Optimized combination of circulating biomarkers as predictors of prognosis in AECOPD patients complicated with Heart Failure.

Background: Systematic inflammation, nutritional status, and cardiovascular function have been associated with the outcomes of acute exacerbation of chronic obstructive pulmonary disease (AECOPD) patients with heart failure (HF). However, the value of their relevant biomarkers in predicting mortality has not been well defined yet. We aimed to investigate the prognostic value of circulating biomarkers including C-reaction protein (CRP)/albumin (ALB), neutrophil-lymphocyte ratio (NLR), platelet-lymphocyte ratio (PLR), and N-terminal pro-brain natriuretic peptide (NT-proBNP) for AECOPD patients with HF. Methods: A retrospective study was carried out in the Second Clinical College of Jinan University from January 1, 2013 to January 31, 2019. A total of 146 cases of AECOPD complicated with HF were enrolled and classified into survivor group (n=94) and non-survivor group (n=52). The baseline characteristics, CRP/ALB ratio, NLR, PLR, serum levels of NT-proBNP, and other indicators were collected. The predictors for prognosis were analyzed by multivariate logistic regression, and the ability to predict 28-day mortality was evaluated by receiver operating characteristics curve (ROC) and the area under the curve (AUC). Results: The patients in non-survivors had significantly higher levels of CRP, CRP/ALB, NLR, PCT and NT-proBNP, but lower ALB levels compared to the survivors [111.7 (56.9, 186.5) VS. 43.8 (10.3, 96.1) mg/L, 4.6 (2.0, 8.0) VS. 1.4 (0.3, 3.4), 22.2 (11.1, 40.1) VS. 12.0 (6.2, 24.8), 2.6 (0.2, 10.3) VS. 0.08 (0.1, 0.5) ng/ml, 17912.5 (9344.0, 34344.5) VS. 9809.0 (4415.9, 16387.2) ng/ml, 25.8 (23.2, 30.5) VS. 30.7 (27.9, 34.1) g/L; P < 0.001, <0.001, 0.001, <0.001, <0.001, and < 0.001, respectively]. No significant difference in PLR was found between the two groups (P=0.413). The logistic analysis revealed that CRP/ALB (OR=1.303, 95%CI: 1.145-1.483, P<0.001), NT-proBNP (OR=1.041, 95%CI: 1.010-1.073, P=0.009) and NLR (OR=1.010, 95%CI: 0.999-1.022, P<0.001) are independent risk factors for predicting the 28-day mortality. The AUC of the ROC curves were 0.768, 0.767, 0.757, 0.723, 0.716, and 0.668 for CRP/ALB, PCT, CRP, NT-proBNP, ALB, and NLR, respectively. The combination of CRP/ALB, NLR and NT-proBNP as biomarkers was shown to have better accuracy for predicting prognosis (AUC=0.830, 95%CI: 0.761-0.899, P<0.001), with a higher specificity of 80.8% and specificity of 77.7% as compared with each single biomarkers. Conclusions: High levels of NLR, CRP/ALB and NT-proBNP may be clinical usefully predictors for death in AECOPD patients with HF. Combination of NLR with CRP/ALB and NT-proBNP can provide a higher accuracy for predicting 28-day mortality in these patients.

Expression of herpesvirus entry mediator gene as a potential biomarker for disease severity in patients with persistent asthma.

