[Analysis of hemagglutinin-neuraminidase gene characteristics of human parainfluenza virus type 3 among children with acute respiratory tract infection in Qingdao city].

The purpose was to discuss the infection status of human parainfluenza virus type 3 (HPIV-3) in children with acute respiratory tract infection(ARTI) in Qingdao, Shandong province, and to analyze the gene characteristics of HPIV-3 hemagglutinin-neuraminidase protein (HN). This study was a cross-sectional study. A total of 1 674 throat swab samples were collected randomly from children with ARTI, in the three hospitals (Qingdao Women and Children's Hospital, West Coast Branch of Affiliated Hospital of Qingdao University, Laoshan Branch of Affiliated Hospital of Qingdao University) from January 2018 to December 2019. Multiplex real-time fluorescence RT-PCR was performed to screen HPIV-3 positive specimens. For HPIV-3 positive specimens, nested PCR was used to amplify the full-length HN gene of HPIV-3. The HN gene was sequenced and compared with the representative strains of HPIV-3 in GenBank, and the phylogenetic tree was established. As results, this study collected 1 674 samples, in which there were 90 HPIV-3 positive samples showed and the detection rate was 5.37%. Among positive specimens, the number of samples from children under 6 years old was 88, accounting for 97.78%. HPIV-3 positive cases were mainly distributed in spring and summer. The full-length sequences of 44 HPIV-3 HN genes were obtained by nested PCR method. Sequence alignment and evolutionary analysis showed that the HPIV-3HN gene belonged to the C3a and C3b branches of C3 genotype, with 30 strains of subtype C3a and 14 strains of subtype C3b. The nucleotide and amino acid homology of the amplified 44 strains of the HPIV-3 HN gene in Qingdao were 97.0%-100.0% and 98.5%-100.0%, respectively. In conclusion, from 2018 to 2019, the C3a and C3b branches of HPIV-3 C3 genotype were circulating prevalent in Qingdao, Shandong province. HN gene variation rate was low, but showed certain regional characteristics in evolution.

[Etiology and clinical analysis of central nervous system infection caused by Coxsackievirus B5 in severe hand, foot and mouth disease in Qingdao City, 2013-2014].

Objective: To illuminate the gene characteristics and clinical characterization of Coxsackievirus B5 (CV-B5) strains isolated from patients with sevre hand, foot and mouth disease (HFMD) in Qingdao city. Methods: A total of 1 844 patients of HFMD were consecutively admitted to Qingdao Women and Children's Hospital from 2013 to 2014. Information of the study population described above was collected retrospectively. The samples were collected from at least 1 site (throat swab, cerebrospinal fluid), which viral nucleic acid extracted and the entire VP1 gene sequences of CV-B5 isolates were amplified and sequenced, then the homology and phylogeny analysis were conducted by MEGA7.0. The prototype Faulkner strain and other VP1 amino acid sequences were derived from the GenBank database. Results: A total of 8 CV-B5 positive cases were obtained, including 4 males and 4 females; 6 severe hospitalized cases and 2 outpatients. The age of 6 hospitalized patients ranged from 3 to 48 months, with a median of 26 months. For the six inpatients, fever, convulsions vomiting, diarrhea and rash were the main clinical manifestation, and all combined with viral encephalitis. Compared with the prototype strain Faulkner, in the VP1 region,the nucleotide and the amino acid homologies was 77.3%-78.8% and 95.5%-97.0% respectively. Five out of the six severe cases with substitution of serine (S) to asparagine (N) at amino acid site 95 in the VP1 region. The sequences of 8 CV-B5 strains were classified into genogroup D. Conclusion: Hand, foot and mouth disease associated with CV-B5 virus infection can result in nervous system involvement and the main complication was viral encephalitis. The CV-B5 strains associated with severe hand, foot and mouth disease had high nucleotide homology and present a certain regional aggregation.

Autophagy protein p62/SQSTM1 is involved in HAMLET-induced cell death by modulating apotosis in U87MG cells.

