase gene), and pCAMBIA3300 RG with Bar gene and snowdrop lectin (GNA) gene as donor DNA, the micro-adventitious shoots and the calli induced from mature embryos of Oryza sativa 87203, Eyi105, Shangnong aromatic glutinous rice as recipients were transformed with particle bombardment and Agrobacterium tumefaciens strain LBA4404 containing pAL4404, respectively. After chosen with phosphinothricin and antibiotic, GUS detection and PCR analysis, The results showed that the foreign genes had been transformed microprojectile-mediated to Oryza sativa Eyi105, the regeneration plants were obtained, and, 5 transgenic calli of Oryza sativa Eyi105 were obtained with Agrobacterium-mediated transformation.

[Western blotting of RStV gene products in rice and insects].

The NS3 and NC protein genes encoded by RNA3 of RStV, the NCP and NSvc4 protein genes encoded by RNA4 were subcloned into the E. coli expression vector pGEX3X to express four groups of fusion protein under IPTG induction. These fusion proteins were used to immunize rabbits to raise antisera. The antisera against the E. coli-expressed proteins were available for probing the presence of the viral gene products in both rice plant and insect hosts. The expected gene products can be probed only in diseased rice plant with NCP antiserum and the corresponding products detected in both plant and RStV particle preparation with NC antiserum. The viral gene products probed by NS3 and NSvc4 antisera were different from the expected ones in size.

Studies on quantitative beta activity in EEG background changes produced by intravenous diazepam in epilepsy.

This article deals with the further systematic investigation of the effects of diazepam on quantitative beta activity in EEG background activity, and the determination of the affecting factors concerned. Diazepam (0.1-0.03 mg/kg) was injected intravenously, the percentage of diazepam-induced changes in beta activity (PDICB) over the 8 sites of scalp EEG recordings was quantitatively analyzed using spectral analysis. The affecting factors concerned were also explored by univariate and multivariate statistical analysis in 84 cases of epilepsy. The results showed that the mean PDICB value in epileptic patients (4.49 +/- 4.51) was significantly lower than that in normal controls (8.03 +/- 5.55). In patients with slowing of EEG background pre-diazepam, AED treatment, long duration of receiving AEDs, refractory epilepsy and brain structural damage, the PDICB value was statistically significantly reduced. The mean PDICB values were lower in the patients with CPS, GTCS, etiologic factors, or paroxysmal discharges on the pre-drug EEG recordings, and the PDICB tended toward decrease with advancing disease duration and seizure frequency; however, these trends did not reach statistical significance. Besides the PDICB values also did not statistically correlate with sex, age and age at seizure onset. In multi-variate analysis with a stepwise regression model, it was found that slowing of EEG background and AED treatment were independently and negatively associated with the decreased PDICB. These results suggest that a reduction of beta activity induced by diazepam, which may involve the whole brain besides the epileptogenic lesion, is significantly related not only to cerebral function impairment of structural changes due to the epileptic process, but also to AED interactions or synergistic effects.

The prognostic significance of diazepam-induced EEG changes in epilepsy: a follow-up study.

Diazepam (0.1-0.3 mg/kg) was injected intravenously, its effect on scalp EEG was evaluated visually and by computer in 84 cases of epilepsy, and long-term follow-up was carried out in an attempt to explore relationships between the EEG changes produced by diazepam and the prognosis in these patients. The average length of follow-up was 3 years (range 2-3.5) in 48 out of 84 cases (57%). The EEG visually responded to diazepam (abolition of abnormal activity with emergence of fast activity) in 33/48 cases. Subsequent follow-up showed that 29 (88%) of these had a good prognosis (seizure-free or a 50% or more reduction in seizures) and 4 others (12%) had a poor result (frequency of seizures increased, unchanged or decreased less than 50%). Fifteen patients had a negative EEG response to the drug, 4 (27%) of whom had a favorable outcome and 11 (73%) an unfavorable result. These results were statistically significant. The percentage of diazepam-induced EEG changes in beta activity (PDICB) was also significantly positively related to the percentage of reduction in seizure frequency in these patients (r = 0.55, p < 0.001). In 79% of patients with PDICB values more than 2, and 30% of those with values less than 2, a good outcome occurred whereas 21% and 70%, respectively, had a poor outcome (p < 0.001). These results showed that the patterns of EEG change induced by diazepam are intimately related to the outcome of epilepsy.

[Effect of (-)-stepholidine on blood pressure and alpha-adrenoceptor agonists-, KCl- and CaCl2-evoked contractions of aortic strips].

Iv (-)-SPD lowered the blood pressure in anesthetized rat, the ED50 value was 5.1 +/- 2.5 mg.kg-1. In the experiments of rat and rabbit aortic strips, (-)-SPD 0.3-100.0 mumol.L-1 inhibited the contraction initiated by clonidine (alpha 2) and phenylephrine (alpha 1) and shifted the dose-response curve to the right parallely without change in maximum response. The inhibitory ratio of (-)-SPD acting on alpha 2/alpha 1 adrenergic receptors was about 7.2, and (-)-SPD thus was predominant inhibition on alpha 2 adrenergic receptors. In the experiment of aortic strips from reserpinized rabbits, the inhibition of (-)-SPD on contraction evoked by clonidine was diminished markedly. The results suggest that (-)-SPD stimulated mainly the alpha 2-adrenergic receptors of presynaptic nerve endings. Moreover (-)-SPD 1 mumol.L-1 inhibited the release of intracellular Ca2+ initiated by NE. (-)-SPD 3-30 mumol.L-1 blocked the voltage-dependent Ca2+ channel.

Transmission electron microscopic study of maize pachytene chromosome 6.

Cytoplasm-free chromosomes are frequently obtained in meiotic chromosome spreads prepared from mildly-fixed maize microsporocytes. These chromosomes are suitable for detailed structural analysis using a published electron microscopic technique. In the electron micrograph, the knobs and heterochromatin regions that have been used for karyotype analyses in the light microscope are clearly visible. Therefore, the electron microscopic map can be easily aligned with the traditional cytological map. In addition to these prominent structural features, numerous electron-dense bands also are observed. To determine whether the bands can be used as markers for the identification of each chromosomal subregion, the banding pattern of chromosome 6 is analyzed. Chromosome 6 is frequently associated with the nucleolus and can be easily recognized. We observed that at the zygotene stage in prophase I, electron-dense regions are detected on each homolog of the synapsing chromosome. During synapsis, the electron-dense regions on both homologs are brought into register to form more conspicuous bands. At the early pachytene stage, the banding pattern is stable and reproducible. Chromosome 6 contains eight dark bands, 19 medium bands and 14 light bands. The bands can be used as intrachromosomal markers for regional assignment of genes in detailed in situ hybridization mapping or cytogenetic studies. As the pachytene stage progresses, condensation of the chromosome bivalents is accompanied by fusion of adjacent bands.