Objectives: Herpes virus entry mediator (HVEM) is a costimulatory molecule, and has been proved to play an important role in airway inflammatory and remodeling processes of asthma. We aimed to investigate the expression of HVEM gene in patients with asthma as a means of assessing disease severity.Methods: This study was carried out on 59 subjects, 16 patients with mild persistent asthma, 11 patients with moderate persistent asthma, 13 patients with severe persistent asthma, and 19 age and gender matched healthy controls. The HVEM mRNA expressions of all subjects were determined by real time PCR. Correlations between HVEM mRNA expression and fractional exhaled nitric oxide (FeNO), pulmonary function test values, total blood white cell count and differential, total immunoglobulin E (IgE) level, and Asthma Control Test (ACT) score were analyzed, respectively. The discrimination abilities of HVEM mRNA between different groups were tested using receiver operating characteristics (ROC) curve analyses.Results: This study showed the expressions of HVEM mRNA were significantly higher in the patients with severe and moderate persistent asthma than in patients with mild persistent asthma and healthy subjects (2.97 ± 1.23 vs. 1.17 ± 0.42 vs. 0.62 ± 0.38 vs. 0.46 ± 0.18/NAPDH, p < 0.001), but there was no significant difference between patients with mild persistent asthma and health controls (0.62 ± 0.38 vs. 0.46 ± 0.18/NAPDH, p = 0.557). HVEM mRNA expression at cut off point [1.01/NAPDH, area under the ROC curve (AUC) = 0.99] is sufficient to discriminate severe patients from mild-to-moderate patients, and at cut off point (0.93/NAPDH, AUC = 0.91) for discrimination of moderate-to-severe patients from mild ones, while at cut off point (0.76/NAPDH, AUC = 0.75) for discrimination of asthmatic patients from controls. Furthermore, HVEM mRNA expression was positively correlated with FeNO level (r = 0.524, p = 0.015), and total lymphocyte count (r = 0.426, p = 0.017) in patients with persistent asthma.Conclusions: HVEM gene expressions can be used as a potential biomarker for evaluating the severity of patients with persistent asthma.

Molecular biology of BPIFB1 and its advances in disease.

Bactericidal/permeability-increasing (BPI)-fold-containing family B member 1 (BPIFB1), also known as long-palate lung and nasal epithelium clone 1 (LPLUNC1), belongs to the BPI-fold-containing family, is a newly discovered natural immune protection molecule, which, having the function of bactericidal and osmotic enhancement protein domain, can respond to the external physical and chemical stimuli. The gene of BPIFB1 is located at chromosome 20q11.21-20q11.22, and contains 16 exons and 15 introns, encoding 484 amino acids. The 5' terminal of the BPIFB1 protein has a signal peptide sequence composed of 19 amino acids. BPIFB1 is abnormally expressed in nasopharyngeal carcinoma (NPC), gastric cancer, and other cancer tissues, regulate chronic infections and inflammation, indicating that it may play an important role in the development of tumors. Meanwhile, BPIFB1 has well-recognized roles in sensing and responding to Gram-negative bacteria due to its structural similarity with BPI protein and lipopolysaccharide (LPS)-binding protein, both of which are innate immune molecules with recognized roles in sensing and responding to Gram-negative bacteria, so it can regulate cystic fibrosis (CF), chronic obstructive pulmonary disease (COPD), asthma, and other respiratory diseases. In this article, we will discuss the progress of BPIFB1 in a variety of diseases and fully understand its function.

The application of proteomics in the diagnosis and treatment of bronchial asthma.

Bronchial asthma is a common chronic inflammatory disease of the airways. Although its pathogenic mechanism remains unknown, it is influenced by both genetic and environmental factors. The emergence and application of proteomic technologies can help to facilitate analysis of the changes in transcription factors, inflammatory mediators, chemokines, cytokines, and cell apoptosis-and proliferation-related proteins in the pathological processes of asthma. Proteomic technologies can unearth prospects and theoretical bases for improved understanding of the biological mechanism of asthma and effective identification of diagnostic and therapeutic targets.

Advances in nanotechnology and asthma.

According to the World Health Organization, Asthma is the fastest-growing disease in the world alongside HIV/AIDS, and its socioeconomic burden exceeds the sum of HIV/AIDS and tuberculosis. Its high disability and mortality rates have become serious social and public health concerns. Asthma is a heterogeneous disease in which genetic polymorphisms interact with the environmental factors. While no specific treatment has been available for asthma due to its complex pathogenesis, the advances in nanotechnology have brought new hope for the early diagnosis, treatment, and prevention of asthma. Nanotechnology can achieve targeted delivery of drugs or genes, reduce toxic effects, and improve drug bioavailability. The nano-modifications of drugs and the development of new nano-drugs have become new research directions. Studies have demonstrated the safety and effectiveness of nanocarriers. However, many challenges still need to be overcome before nanotherapy can be applied in clinical practice. In this article we review the new research highlights in this area, with an attempt to explore the great potential and feasibility of nanotechnology in treating asthma.

