Objective Base on the neighborhood activation model(NAM), to develop the Mandarin monosyllable lexical neighborhood test(M-LNT-monosyllable), which is expected to be useful for evaluating speech perception performance in children with hearing loss. Methods Test items were based on the lexical characteristics of word frequency and neighborhood density which addressed in the neighborhood activation model (NAM). M-LNT -monosyllable consisted of two parts: Lexically "easy" words with high word frequency, which were low phonemically similar to the target word and lexically "hard" words with low word frequency, which were high phonemically similar to the target word. 34 children of 3～5 year old with normal hearing were choosed as subjects to verify easy word and hard word lists. Results 1 979 words for children contained 487 easy words and 419 hard words. Three easy word lists and three hard word lists were developed to estimate the performance of word recognition among normal- hearing children. There were no differences among scores of three easy words lists(P＞0.05), and no difference among scores of three hard words lists(P＞0.05). But there were significant differences between scores of easy and hard words lists(P＜0.01). Conclusion The development of the lexicon was affected by the lexical characteristics. Normal-hearing children with some lexical techniques were affected by the lexical characteristics when they recognized the spoken words, but children with less lexical technique didn't show the same result because the recognition was processed on the phonetic level.

OBJECTIVETo observe whether dendritic cells (DCs) transfected with recombinant adenovirus Ad5F35-LMP2 induces LMP2 specific immunity mediated by cytotoxic T lymphocytes in vitro.

METHODSDendritic cells have been generated in vitro, and cocultured with autologous T cell after the DCs were infected with Ad5F35-LMP2, then the proliferation of the induced T cells and their cytotoxic activity against CNE-2 tumor cells which express EBV-LMP2 protein on membrane were assessed by MTT method.

RESULTSThe dendritic cells could be transfected with Ad5F35-LMP2 and the CTL activated by Ad5F35-LMP2-DC could effectively suppress the proliferation of CNE-2 cells compared with control groups.

CONCLUSIONThe dendritic cells transfected with recombinant adenovirus Ad5F35-LMP2 showed cytotoxicity effect by activating T lymphocytes.

Adenoviridae ; genetics ; Cell Line, Tumor ; Cells, Cultured ; Dendritic Cells ; immunology ; virology ; Epstein-Barr Virus Infections ; immunology ; virology ; Genetic Vectors ; genetics ; Humans ; Immunity, Cellular ; T-Lymphocytes, Cytotoxic ; immunology ; virology ; Viral Matrix Proteins ; genetics ; immunology

BACKGROUNDEpstein-Barr virus (EBV) is a herpesvirus commonly associated with several malignant diseases including nasopharyngeal carcinoma (NPC), which is a common cancer in Southeastern Asia. Previous studies showed that plasma levels of EBV-DNA might be a sensitive and reliable biomarker for the diagnosis, staging and evaluating of therapy for NPC. There are a few analyses of the levels of EBV-latent membrane protein 2 (LMP2)-specific cytotoxic T-lymphocytes (CTLs) in patients with NPC. This study was conducted to investigate the levels of EBV-LMP2-specific CTLs, EBV-DNA load and the level of CD4(+)CD25(+) T cells in such patients.

METHODSFrom February 2006 to April 2006, 62 patients with NPC, 40 healthy virus carriers positive for EBV viral capsid antigen (EBV-IgA-VCA) and 40 controls were enrolled in the study. We used a highly sensitive ELISPOT assay, real-time polymerase chain reaction (PCR) and flow cytometry to measure the EBV-LMP2-specific CTL response, the EBV DNA load and the level of CD4(+)CD25(+) T cells, respectively.

RESULTSThe EBV-LMP2-specific CTL responses of the samples from the control, healthy virus carriers and patients with NPC were significantly different from the LMP2 epitopes, with the control and healthy virus carrier samples displaying a stronger response in three cases. There were significant differences in EBV DNA load in serum between NPC and the healthy groups; patients with NPC at stages III or IV had significantly higher viral loads compared with those at stages I or II. A significantly higher percentage of CD4(+)CD25(+) T lymphocytes were detected in the patients, compared with healthy virus carriers and healthy controls. Moreover, patients with advanced stages of NPC (III and IV) had significantly higher percentages than the patients with early stages (I and II).

CONCLUSIONSPatients with NPC are frequently unable to establish or maintain sufficient immunosurveillance to control proliferating B cells harboring EBV and to destroy the tumor cells that express immunodominant LMP2 proteins. Controlling the activity of CD4(+)CD25(+) T cells and elevating CD8(+) cells specific for LMP2 epitopes could be an effective immunotherapy for patients with NPC.

