[The first outbreak of dengue fever and molecular tracing in Puyang, 2019].

Objective: This study retrospectively analyzed an outbreak of dengue fever in Puyang of Henan province in 2019, in order to find the sources of infection. Methods: Dengue virus IgM/IgG and NS1 antigen were tested by colloidal gold method. E gene was amplified by PCR. MegaX was used for sequences alignment to construct evolutionary distance trees. Results: After clinical and laboratory confirmation, there were 81 cases of dengue fever, 17 of which were imported case who were local farmers and worked in Combadia and Thailand, and 64 of which were indigenous cases. The E gene alignment results showed that the pathogen of this epidemic was Vietnamese 1 and highly homologous with the Vietnamese strain. After the local outbreak, dengue virus E gene developed a nucleotide site mutation which can be steadily transmission. Conclusion: The dengue fever outbreak in Puyang was a local outbreak caused by dengue virus type 1, which was associated with imported cases. Gene sequencing showed that the imported pathogen had a relatively stable and transmissible nucleotide mutation after the local epidemic.

Proteomic changes in maize as a response to heavy metal (lead) stress revealed by iTRAQ quantitative proteomics.

Lead (Pb), a heavy metal, has become a crucial pollutant in soil and water, causing not only permanent and irreversible health problems, but also substantial reduction in crop yields. In this study, we conducted proteome analysis of the roots of the non-hyperaccumulator inbred maize line 9782 at four developmental stages (0, 12, 24, and 48 h) under Pb pollution using isobaric tags for relative and absolute quantification technology. A total of 252, 72 and 116 proteins were differentially expressed between M12 (after 12-h Pb treatment) and CK (water-mocked treatment), M24 (after 24-h Pb treatment) and CK, and M48 (after 48-h Pb treatment) and CK, respectively. In addition, 14 differentially expressed proteins were common within each comparison group. Moreover, Cluster of Orthologous Groups enrichment analysis revealed predominance of the proteins involved in posttranslational modification, protein turnover, and chaperones. Additionally, the changes in protein profiles showed a lower concordance with corresponding alterations in transcript levels, indicating important roles for transcriptional and posttranscriptional regulation in the response of maize roots to Pb pollution. Furthermore, enriched functional categories between the successive comparisons showed that the proteins in functional categories of stress, redox, signaling, and transport were highly up-regulated, while those in the functional categories of nucleotide metabolism, amino acid metabolism, RNA, and protein metabolism were down-regulated. This information will help in furthering our understanding of the detailed mechanisms of plant responses to heavy metal stress by combining protein and mRNA profiles.

Identification of a major quantitative trait locus for ear size induced by space flight in sweet corn.

The development of molecular markers has contributed to progress in identifying the gene(s) responsible for favorable variations in maize studies. In this study, quantitative trait locus (QTL) mapping was conducted using simple sequence repeat markers in an F2 sweet corn population from a cross between parental line 1132 and space flight-induced mutant line 751 to identify the loci contributing to an increase in some yield traits. A primary mutated genomic region was located on chromosome 9. In total, 26 QTL were detected for eight yield-related traits and assembled into three clusters on chromosome 9. The largest QTL cluster at bin 9.02/03, primarily contributing to >10% of the phenotypic variation in ear and cob diameters, was likely due to a major QTL. Desired alleles of these QTL were provided by the mutant line 751. The primary action of the major mutant allele was an additive effect. Another mutant locus, which was induced in bin 9.01, increased cob and ear diameters by dominant genetic action.

Management of extensive cervical nodal metastasis in nasopharyngeal carcinoma after radiotherapy: a clinicopathological study.

OBJECTIVES: To evaluate the efficacy of afterloading brachytherapy following radical neck dissection (RND) in the management of extensive cervical lymph node disease in nasopharyngeal carcinoma after radiotherapy; and to examine prospectively prognostic factors and the pathologic behavior of neck disease. PATIENTS: Twenty-seven patients with nasopharyngeal carcinoma who had extensive cervical lymph node metastasis following external radiotherapy were treated with RND. Thirteen of them also underwent afterloading brachytherapy with iridium wire (Ir 192). The RND specimens of the 27 patients were also examined with step serial whole-specimen sectioning. RESULTS: All patients survived and their wounds healed primarily. Pathologic examination revealed 183 tumor-bearing lymph nodes that contained tumors in the neck: level I, 4% (8/183); level II, 53% (96/183); level III, 34% (62/183); level IV, 5% (9/183); and level V, 4% (8/183). Extracapsular tumor extension was seen in 84% of patients. Multivariate analysis identified the number of tumor-bearing lymph nodes detected in the specimens to be the only significant factor that affected control of disease. Although the neck disease in the group of patients who had afterloading brachytherapy was more extensive, the 3-year actuarial tumor control for the groups with and without brachytherapy were 60% and 61%, respectively. CONCLUSIONS: Recurrent cervical lymph nodes after radiotherapy in nasopharyngeal carcinoma are extensive and RND is mandatory for a successful salvage. When the nodal metastasis infiltrate or adhere to surrounding tissue, afterloading brachytherapy with iridium wire can provide satisfactory local tumor control.

