Correlation analysis between serum total IgE and FeNO and idiosyncratic reaction in bronchiolitis.

OBJECTIVE: This article focused on the correlation between the changes of serum total Immunoglobulin E (IgE) and Fractional exhaled Nitric Oxide (FeNO) and idiosyncratic reactions in children with bronchiolitis. METHODS: One hundred children with bronchiolitis and fifty healthy children were enrolled. Serum total IgE and FeNO were assessed, and the diagnostic value for bronchiolitis and the correlation with the severity of bronchiolitis were analyzed. Bronchiolitis children were divided into idiosyncratic + bronchiolitis and non-idiosyncratic + bronchiolitis groups, the relationship between serum total IgE and FeNO and idiosyncratic reaction was determined, and the diagnostic value of serum total IgE and FeNO for idiosyncratic bronchiolitis was examined. RESULTS: FeNO in bronchiolitis children was lower than that in healthy children but there was no significant difference in serum total IgE levels between the two populations. Serum total IgE increased while FeNO decreased with the aggravation of bronchiolitis in bronchiolitis children. The serum total IgE was positively correlated while FeNO was negatively correlated with the severity of bronchiolitis. Serum total IgE was higher in children with idiosyncratic bronchiolitis, but serum total IgE and FeNO were not the risk factors for idiosyncratic bronchiolitis; Area Under the Curve (AUC) of serum total IgE and FeNO for the diagnosis of idiosyncratic bronchiolitis was less than 0.7. CONCLUSION: Serum total IgE and FeNO in children with bronchiolitis are related to disease severity and idiosyncratic reaction. FeNO has a diagnostic value for bronchiolitis, but not for idiosyncratic bronchiolitis.

Facile preparation and dielectric properties of BaTiO3 with different particle sizes and morphologies.

BaTiO3 nanoparticles were prepared by the hydrothermal method, and the effect of 1-(propyl-3-methoxysilyl)-3-methylimidazole chloride on the size of BaTiO3 particles was investigated. The obtained BaTiO3 was characterized by XRD, SEM, TEM, and Raman spectroscopy; and the dielectric properties of BaTiO3 ceramic sheets were tested. The results indicate that the spherical BaTiO3-N prepared without an ionic liquid was in a tetragonal phase with an average particle size of 129 nm. When an ionic liquid was added, the size of the BaTiO3-IL decreased and the degree of agglomeration increased. In addition, with increasing quantity of ionic liquid, the tetragonal-phase content of BaTiO3-IL gradually decreased until complete transformation into cubic phase. The dielectric constant of the BaTiO3-N ceramics was the highest, and the dielectric constant decreased with decreasing BaTiO3 particle size. Moreover, two types of BaTiO3 nanoparticles (bowl- and sea urchin-shaped) were prepared by changing the hydrothermal conditions and additives. The average particle size of the former was 92 nm, the tetragonal-phase content was ca. 90%, and the dielectric constant was large; whereas the sea urchin-shaped BaTiO3 consisted of small particles in the cubic phase, and the dielectric constant was small.

Real-world data in late presentation of HIV infection in Suzhou, China: Results from four consecutive cross-sectional surveys, 2017-2020.

Objectives: This study aimed to examine the real prevalence of late presentation of HIV infection and to identify factors associated with late HIV presentation among patients with newly diagnosed HIV/AIDS in Suzhou, China. Methods: Patients with newly diagnosed HIV/AIDS who registered in national AIDS surveillance system from 2017 to 2020 were included in this study. Late presentation (LP) of HIV infection was defined as HIV diagnosis with a CD4 count < 350 cells/µL or an AIDS-defining event. Multivariable logistic regression analyses were used to identify factors associated with LP. Results: A total of 2,300 patients were enrolled. 1,325 were classified as late presenters, showing a high percentage of 57.6% (95% CI: 54.5-60.7%) and a rise (P = 0.004) over the four-year period. Patients with newly diagnosed HIV/AIDS who were older than 24 years of age (aOR = 1.549, P = 0.001 for 25-39 years; aOR = 2.389, P < 0.001 for 40 years and older), were Suzhou registered residents (aOR = 1.259, P = 0.026), and were from inpatient and outpatient (aOR = 1.935, P < 0.001) were more likely to be late presentation. Conclusions: This study showed a high percentage and a rise of late presentation of HIV infection among patients with newly diagnosed HIV/AIDS in Suzhou, China, which is a challenge for future prevention and control of AIDS. Targeted measures should be urgently implemented to reduce late HIV diagnosis.