Adult ; Antigens, Viral ; immunology ; CD4-Positive T-Lymphocytes ; immunology ; Capsid Proteins ; immunology ; DNA, Viral ; genetics ; Epstein-Barr Virus Infections ; immunology ; virology ; Female ; Flow Cytometry ; Humans ; Immunoglobulin A ; immunology ; Interleukin-2 Receptor alpha Subunit ; immunology ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; immunology ; virology ; Polymerase Chain Reaction ; T-Lymphocytes, Cytotoxic ; immunology ; Viral Matrix Proteins ; immunology

OBJECTIVETo study the effect of liposomal transfection of antisense oligonucleotide (ASON) on the erythroid cell alpha-globin gene in the patients with severe beta-thalassemia, and provide a new idea for beta-thalassemia gene therapy.

METHODSA highly effective ASON targeting alpha-globin gene was transfected into severe beta-thalassemic erythroid cells cultured in vitro by liposomal at an optimal concentration. The expression level of alpha, beta, gamma-globin gene, the level of hemoglobin, and the excess alpha-globin chains precipitates in ASON group and control group were carefully analyzed by quantitative real-time PCR(Q-RT-PCR), high performance liquid chromatography (HPLC), and electron microscope, respectively.

RESULTSThe mRNA expression of alpha-globin gene was significantly lower in ASON group (9.04 +/- 0.29) than in control group (24.23 +/- 0.29) (P<0.01). Simultaneously, the disequilibrium between alpha- and beta-, gamma-globin gene expression was partly modified by ASON, the ratios of ASON group and control group being 0.79 +/- 0.02 and 2.26 +/- 0.06 respectively (P<0.01). HPLC demonstrated that the levels of HbA2 and HbF increased with downregulation of alpha-globin gene in beta-thalassemic erythroid cells, particularly HbF. The precipitates of alpha-globin chains in ASON group were lessened under electron microscope, particularly in early erythroblast while no change in the control group.

CONCLUSIONThe high effective ASON contributes to inhibit the alpha-globin gene expression of severe beta-thalassemic erythroid cells, partly modify the disequilibrium between alpha-, beta- and gamma-globin gene expression and obviously reduce the precipitates of alpha-globin chains in erythroid cells. It might provide a new idea for gene therapy of beta-thalassemia.

Cells, Cultured ; Child ; Genetic Therapy ; Humans ; Liposomes ; Oligonucleotides, Antisense ; genetics ; Transfection ; alpha-Globins ; genetics ; metabolism ; beta-Globins ; metabolism ; beta-Thalassemia ; genetics ; metabolism ; therapy ; gamma-Globins ; metabolism

OBJECTIVETo explore GAGA-like element binding protein in human cells.

METHODSYeast one-hybrid system was used to screen the GAGA-like element binding proteins in HTLV-1 transformed Jurkat cell cDNA fusion library. Total RNA extracted from Jurkat cells was first labeled by reverse transcription, and was taken as cDNA probe to hybridize with the candidate positive clones.

RESULTS9 positive clones were obtained, and 6 out of the 9 clones were positively hybridized with the cDNA probe.

CONCLUSIONS6 candidate clones encoding for GAGA-like element binding proteins were obtained from Jurkat cells for further investigation.

Carrier Proteins ; biosynthesis ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; DNA-Binding Proteins ; Drosophila Proteins ; HSP90 Heat-Shock Proteins ; biosynthesis ; genetics ; Homeodomain Proteins ; genetics ; Human T-lymphotropic virus 1 ; genetics ; Humans ; Jurkat Cells ; Leukemia, T-Cell ; metabolism ; pathology ; Transcription Factors ; genetics ; Two-Hybrid System Techniques

BACKGROUNDInterleukin 16 (IL-16) can be detected by ELISA in pleural effusion (PE) and its concentration is higher than in serum. This study investigated the cellular sources of IL-16 in PE.

METHODSThe samples of PE were collected from 34 patients who were newly diagnosed having PE in the pleural cavity. We performed cell culture to purify the pleural mesothelial cells (PMC), Wright staining to count the purity and immunocytochemical stain to identify the cultured cells. The intracellular IL-16 expression was detected by flow cytometry (FCM). The different cells in PE were first separated by magnetic cell sorting (MCAS) then the separated cells were cultured in RPMI1640 with 10% fetal calf serum (FCS). We extracted the supernatant and detected IL-16 concentration by ELISA. The IL-16 protein was detected by immunohistochemistry and double immunofluorescence staining.