HAMLET is a complex of oleic acids and decalcified α-lactalbumin that was discovered to selectively kill tumor cells both in vitro and in vivo. Autophagy is an important cellular process involved in drug-induced cell death of glioma cells. We treated U87MG human glioma cells with HAMLET and found that the cell viability was significantly decreased and accompanied with the activation of autophagy. Interestingly, we observed an increase in p62/SQSTM1, an important substrate of autophagosome enzymes, at the protein level upon HAMLET treatment for short periods. To better understand the functionality of autophagy and p62/SQSTM1 in HAMLET-induced cell death, we modulated the level of autophagy or p62/SQSTM1 with biochemical or genetic methods. The results showed that inhibition of autophagy aggravated HAMLET-induced cell death, whereas activation of authophagy attenuated this process. Meanwhile, we found that overexpression of wild-type p62/SQSTM1 was able to activate caspase-8, and then promote HAMLET-induced apoptosis, whereas knockdown of p62/SQSTM1 manifested the opposite effect. We further demonstrated that the function of p62/SQSTM1 following HAMLET treatment required its C-terminus UBA domain. Our results indicated that in addition to being a marker of autophagy activation in HAMLET-treated glioma cells, p62/SQSTM1 could also function as an important mediator for the activation of caspase-8-dependent cell death.

Large-area fabrication of periodic Fe nanorings with controllable aspect ratios in porous alumina templates.

Highly uniform Fe nanoring arrays in porous anodic alumina templates are fabricated by physical vapour deposition and grazing ion milling techniques. The nanorings have aspect ratios ranging from 0.8 to 4, depending on the deposition conditions. The outer diameter of the individual nanorings, and the area density and distribution patterns are completely determined by the template used. Selected-area electron diffraction reveals that these nanorings have a polycrystalline microstructure. The nanoring fabrication method demonstrated here can be extended to other materials.

[Protective effect of ischemic preconditioning on ischemia-reperfusion injury].

OBJECTIVE: To study the protective effect of ischemic preconditioning(IP) on ischemia-reperfusion injury of limb. METHODS: One to three times of short-term ischemia was applicated before prolonged ischemia, a series of biological examinations, including creatine phosphate kinase(CPK), glutamic-oxalacetic transaminase(GOT), lactic dehydrogenase(LDH), malondiadehyde(MDA) and superoxide dismutase(SOD) were carried out in each group. RESULTS: Compared with the control, release of skeletal muscle intracellular enzyme(CPK, GOT and LDH) decreased in experiment groups after preconditioning had been performed. The serum concentration of MDA was lower, activity of SOD was higher, and pathological lesion was less in preconditioning groups than that in non-preconditioning group. Different frequency of preconditioning led to different results. CONCLUSIONS: Preconditioning has protective effect on ischemia-reperfusion injury of limbs, and the effect of IP is correlated with the number of preconditioning cycles.

Interstitial lung macrophages interact with dendritic cells to present antigenic peptides derived from particulate antigens to T cells.

When the protective structural and functional barriers of the lung are breached, immune responses must be generated in order to contain invading micro-organisms. This requires the presence of accessory cells capable of phagocytosing and presenting immunogenic peptides to either naive or sensitized T cells. In contrast to dendritic cells (DC) present in the airway epithelium, those within the lung parenchyma do not readily engulf particulates and, therefore, other mechanisms must account for their apparent ability to present immunogenic peptides derived from micro-organisms. The purpose of the present study was to determine the extent to which interstitial macrophages (IM) interact with lung DC to process and present antigenic peptides, derived from particulate, heat-killed Listeria monocytogenes (HKL), to HKL-immune T cells. Results show that highly purified Ia- lung IM avidly phagocytose fluorescent-labelled HKL, but they do not present antigen to primed T cells. Their ability to present antigen is only modestly increased following interferon-gamma (IFN-gamma) stimulation. Conversely, mature DC isolated from the lung interstitium do not phagocytose fluorescent-labelled HKL. In antigen presentation assays, however, addition of 10% (2.5 x 10(3)/ml) Ia- IM to DC and HKL results in a two- to threefold increase in antigen presentation by DC to HKL-immune T cells. Conditioned medium (CM), generated by 2.5 x 10(4)/ml IM induced to phagocytose HKL, when administered to DC and HKL-sensitized T cells without added intact HKL, resulted in brisk mitogenesis, a response that did not occur in T cells sensitized to an irrelevant antigen. Conditioned medium derived from larger numbers of IM was inhibitory. When IM phagocytosed inert polystyrene beads, the resulting CM induced modest T-cell mitogenesis, suggesting that small amounts of cytokines were released. The results indicate that in small numbers, IM augment DC function, in part, by the release of antigenic peptides which are then presented by DC to T cells. When present in numbers greater than 50% of DC, however, they inhibit DC function, probably due to the release of soluble inhibitors.

Ontogeny of Ia+ accessory cells in fetal and newborn rat lung.