Anatomy of the energetic driving force for charge generation in organic solar cells.

Eliminating the excess energetic driving force in organic solar cells leads to a smaller energy loss and higher device performance; hence, it is vital to understand the relation between the interfacial energetics and the photoelectric conversion efficiency. In this study, we systematically investigate 16 combinations of four donor polymers and four acceptors in planar heterojunction. The charge generation efficiency and its electric field dependence correlate with the energy difference between the singlet excited state and the interfacial charge transfer state. The threshold energy difference is 0.2 to 0.3 eV, below which the efficiency starts dropping and the charge generation becomes electric field-dependent. In contrast, the charge generation efficiency does not correlate with the energy difference between the charge transfer and the charge-separated states, indicating that the binding of the charge pairs in the charge transfer state is not the determining factor for the charge generation.

Infection of rhesus macaques with a pool of simian immunodeficiency virus with the envelope genes from acute HIV-1 infections.

BACKGROUND: New simian-human immunodeficiency chimeric viruses with an HIV-1 env (SHIVenv) are critical for studies on HIV pathogenesis, vaccine development, and microbicide testing. Macaques are typically exposed to single CCR5-using SHIVenv which in most instances does not reflect the conditions during acute/early HIV infection (AHI) in humans. Instead of individual and serial testing new SHIV constructs, a pool of SHIVenv_B derived from 16 acute HIV-1 infections were constructed using a novel yeast-based SHIV cloning approach and then used to infect macaques. RESULTS: Even though none of the 16 SHIVenvs contained the recently reported mutations in env genes that could significantly enhance their binding affinity to RhCD4, one SHIVenv (i.e. SHIVenv_B3-PRB926) established infection in macaques exposed to this pool. AHI SHIVenv_B viruses as well as their HIVenv_B counterparts were analyzed for viral protein content, function, and fitness to identify possible difference between SHIVenv_B3-PRB926 and the other 15 SHIVenvs in the pool. All of the constructs produced SHIV or HIV chimeric with wild type levels of capsid (p27 and p24) content, reverse transcriptase (RT) activity, and expressed envelope glycoproteins that could bind to cell receptors CD4/CCR5 and mediate virus entry. HIV-1env_B chimeric viruses were propagated in susceptible cell lines but the 16 SHIVenv_B variants showed only limited replication in macaque peripheral blood mononuclear cells (PBMCs) and 174×CEM.CCR5 cell line. AHI chimeric viruses including HIVenv_B3 showed only minor variations in cell entry efficiency and kinetics as well as replicative fitness in human PBMCs. Reduced number of N-link glycosylation sites and slightly greater CCR5 affinity/avidity was the only distinguishing feature of env_B3 versus other AHI env's in the pool, a feature also observed in the HIV establishing new infections in humans. CONCLUSION: Despite the inability to propagate in primary cells and cell lines, a pool of 16 SHIVenv viruses could establish infection but only one virus, SHIVenv_B3 was isolated in the macaque and then shown to repeatedly infected macaques. This SHIVenv_B3 virus did not show any distinct phenotypic property from the other 15 SHIVenv viruses but did have the fewest N-linked glycosylation sites.

Forced Complementation between Subgenomic RNAs: Does Human Immunodeficiency Type 1 Virus Reverse Transcription Occur in Viral Core, Cytoplasm, or Early Endosome?

Although the process of reverse transcription is well elucidated, it remains unclear if viral core disruption provides a more cellular or viral milieu for HIV-1 reverse transcription. We have devised a method to require mixing of viral cores or core constituents to produce infectious progeny virus by a bipartite subgenomic RNA (sgRNA) system, in which HIV-1 cplt_R/U5/gag/Δpol and nfl sgRNAs are complementary to each other and when together can complete viral reverse transcription. Only the heterodiploid virus containing both the nfl and cplt_R/U5/gag/Δpol sgRNAs can complete reverse transcription and propagate infectious virus upon de novo infection. Dual exposure of U87.CD4.CXCR4 cells with high titers of the homodimeric nfl and cplt_R/U5/gag/Δpol virus particles did not result in productive virus infection. On the other hand, in early endosomes, the HIV-1 sgRNAs released from viral cores can retain function and complete the reverse transcription and result in productive infection. These findings confirm the assumptions that, in natural infection, HIV-1 cores, and likely other retrovirus cores, remain largely intact and do not mix/fuse in the cytoplasm during the reverse transcription process, and circulating cytoplasmic HIV-1 sgRNA (produced through transfection) could not help the complementary sgRNA in the viral core to complement the reverse transcription process.