Pulmonary lesions associated with sputum culture-positive actinomycetes: report of one case.

Pulmonary actinomycosis (PA) is a rare subacute or chronic infectious disease. As simple culture of Actinomyces in BAL, as with sputum, may represent colonization, the diagnosis of PA relies on pathological examination. The preferred treatment is long-term, high-dose penicillin. A 6-12-month-course of antibacterial treatment is the rule in extended PA, although the optimal duration of treatment has not been investigated through randomized trial. In this article, we report a case presented with slowly-progressing pulmonary cavitary lesions. Actinomyces odontolyticus was detected in sputum specimen harvested by tracheoscopy. The clinical diagnosis was PA, which gradually improved with prolonged treatment of penicillin and ornidazole. This is followed by a discussion of diagnosis and treatment, especially in terms of treatment.

Heat-Induced Epithelial Barrier Dysfunction Occurs via C-Src Kinase and P120ctn Expression Regulation in the Lungs.

BACKGROUND/AIMS: Thermal injury causes pulmonary edema and can lead to death. Intercellular junctions are composed of adhesive (p120ctn, E-cadherin, α-catenin and ß-catenin) and compact (occludin and ZO-1) junctions. Heat deteriorates intercellular junctions and increases cell gaps to ultimately induce pulmonary edema, but the underlying mechanism remains elusive. METHODS: Mouse lung epithelial (MLE-12) cells pre-treated with the c-Src inhibitor PP2, p120ctn catenin (p120ctn) small interfering RNA and p120ctn catenin (p120ctn) complementary DNA were subjected to heat treatment. Western blotting and real-time polymerase chain reaction assays were used to evaluate junction protein expression changes after heat treatment, and co-immunoprecipitation was used to test the binding state of junction proteins. In addition, hematoxylin and eosin staining and immunohistochemistry were used to evaluate changes in junction protein expression and lung injury in a Wistar rat model of thermal inhalation injury. RESULTS: Heat increased cell permeability; induced ZO-1, occludin, α-catenin and ß-catenin degradation; and decreased E-cadherin distribution in cell membranes. Heat also activated c-Src and decreased both p120ctn expression levels and occludin and ZO-1 association. c-Src inhibitor (PP2) treatment and p120ctn overexpression reversed these effects and attenuated lung injury in vivo. CONCLUSION: Heat induces junction protein degradation and dissociation to increase membrane permeability and cause lung edema via c-Src kinase activation and p120ctn expression downregulation.

Comparison of Fractional Exhaled Nitric Oxide in Elderly Patients with Asthma-chronic Obstructive Pulmonary Disease Overlap and Other Airway Inflammatory Diseases.

The exact role of fractional exhaled nitric oxide (FeNO) in older patients with chronic inflammatory diseases including asthma-chronic obstructive pulmonary disease (COPD) overlap (ACO) remains unclear. This study aimed to investigate the differences in FeNO levels of elderly patients with ACO, asthma, COPD, and chronic cough. We conducted a retrospective study analysing the data of stable outpatients from Pulmonary Department of the Second Clinical College, Jinan University. All participants (Age≥55 years) were divided into the ACO group (n=19), asthma group (n=16), COPD group (n=25), and chronic cough group (n=22). The clinical data such as peripheral eosinophil counts, serum high sensitivity C-reactive protein (hs-CRP), FeNO, and spirometry was collected, and the correlations between FeNO levels and systemic markers or spirometric indices were analyzed. Patients with ACO and asthma had significantly elevated FeNO levels (37.7±16.5, and 36.3±17.7 ppb) compared with COPD, and chronic cough patients (21.9±10.3, and 16.1±8.8 ppb). The FeNO levels were negatively associated with forced expiratory volume in 1 second (FEV1, p=0.003), FEV1% predicted (p=0.012), and FEV1/forced vital capacity (FVC, p=0.002) in all groups. However, there were no significant correlation between FeNO levels and FVC, peripheral eosinophil counts, or serum hs-CRP (p>0.05). Elderly patients with ACO have higher levels of FeNO, when compared with patients with COPD or chronic cough. These findings suggest that FeNO measurement may provide an important implication for the etiological diagnosis of ACO in the elderly patients.