[Determination of volatile organic compounds in atmospheric environment].

It is well known that volatile organic compounds (VOCs) are the main photochemical pollutants and ozone precursors of the photochemical smog. Investigation of photochemical pollution in the ambient air must focus on VOCs, but the concentration of VOCs in ambient air is in a very low level (10(-9)-10(-12), volume fraction), so there are difficulties in the determination of VOCs. In this work, based on the TO14A and TO15 methods recommended by the Environmental Protection Agency of United States, an improved method for the determination of fifty-six VOCs, mainly O3 precursors, in atmospheric environment was developed. Operating conditions of VOCs preconcentrator, gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) were optimized. Air sample was first frozen by liquid nitrogen, and then H2O and CO2 were eliminated in the VOCs preconcentrator. The preconcentrated VOCs sample was injected to GC and detected by MS or hydrogen flame ionization detector (FID). The C2-C10 hydrocarbons were separated effectively in capillary columns under the high concentration of CO2. The detection limits were 0.1 microgram.m-3 and the relative standard deviations were in the range from 2.57% to 9.82%. This method has been used for the determination of VOCs in real samples. The results were satisfactory.

In vivo electrophysiologic studies in endothelial nitric oxide synthase (eNOS)-deficient mice.

INTRODUCTION: Endothelial nitric oxide synthase (eNOS) mediates attenuation of the L-type calcium channel and modulates myocyte contractility. Arrhythmogenic afterdepolarizations are seen in vitro in ouabain-treated isolated myocytes from eNOS-deficient mice. The aim of these studies was to characterize the baseline electrophysiologic (EP) phenotype of eNOS-deficient mice and their potential susceptibility to cardiac conduction abnormalities and inducible arrhythmias. METHODS AND RESULTS: Surface ECG and in vivo intracardiac EP studies were performed in 27 mice lacking the eNOS gene and 21 wild-type littermate control mice. Baseline studies were performed in 10 eNOS-deficient mice and 10 wild-type controls. Subsequently, 17 eNOS-deficient mice and 11 wild-type controls were pretreated with digoxin, and ECG and EP testing were repeated. Data analysis revealed no significant differences in ECG intervals or cardiac conduction parameters, except sinus cycle length was higher in eNOS-deficient mice than wild-type mice (P < 0.01). After digoxin pretreatment, 7 of 17 eNOS-deficient mice had inducible ventricular tachycardia and 2 others had frequent ventricular premature beats, compared with only 3 of 11 wild-type mice with inducible ventricular tachycardia. In addition, 2 digoxin-treated eNOS-deficient mice and 1 wild-type mouse had inducible nonsustained atrial fibrillation. CONCLUSION: Mice with a homozygous targeted disruption of the eNOS gene have slower heart rates but no other distinguishable EP characteristics under basal sedated conditions. Partial inhibition of the Na+/K+ ATPase pump with digoxin administration increases ventricular ectopic activity in eNOS-/- mice, a phenotype analogous to afterdepolarizations seen in vitro in this eNOS-deficient mouse model.

The role of fine-needle aspiration cytology in the management of parotid tumors: a critical clinical appraisal.

BACKGROUND: Fine-needle aspiration cytology (FNA) is a well-established tool for investigating many head and neck conditions. It application in parotid tumors is, however, very controversial. This article is aimed at defining the exact role of FNA in the diagnostic workup of patients. METHODS: A retrospective review of a 12-years' experience in a university surgical unit of 186 consecutive patients. Clinical opinion, FNA results, and final pathologic findings were examined. RESULTS: FNA obtained the correct final pathologic condition in 54.3% of cases. It increased the identification of malignancy to 64.5% compared with 26% based solely on clinical signs. Malignant FNA diagnoses (85.7%) and repeatedly inconclusive reports (25.7%) were associated with a higher incidence of malignancy. CONCLUSIONS: Methodological interpretation of FNA results provides useful preoperative information and enables more reliable patient counseling and reduces pathologic surprises. Its enhancement of the preoperative recognition of malignant parotid tumors may alert more stringent attention to the operative margin and hence better tumor clearance.