[Genetic characterization of haemagglutinin of novel H1N1 influenza viruses in Suzhou City].

OBJECTIVE: To investigate the genetic variation and molecular characteristics of HA gene of influenza A (H1N1) virus isolated in Suzhou during from2009 to 2011. METHODS: Viral RNA of 5 Suzhou isolates was isolated and their HA gene were amplified and sequenced by the primers and protocol recommended by WHO, and the sequences together with other sequences downloaded from GenBank were analyzed by several bioinformatics software. RESULTS: Compared with vaccine strain, the average homogeneity of nucleotide and amino acids of 5 Suzhou isolates were between 98.8-99.4% and 98.8-99.4% respectively. All of the 5 strains have 1 amino acids replacement in Sb region and 2 strains have 2 amino acids replacement in Ca region. Strains from and outside Suzhou both showed a trend of clustering by collection year. CONCLUSION: The Suzhou strains are still in stable condition although 1-2 amino acids replacement had happened in antigenic sites.

[The application of molecular differential diagnoses platform in identification of the respiratory pathogens].

AIM: To establish a MDD (molecular differential diagnoses) platform for diagnosing the pathogens that may cause respiratory infection by combination of the advanced Tem-PCR(Target enriched multiplex PCR)with xMAP(multiple analyses profiling), and to evaluate the reliability and further use the platform to test clinic samples. METHODS: 22 throat swab specimen from outpatient patients of respiratory department in First Affiliated Hospital of Suzhou University and 20 respiratory tract lavage fluid specimen from inpatients of respiratory department in Affiliated Children Hospital of Suzhou University were collected, and the nucleic acids of the samples were amplified by Tem-PCR and xMAP. RESULTS: Testing of the the known samples showed that the platform had excellent specificity and sensitivity. Testing of the clinic samples showed that the positive rate of the respiratory tract lavage fluid specimen was 63.6%, higher than that of the throat swab specimen, and that the positive rate of RNA pathogens was higher than that of DNA pathogens. CONCLUSION: A reliable MDD platform for detection of respiratory pathogens has been established with high-throughput detection capacity.

Generation of rat monoclonal antibodies against murine LAIR-1.

The leukocyte-associated Ig-like receptor 1 (LAIR-1), an inhibitory receptor bearing two immunoreceptor tyrosine-based inhibitory motifs (ITIM), is expressed on the majority of peripheral leukocytes, including NK cells, T cells, B cells, monocytes, dendritic cells, granulocytes, and thymocytes and is involved in immunologic regulation and hematopoiesis. Murine LAIR-1 (mLAIR-1) is the homolog molecule of human LAIR-1. Using mLAIR-1-Fc as the immunogen and the technique of rat B lymphocyte hybridoma, we raised three hybridoma cell lines secreting monoclonal antibodies (MAb) to mLAIR-1, designated FMU-mLAIR-1.1, -1.2, and -1.3. Rat immunoglobulin class and subclass of the MAb FMU-mLAIR-1.1 approximately 3 were determined to be IgM, IgG1, and IgM, respectively. All these MAbs can bind the mLAIR-1 in immunocytochemistry and immunohistochemistry. FMU-m LAIR-1.2 worked well not only in Western blot assay but also in recognizing natural LAIR-1 molecules on the surface of P388D1, J774, and WEHI3 cells, and mLAIR-1 cDNA-transfected CHO cells detected by FCM. Thus, successful production of rat anti-murine LAIR-1 monoclonal antibodies provides a new powerful tool for investigation of murine LAIR-1 function in mouse model, both in vitro and in vivo.

[Comparison of the binding affinity of LAIR-1(CD305) and LAIR-2(CD306) with their ligands].