In the adult mammalian lung, Ia+ dendritic cells (DC) constitute a significant population of immunologically potent accessory cells that are important in the regulation of immune responses to inhaled antigens. The newborn, in most species, displays an increased susceptibility to sensitization by inhaled antigens; whether an immaturity of pulmonary accessory cells is involved has not been determined. In the present study, the ontogeny and function of these cells were examined in fetal and newborn rats. Cells identified as DC in fetal and newborn rat lungs were Ia+, C11b+/-, OX41-, OX43-, W3/13-, W3/25-, and OX8-. They were characterized ultrastructurally by an eccentric, lobulated nucleus, a paucity of lysosomes, delicate cytoplasmic processes, and abundant membrane-associated Ia. Ia+ DC were first detected within the pulmonary mesenchyme at day 15 and by day 17 of gestation they were also present within the epithelium lining airways. The appearance of Ia+ DC preceded the migration of either T4 or T8 subclasses of T cells to the lung, the latter becoming significant only after birth, when the newborn was exposed to environmental antigens. In none of the fetal or newborn animals was Ia detected on alveolar type II cells. The accessory cell function of rat pulmonary DC, isolated from fetuses at 20 and 21 days of gestation and from newborns, was tested by an autologous mixed leukocyte reaction. At 20 and 21 days of gestation, pulmonary DC were 40 and 60% as effective, respectively, in stimulating cell proliferation in purified autologous adult splenic T cells as those isolated from adults.(ABSTRACT TRUNCATED AT 250 WORDS)

Intraepithelial airway dendritic cells: a distinct subset of pulmonary dendritic cells obtained by microdissection.

Dendritic cells (DC), in general, and pulmonary DC, in particular, are a heterogeneous population of cells, their phenotype and function being dependent on their anatomic location, their state of activation, and the regulatory effect of locally secreted cytokines. Using a novel microdissection technique, the epithelium from the trachea and entire airway system was harvested, and the contained DC isolated at greater than 90% purity. The phenotype and function of these airway DC (ADC) was compared to DC isolated, at greater than 90% purity, from the parenchyma of the same lung. In contrast to lung DC (LDC), ADC did not express intercellular adhesion molecule 1 (ICAM-1) in situ, the amount of immune associated antigen (Ia) expressed was less (as determined by immunoperoxidase staining and immunopanning), and greater than 50% of ADC displayed Fc receptors (FcR). The majority of LDC were ICAM-1+, less than 5% expressed FcR, and all were intensely Ia+. Airway DC were most numerous in tracheal epithelium, but they were also present in small numbers in the epithelium of the most distal airways. Their numbers increased in all segments of the tracheobronchial epithelium in response to the administration of IFN-gamma. ADC were consistently more effective than LDC in presenting soluble (hen egg lysozyme) and particulate (heat-killed Listeria monocytogenes) antigens to antigen-sensitized T cells. By contrast, LDC were significantly more efficient in stimulating the proliferation of nonsensitized T cells in an autologous mixed leukocyte reaction. These data suggest that in normal animals, intraepithelial DC of airways share many attributes with Langerhans cells of the skin. Interstitial LDC, by contrast, reside in an environment where they may be exposed to a different set of regulatory factors and where they have progressed to a more advanced stage of differentiation than ADC. Both groups of DC are, however, heterogeneous, reflecting the continuous turnover that these cells undergo in the lung.

A new procedure for labeling alkylbenzenes with [18F]fluoride.

A new procedure for labeling alkylbenzenes with no-carrier-added (nca) [18F]fluoride is reported. This will allow the use of [18F]-for-nitro aromatic nucleophilic displacement reaction for labeling aromatic compounds with no activating groups on the benzene ring. The new procedure involves (A) the [18F]-for-nitro displacement reaction on nitrophenones, and (B) the reduction of [18F]fluorophenones with triethylsilane and trifluoroacetic acid to alkylfluorobenzenes. The desired 18F-labeled alkylbenzenes were prepared in a synthesis time of 1 h with a radiochemical yield of 20% at end-of-synthesis. The procedure has been successfully applied to the synthesis of 18F-labeled alkylating agents, such as 4-[18F]fluorophenethyl bromide, 4, and 4-[18F]fluorophenbutyl chloride, 5. Using the reaction of piperidine and 4 as a model, the potential use of phenethylbromide 4 for labeling biologically important amines was examined. Initial results indicated that the desired alkylated piperidine was formed in low yields (less than 5%) due to the conversion of halide 4 to [18F]fluorostyrene (greater than 85%) under basic conditions. The new procedure provides an easy method of labeling alkylbenzenes with fluorine-18.