[Determination of fatty acids in shark cartilage by GC-MS using microwave-assisted digestion and derivatization].

A rapid microwave-assisted digestion and derivatization method for the determination of fatty acids in shark cartilage by GC-MS was developed. The optimum conditions for digestion and derivatization were studied in detail using orthogonal design. The digestion and derivatization were accomplished in 4 minutes at 600 W microwave power using HCl-methanol (1:4, V/V) as digestion and derivatization solvent, and the extraction of the target analytes could be carried out simultaneously. This method is rapid, solvent-saving, and particularly suitable for the rapid determination of fatty acids in solid samples.

Phosphorylation and subcellular translocation of endothelial nitric oxide synthase.

In the vascular endothelium, diverse cell surface receptors are coupled to the Ca2+/calmodulin-dependent activation of nitric oxide (NO) synthase. We now report that, in intact cultured endothelial cells, several drugs and agonists are associated with increased serine phosphorylation of the endothelial NO synthase. We biosynthetically labeled bovine aortic endothelial cells with [32P]orthophosphoric acid, exposed the cells to various drugs and hormones, and then immunoprecipitated the enzyme from cell extracts using a highly specific anti-peptide antibody. The marked endothelial NO synthase phosphorylation induced by bradykinin is maximal only after 5 min of agonist exposure and is stable for at least 20 min. Basal and agonist-induced phosphorylation of the NO synthase in endothelial cells is completely inhibited by the calmodulin antagonist compound W-7. We prepared subcellular fractions of endothelial cells that had been biosynthetically labeled with [35S]methionine or [32P]orthophosphoric acid and immunoprecipitated the endothelial NO synthase from untreated (basal) and bradykinin-treated cells. In the basal state, [35S]methionine-labeled endothelial NO synthase is associated primarily with the particulate cellular fraction, but the phosphorylated enzyme is primarily cytosolic. Following exposure to bradykinin, a substantial fraction of the [35S]methionine-labeled NO synthase is now found in the cytosolic fraction, associated with a marked increase in the level of cytosolic enzyme phosphorylation. We propose that agonist-induced phosphorylation of NO synthase is associated with translocation of the enzyme from membrane to cytosol and may thereby regulate the biological effects of endothelial NO synthesis in situ.

Endothelial nitric oxide synthase: molecular cloning and characterization of a distinct constitutive enzyme isoform.

Nitric oxide (NO) is a ubiquitous intercellular messenger molecule synthesized from the amino acid L-arginine by NO synthases in diverse cells and tissues. NO is synthesized in vascular endothelial cells and appears to play an important role in the control of blood pressure and platelet aggregation. A detailed understanding of the regulation of NO synthesis by endothelial cells has been hampered by the lack of molecular clones for endothelial NO synthase; the isolation and characterization of such clones is reported herein. The constitutive NO synthases present in endothelial cells and in brain share common biochemical and pharmacologic features. We purified NO synthase from bovine brain and determined the amino acid sequence of several tryptic peptides. The sequence of the bovine brain peptides is nearly identical to the deduced amino acid sequence previously determined for the rat brain NO synthase. These sequence data were utilized to design PCR-generated NO synthase cDNA probes, which were used to isolate clones encoding NO synthase from a bovine aortic endothelial cell (BAEC) cDNA library. A full-length NO synthase cDNA clone was isolated, representing a protein of 1205 amino acids with a molecular mass of 133 kDa; transfection of this clone in a heterologous expression system demonstrated the expected enzymatic activity. The deduced amino acid sequence of the BAEC NO synthase cDNA differs at numerous residues from the sequence determined for the purified bovine brain protein and shows 50-60% sequence identity with recently isolated molecular clones for murine macrophage and rat brain NO synthase isoforms. Bovine genomic Southern blots probed with bovine brain and BAEC NO synthase cDNA probes identify distinct bands, indicating that these cDNAs are the products of different genes. Prolonged treatment of BAECs with the cytokine tumor necrosis factor alpha, which we have previously shown to result in a marked increase in NO synthase activity, is associated with a decrease in the abundance of the 4.8-kilobase BAEC NO synthase transcript. The increase in BAEC NO synthase activity induced by tumor necrosis factor alpha is thus likely to involve posttranscriptional mechanisms or the induction of a distinct endothelial NO synthase isoform.