AIM: To compare the binding affinity of LAIR-1 and LAIR-2 with their ligands. METHODS: The ligand-expressing cells were incubated with the LAIR-1 and/or LAIR-2 fusion proteins at different concentration at the same time or in succession. Then the interaction changes and competition were measured by influorescent staining and flow cytometric analysis with the specific mAbs against LAIR-1 or LAIR-2. RESULTS: The membrane ligand for LAIR-1 and LAIR-2 was expressed extensively and ligand recognition was of cross-species. LAIR-2 blocked the interaction of LAIR-1 with its ligand but LAIR-1 didn't block the interaction of LAIR-2 with its ligand. CONCLUSION: LAIR-1 and LAIR-2 probably bind to the same ligand with different affinity, which provides some significant evidence for investigating the molecular mechanisms of LAIR-1 family in modulating immune response.

[Expression and function of CD226 on NK cells activated by Superantigens].

AIM: To study the regulation of superantigen in expression and function of CD226 on NK cells. METHODS: Double fluorescent staining and flow cytometry analysis were employed to detect the expression of CD226 on NK cells from human peripheral blood mononuclear cells (PBMC) stimulated by staphylococcus enterotoxin A (SEA) or SEB. (51)Cr release assay was performed to compare the cytotoxicities of NK cells in different activation models. Laser scanning confocal microscope was used to observe the distribution of CD226 on NK cells activated by superantigens at killing stage. RESULTS: In resting NK cells, the percentage of CD56(+) NK cells and CD56(+) CD226(+) cells were 12.3% and 1.4% respectively, and the cytotoxicity of NK cells against K562 cells was (3.2+/-0.2)% at the ratio of 5:1. After PBMC were stimulated by 0.1 microg/mL SEA and SEB for 1 day, the percentage of CD56(+) NK cells was 13.5% and 14.1% respectively, and the percentage of CD56(+) CD226(+) cells were increased. Interestingly, CD226 was mainly expressed on CD56(dim) NK cells. On day 2 after SEA/SEB stimulation, the proportions of CD56(+) CD226(+) cells among CD56(+) cells were 69.1% and 64.3% in SEA and SEB group respectively. On day 3 after SEA/SEB stimulation, the expression of CD226 on NK cells decreased. Furthermore, the cytotoxicity of NK cells stimulated by SEA or SEB for 3 days against K562 cells were much higher than that of the resting NK cells as well as NK cells cultured without SAg for the same culture time (P<0.05), and reached to the peak at day 2 (82.3%+/-6.9% and 80.6%+/-7.5%, respectively). Additionally, we observed that CD226 molecules were colocalized with LFA-1 at the interface of NK cells which contacted K562 cells. CONCLUSION: The cytotoxicity of NK cells enhanced by SEA or SEB may be correlation with the increased expression of CD226 molecule, and CD226 may be involved in synapse formation of NK cells at killing stage.

[Using flow cytometry to detect peripheral blood eosinophils and their related molecules].

AIM: To set up an accurate and rapid method to detect the phenotype of peripheral blood eosinophils using flow cytometry. METHODS: 10(-4) mol/L of theophylline, 10(-4) of dexamethasone and 10(-8) mol/L of rhIL-5 were added to peripheral blood sampled from normal human subjects. Anti-CD16-PE mAb and FITC-labelled mAbs to eosinophil's molecules were used to perform double labeling staining. Simultaneously, CD16-FL2 was used to install gating so as to accurately locate eosinophils. And then their molecules were examined and analyzed. RESULTS: The eosinophils were accurately located. Theophylline and dexamethasone effectively inhibited activation of eosinophils and shedding of CD62L on eosinophils caused by IL-5. CONCLUSION: Our method can accurately detect eosinophils in peripheral blood samples and their molecules. In addition, only a tiny amount of blood sample is needed and few artificial factors are involved in the detection. Therefore, this method may be an ideal both in basic immunological research and clinical laboratory examination.

[The influence of TLSFJM on the proportion of Th1/Th2-like cell subsets].