RESULTSThe percentages of cells which secreted IL-16 were: CD3(+)CD8(-) cells ((74.27 ± 15.56)%, n = 34); CD3(+)CD8(+) cells ((69.86 ± 18.55)%, n = 34); CD19(+) cells ((45.30 ± 18.77)%, n = 15); CD14(+) cells ((16.91 ± 16.69)%, n = 15); and PMC ((2.05 ± 1.85)%, n = 7). The concentrations of IL-16 in the supernatant from cultured cells were: CD4(+) cells ((102.50 ± 42.51) ng/L, n = 5); CD8(+) cells ((92.58 ± 18.34) ng/L, n = 5); CD19(+) cells ((79.85 ± 5.62) ng/L, n = 5); CD14(+) cells ((58.51 ± 25.38) ng/L, n = 5); and PMC ((18.14 ± 8.37) ng/L, n = 5). In lymphocytes, monocytes/macrophages and PMC, we could observe the cells that expressed IL-16 protein. In paraffin-embedded sections, we also could observe by immunohistochemistry the CD4(+)IL-16(+) cells, CD8(+)IL-16(+) cells, CD19(+)IL-16(+) cells, and CD14(+)IL-16(+) cells.

CONCLUSIONSIL-16 in PE is mainly secreted by T lymphocytes, including CD3(+)CD8(-) cells and CD3(+)CD8(+) cells. CD19(+) cells and CD14(+) cells can also secrete IL-16, but the percentage of PMC that can secrete IL-16 is very low.

Adult ; Aged ; Aged, 80 and over ; Antigens, CD19 ; metabolism ; CD4-Positive T-Lymphocytes ; metabolism ; CD8-Positive T-Lymphocytes ; metabolism ; Cells, Cultured ; Centrifugation, Density Gradient ; Enzyme-Linked Immunosorbent Assay ; Female ; Flow Cytometry ; Humans ; Immunohistochemistry ; Interleukin-16 ; metabolism ; Lipopolysaccharide Receptors ; metabolism ; Male ; Middle Aged ; Pleural Effusion, Malignant ; metabolism ; T-Lymphocytes ; metabolism ; Young Adult

OBJECTIVETo observe the specific cellular and humoral immune responses after immunization with recombinant adenovirus Ad5F35-LMP2 in rhesus monkeys.

METHODSSixteen rhesuses were immunized with Ad5F35-LMP2 through intra-muscular injection in three groups: high dosage group (1.5 x 10(10) TCID(50)/rhesus), medium dosage group (1.5 x 10(9)TCID(50)/rhesus), low dosage group (1.5 x 10(8)TCID50/rhesus) and the last group was control (PBS 4 ml/rhesus). They were totally immunized three times at intervals of one month. The EBV-LMP2 specific cellular immune responses were tested during the 0, 4, 8, 12 weeks by Elispot after immunization respectively. And the titers of anti-LMP2 antibody were tested by EIA at the same time.

RESULTSEBV-LMP2 specific cellular and humoral immune responses which were induced by recombinant adenovirus Ad5F35-LMP2 can be found in all the three dosage groups. The potency of immune responses was related with the dosage of immunization. Higher dosage elicited more potent immune response.

CONCLUSIONThe recombinant adenovirus Ad5F35-LMP2 could elicit LMP2 specific cellular and humoral immune responses in rhesus.

Adenoviruses, Human ; genetics ; Animals ; Cell Differentiation ; Herpesvirus 4, Human ; genetics ; Immunity, Cellular ; immunology ; Immunization ; methods ; Macaca mulatta ; Recombinant Fusion Proteins ; genetics ; immunology ; Viral Matrix Proteins ; genetics ; immunology

BACKGROUND/OBJECTIVES: Tibetan tea is a kind of dark tea, due to the inherent complexity of natural products, the chemical composition and beneficial effects of Tibetan tea are not fully understood. The objective of this study was to unravel the composition of Tibetan tea using knowledge-guided multilayer network (KGMN) techniques and explore its potential antioxidant and hypolipidemic mechanisms in mice.MATERIALS/METHODS: The C57BL/6J mice were continuously gavaged with Tibetan tea extract (T group), green tea extract (G group) and ddH 2 O (H group) for 15 days. The activity of total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) in mice was detected.Transcriptome sequencing technology was used to investigate the molecular mechanisms underlying the antioxidant and lipid-lowering effects of Tibetan tea in mice. Furthermore, the expression levels of liver antioxidant and lipid metabolism related genes in various groups were detected by the real-time quantitative polymerase chain reaction (qPCR) method. RESULTS: The results showed that a total of 42 flavonoids are provisionally annotated in Tibetan tea using KGMN strategies. Tibetan tea significantly reduced body weight gain and increased T-AOC and SOD activities in mice compared with the H group. Based on the results of transcriptome and qPCR, it was confirmed that Tibetan tea could play a key role in antioxidant and lipid lowering by regulating oxidative stress and lipid metabolism related pathways such as insulin resistance, P53 signaling pathway, insulin signaling pathway, fatty acid elongation and fatty acid metabolism. CONCLUSIONS This study was the first to use computational tools to deeply explore the composition of Tibetan tea and revealed its potential antioxidant and hypolipidemic mechanisms, and it provides new insights into the composition and bioactivity of Tibetan tea.