Suppression of OsMADS7 in rice endosperm stabilizes amylose content under high temperature stress.

High temperature significantly alters the amylose content of rice, resulting in mature grains with poor eating quality. However, only few genes and/or quantitative trait loci involved in this process have been isolated and the molecular mechanisms of this effect remain unclear. Here, we describe a floral organ identity gene, OsMADS7, involved in stabilizing rice amylose content at high temperature. OsMADS7 is greatly induced by high temperature at the early filling stage. Constitutive suppression of OsMADS7 stabilizes amylose content under high temperature stress but results in low spikelet fertility. However, rice plants with both stable amylose content at high temperature and normal spikelet fertility can be obtained by specifically suppressing OsMADS7 in endosperm. GBSSI is the major enzyme responsible for amylose biosynthesis. A low filling rate and high expression of GBSSI were detected in OsMADS7 RNAi plants at high temperature, which may be correlated with stabilized amylose content in these transgenic seeds under high temperature. Thus, specific suppression of OsMADS7 in endosperm could improve the stability of rice amylose content at high temperature, and such transgenic materials may be a valuable genetic resource for breeding rice with elite thermal resilience.

Perovskite-type calcium titanate nanoparticles as novel matrix for designing sensitive electrochemical biosensing.

In this work, novel perovskite-type calcium titanate nanoparticles (CaTiO3NPs) were for the first time exploited for the immobilization of proteins and the development of electrochemical biosensor. The CaTiO3NPs were synthesized with a simple and cost-effective route at low temperature, and characterized by scanning electron microscopy, X-ray photoelectron spectroscopic spectrum, electrochemical impedance spectrum, UV-visible spectroscopy, Fourier transform infrared spectrum, and cyclic voltammetry, respectively. The results indicated that CaTiO3NPs exhibited large surface area, and greatly promoted the direct electron transfer between enzyme molecules and electrode surface. The immobilized enzymes on this matrix retained its native bioactivity and exhibited a surface controlled, quasi-reversible two-proton and two-electron transfer reaction with an electron transfer rate of 3.35s-1. Using glucose oxidase as model, the prepared glucose biosensor showed a high sensitivity of 14.10±0.5mAM-1 cm-2, a wide linear range of 7.0×10-6 to 1.49×10-3M, and a low detection limit of 2.3×10-6M at signal-to-noise of 3. Moreover, the biosensor also possessed good reproducibility, excellent selectivity and acceptable storage life. This research provided a new-type and promising perovskite nanomaterials for the development of efficient biosensors.

MicroRNA-181a, -146a and -146b in spleen CD4+ T lymphocytes play proinflammatory roles in a murine model of asthma.

CD4+ T lymphocytes can be primarily polarized to differentiate into Th2 cells, and are heavily involved in the Th2 inflammation of asthma. Little is known about the correlation between microRNAs and Th2 inflammation in asthma, therefore we explore the roles of five microRNAs (microRNA-181a, -155, -150, -146a and -146b) in Th2 inflammation of asthma by tracking their expression levels in splenic CD4+ T lymphocytes under different conditions. Using quantitative real-time polymerase chain reaction (qPCR), the dynamic changes of these microRNAs in murine models of acute asthma (i.e. the OVA group) were analyzed, in comparison to a control group. The effects of dexamethasone on the miRNA expression levels were also investigated. The results showed that the expression levels of microRNA-181a, -150, -146a and -146b were higher in the OVA group compared to the control group in the beginning of the disease, and after 5days dropped to control group levels because there was no new airway challenge. Moreover, the miRNA-146a expression was down-regulated by treatment with dexamethasone. MicroRNA-181a had a positive linear correlation with the numbers of inflammatory cells (i.e. the numbers of total cells or of the eosinophils in the BALF) by Spearman correlation analysis, so did miRNA-146a and miRNA-146b. These observations suggest that microRNA-181a, -146a and -146b are proinflammatory factors in asthma, and that down-regulation of miRNA-146a may partially account for the anti-inflammatory effect of dexamethasone.