AIM: To explore the influence of TLSF(JM) on the proportion of alloantigen activated Th1 and Th2-like cell subsets. METHODS: TLSF(JM) or IL-4 was added to mixed lymocyte reaction(MLR) system. The influence of TLSF(JM) on the proportion of Th1 and Th2-like cell subsets was analyzed by intracellular immunofluorescence staining and FACS. RESULTS: In the TLSF(JM) group, the proportion of IFN-gamma(+) cells differentiated from activated lymphoblast descended from 49.8% to 43.1%, IL-4(+) cells from 75.4% to 43.7% and IL-6(+) cells from 67.8% to 52.6%. The similar tendency was also observed in the unactivated small lymphocytes. CONCLUSION: TLSF(JM) can inhibit both the Th1 and Th2-like cell subsets, but mainly inhibit the Th2-like cell subset, thereby reducing the proportion of Th2-like cell subsets.

Preparation and characterization of a set of monoclonal antibodies to TRAIL and TRAIL receptors DR4, DR5, DcR1, and DcR2.

The tumor necrosis factor-related apoptosis-inducing ligand (TRAIL/Apo 2L) is a novel cytotoxic ligand belonging to TNF superfamily. Among TRAIL receptors, death receptor 4 (DR4) and DR5 containing death domain (DD) in their cytoplasmic region mediate apoptosis-signaling upon TRAIL binding, while decoy receptor 1 (DcR1) and DcR2 with a truncated or non-functional DD play "decoy" role. The interaction of TRAIL and TRAIL receptors plays important roles both in immunoregulation and immune pathogenesis of some diseases. In this study, we raised hybridomas secreting monoclonal antibodies against TRAIL (FMU1.1, 1.2, 1.3), DR4 (FMU1.4), DR5 (FMU1.5, 1.6), DcR1 (FMU1.7) and DcR2 (FMU1.8, 1.9). These MAbs could be used for fluorescent staining and flow cytometry (FCM) analysis as well as immunohistochemistry (IHC). Moreover, FMU1.1, 1.3, 1.4 and 1.5 could be used as coating antibodies paring corresponding polyclonal antibodies to develop sandwich ELISAs to quantitate the soluble TRAIL (sTRAIL), sDR4 or sDR5 in serum samples respectively. In addition, cross-linking of DR4/DR5 by FMU1.4 or FMU1.5 MAbs could induce apoptosis of some DR4/DR5-expressing tumor cells. Thus, this set of monoclonal antibodies against TRAIL or TRAIL receptors may be useful in expression phenotypic and functional study of TRAIL and TRAIL receptors.

Localization of TRAIL/TRAILR in fetal pancreas.

AIM: To observe the localization of TRAIL/TRAIR (DR4, DR5, DcR1, DcR2) in the fetal pancreas. METHODS: Fetal pancreas of 32 weeks of pregnancy were obtained from induced abortions, embedded in paraffin, and 4-microm sections were prepared. The localization of TRAIL/TRAILR in fetal pancreas was investigated by fluorescence immunohistochemical method combined with laser scanning confocal microscopy. RESULTS: TRAIL immunoreactive cells were mainly located on the periphery of the pancreas islets. There were a few DcR1 and DcR2 positive cells whereas there were no immunoreactive cells of DR4 and DR5 in the pancreas islets. In the acini and the ducts of the exocrine pancreas there were no TRAIL/TRAILR immunoreactive cells. CONCLUSION: This study not only describes the distribution of TRAIL/TRAILR in the fetal pancreas, but also provides a morphological basis for deducing the function of TRAIL/TRAILR in pancreas, suggesting that in normal pancreatic islets, the pancreatic cells are resistant towards apoptosis too.

[Establishment and characterization of human alloantigen induced T cell clones].

AIM: To estabilish and analyze stable human T cell clones cultured in-vitro. METHODS: The activated T cells from mixed lymphocyte culture (MLC) were cloned by limiting dilution using irradiated PBMCs as feeder cells. T cell clones were then characterized by immunofluorescence staining and flow cytometry analysis. RESULTS: 16 clones were obtained, of which all but one were alphabetaT cells. Among the 15 alphabetaT cell clones, 9 clones belonged to Th subset, including 3 Th0 and 6 Th2, and 6 clones belonged to Tc subset, including 5 Tc0 and 1 Tc2. CONCLUSION: Fifteen human T cell clones were successfully established, which lays the foundation for study on markers and function of Tc1 and Tc2 subsets.