Objective:To investigate the risk factors for bleeding and thrombosis during extracorporeal membrane oxygenation (ECMO) therapy in critically ill patients and determine the best predictors of coagulation-related complications.Methods:A retrospective analysis was performed on patients who received ECMO for respiratory or circulatory failure at Xiangya Hospital of Central South University from January 2020 to December 2022. The outcome was whether bleeding or thrombosis occurred from 24 h after ECMO insertion to before weaning. The differences in demographic characteristics, weaning conditions, prognosis, routine blood tests, organ function, coagulation and blood product transfusion of each group were compared. Logistic regression analysis was used to evaluate the risk factors for bleeding and thrombosis, and ROC curve evaluation was used to assess their capacity to predict complications.Results:A total of 61 patients with ECMO were enrolled, with 21 cases of bleeding and 14 cases of thrombosis during ECMO. Compared with the nonbleeding group, the activated partial thromboplastin time, thromboplastin time (TT), and transfusions of frozen plasma and red blood cells were higher in the bleeding group (all P<0.05). Compared with the nonthrombotic group, the increase in body weight, D-dimer (DD), fibrinogen degradation product (FDP), and improvement of arterial oxygen partial pressure (ΔPO 2) within 24 h were significantly higher in the thrombotic group (all P<0.05). Logistic regression analysis revealed that TT ( OR=1.039, 95% CI: 1.006~1.072, P=0.018) and frozen plasma transfusion volume ( OR=1.046, 95% CI: 1.010-1.083, P=0.012) were risk factors for bleeding events. FDP ( OR=1.030, 95% CI: 1.009-1.051, P=0.005), DD ( OR=1.181, 95% CI: 1.044-1.336, P=0.008), and ΔPO 2 ( OR=1.007, 95% CI: 1.002-1.012, P=0.006) were risk factors for thrombosis. According to ROC curve analysis, the AUCs of TT, frozen plasma transfusion, and combined indexes for predicting bleeding were 0.712, 0.690, and 0.816, respectively. The combined indices had a cut-off value of 0.273, a sensitivity of 75.61%, and a specificity of 80.00%. The AUCs of FDP, DD, ΔPO 2, and combined FDP with ΔPO 2 for predicting thrombosis were 0.778, 0.748, 0.786, and 0.868, respectively. The cut-off value of the combined index was 0.157, the sensitivity was 68.09%, and the specificity was 92.86%. Conclusions:TT combined with frozen plasma transfusion volume predicted bleeding optimally, while FDP plus ΔPO 2 predicted thrombotic events better during ECMO treatment in critically ill patients.

OBJECTIVETo study the role of a BTB/POZ domain protein in the expression of hsp90alpha gene.

METHODSThe eukaryotic expression plasmids of sense- and antisense-GAGA related protein (GRP) or empty vector were transfected into Jurkat cells with pREP4 episomal vector plasmids carrying the hsp90alpha promoter sequence from -1756 to +37 and control plasmids pMCAT. Total RNA was extracted. The relative promoter activity of hsp90alpha-CAT reporter gene was determined by competitive RT-PCR assay.

RESULTSGRP markly increased the relative promoter activity of hsp90alpha-CAT reporter gene during heat shock.

CONCLUSIONGRP may promote the expression of hsp90alpha gene by participating in chromatin remolding.

Amino Acid Motifs ; Animals ; Checkpoint Kinase 1 ; Cloning, Molecular ; DNA ; metabolism ; DNA, Complementary ; metabolism ; DNA-Binding Proteins ; metabolism ; Drosophila ; genetics ; Drosophila Proteins ; genetics ; Gene Expression Regulation ; HSP90 Heat-Shock Proteins ; biosynthesis ; genetics ; Heat-Shock Response ; genetics ; Homeodomain Proteins ; chemistry ; genetics ; metabolism ; Humans ; Repressor Proteins ; genetics ; Transcription Factors ; genetics ; Two-Hybrid System Techniques