[Proliferation of T lymphocyte subsets in asthma patients in clinical remission and molecular mechanism thereof].

OBJECTIVE: To explore the rule of proliferation of T lymphocyte subsets in patients with clinical asthma remission and the molecular mechanism. METHODS: Peripheral blood samples were collected from 15 asthmatic patients, 15 asthmatic patients in clinical remission, and 15 healthy control subjects, all sex-, and age-matched. CD(4)(+) T and CD(8)(+) T lymphocytes were isolated. Flow cytometry was used to examine the cell cycles of CD(4)(+) T and CD(8)(+) T lymphocytes. Fluorescence immunohistochemistry was used to detect the expression levels of cell cycle regulatory proteins (CCRPs), including cyclin D, cyclin E, and P27(kip1), PI3K, and STAT6. RESULTS: (1) The percentage of G(0)/G(1) phase of the CD(4)(+) T lymphocytes of the asthmatic patients was 82.00%, significantly lower than those of the asthmatic patients in clinical remission and healthy controls (92.50% and 99.00%, Z = 12.35, P < 0.01). The percentage of S phase of the CD(4)(+) T lymphocytes of the asthmatic patients was 18.00%, significantly higher than those of the asthmatic patients in clinical remission and healthy controls (6.10% and 0.20% respectively, Z = 8.05, P < 0.05). The percentage of G(2)/M phase of the CD(4)(+) T lymphocytes of the asthmatic patients was 2.80%, significantly higher than those of the asthmatic patients in clinical remission and healthy controls (0.40% and 0 respectively, Z = 9.16, P < 0.05). The S + G(2)/M phase of the CD(4)(+) T lymphocytes of the asthmatic patients was 18.00%, significantly higher than those of the asthmatic patients in clinical remission and healthy controls (7.50% and 0.20% respectively, Z = 12.80, P < 0.05). The distribution of G(0)/G(1) phase of CD(8)(+) T lymphocyte of the asthmatic patients was 44.60%, significantly lower than those of the asthmatic patients in clinical remission and healthy controls (95.90% and 100.00% respectively, Z = 21.60, P < 0.01). The distribution of S phase of CD(8)(+) T lymphocytes of the asthmatic patients was 51.70%, significantly lower than those of the asthmatic patients in clinical remission and healthy controls (0.80% and 0 respectively, Z = 25.22, P < 0.01). The distribution of S + G(2)/M phase of the CD(8)(+) T lymphocytes of the asthmatic patients was 55.40%, significantly higher than those of the asthmatic patients in clinical remission and healthy controls (4.10% and 0 respectively, Z = 21.52, P < 0.01). (2) The expression level of P27(kipl) of the CD(4)(+) T lymphocytes of the asthmatic patients was 13.20%, significant lower than those of the asthmatic patients in clinical remission and healthy controls (38.80% and 47.20% respectively, Z = 10.63, P < 0.01). The expression level of cyclin D of the CD(4)(+) T lymphocyte of the asthmatic patients was 35.00%, significant higher than those of the asthmatic patients in clinical remission and healthy controls (28.20% and 13.10% respectively, Z = 10.66, P < 0.01). The expression level of cyclin E of the CD(4)(+) T lymphocytes of the asthmatic patients was 7.90%, significant higher than those of the asthmatic patients in clinical remission and healthy controls (6.30% and 3.70% respectively, Z = 6.64%, P < 0.05). The expression level of P27(kipl) of the CD(8)(+) T lymphocyte of the asthmatic patients was 4.50%, significant lower than those of the asthmatic patients in clinical remission and healthy controls (33.80% and 46.30% respectively, Z = 9.30, P < 0.05). The expression level of cyclin D of the CD(8)(+) T lymphocyte of the asthmatic patients was 24.20%, not significant different from those of the asthmatic patients in clinical remission and healthy controls (26.10% and 32.20% respectively, Z = 0.09, P > 0.05). The expression level of cyclin E of the CD(8)(+) T lymphocyte of the asthmatic patients was 9.30%, significant higher than those of the asthmatic patients in clinical remission and healthy controls (5.60% and 3.50% respectively, Z = 4.91, P > 0.05). (3) The expression level of PI(3)K-110alpha of the CD(4)(+) T lymphocyte of the asthmatic patients was 7.60%, significant higher than those of the asthmatic patients in clinical remission and healthy controls (6.40% and 3.30% respectively, Z = 9.04, P < 0.05). The expression level of STST()6 of the CD(4)(+) T lymphocyte of the asthmatic patients was 8.20%, significant higher than those of the asthmatic patients in clinical remission and healthy controls (2.70% and 1.90% respectively, Z = 18.08, P > 0.01). The expression levels of PI(3)K-110alpha and STST(6) of the CD(8)(+) T lymphocytes of the asthmatic patients were not significant different from those of the asthmatic patients in clinical remission and healthy controls (Z = 4.91 and 5.70, both P > 0.05). CONCLUSION: There is excessive proliferation of CD(4)(+) T lymphocytes in the patients with clinical asthma remission, which may be related to the abnormal expression of CCRP (cyclin D, cyclin E, and P27(kip1)), PI(3)K, and STAT(6).

[The effect of phosphoinositide-3-kinase and signal transducer and activator of transcription-6 on the proliferation of T lymphocytes in bronchial asthma].

OBJECTIVE: To investigate the effect of phosphoinositide-3-kinase (PI(3)K) and signal transducer and activator of transcription-6 (STAT(6)) on the proliferation of CD(4)(+) and CD(8)(+) T lymphocytes in bronchial asthma. METHODS: CD(4)(+) and CD(8)(+) T lymphocytes of 15 asthmatic patients or 15 healthy control subjects were cultured in vitro from August of 2004 to February of 2005. The T lymphocytes of asthmatic patients were divided into four groups and stimulated with or without 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002) and IFN-gamma. The four groups included a control group (group A), the LY294002 group (group B), IFN-gamma group (group C) and LY294002 + IFN-gamma group (group D). The cell cycle phases and the expression of cell cycle proteins (P27kip1, Cyclin D, Cyclin E), PI(3)K and STAT(6) were analyzed by flow cytometry. RESULTS: (1) The percentage of G(0)/G(1) phase, the expression rate of P27kip1 in CD(4)(+) and CD(8)(+) T lymphocytes from asthmatic patients were 82.0%, 44.6%, 13.2%, 4.5%; and those from the control group were 99.0%, 100.0%, 47.2%, 46.3%, respectively; the difference was significant between the two groups (Z value were 3.54, 4.23, 3.09 and 2.51, all P < 0.05). The percentage of S phase, the expression rate of Cyclin E, PI(3)K and STAT(6) in CD(4)(+) and CD(8)(+) T lymphocytes from the asthmatic patients were 18.0%, 51.7%, 7.9%, 9.3%, 7.6%, 8.7%, 8.2%, 6.3%; and those from the control group were 0.2%, 0.0%, 3.7%, 3.5%, 3.3%, 3.4%, 1.9%, 2.4%, respectively; there were significant differences between the two groups (Z value were 2.88, 4.61, 1.95, 2.06, 2.51, 2.32, 4.38 and 2.22, all P < 0.05). (2) The percentage of G(0)/G(1) phase, S phase, the expression rate of Cyclin D, Cyclin E in CD(4)(+) T lymphocytes of B group were 95.6%, 1.9%, 13.3% and 3.1%; and those of A group were 82.0%, 18.0%, 35.0%, 7.9%; there were significant differences between the two groups (Z value were 2.04, 2.23, 2.78 and 1.99, all P < 0.05). The percentage of S phase, the expression rate of Cyclin E in CD(8)(+) T lymphocytes of B group were 1.0% and 4.1%; and those of A group were 51.7% and 9.3%; there were significant differences between the two groups (Z value were 3.06 and 2.56, all P < 0.05). The percentage of G(0)/G(1) phase, S phase, the expression rate of P27kip1 in CD(4)(+) T lymphocyte of C group were 94.0%, 1.5%, 46.1%; and those of A group were 82.0%, 18.0%, 13.2%; there were significant differences between the two groups (Z value were 2.17, 2.54 and 2.81, all P < 0.05). The percentage of S phase, the expression rate of P27kip1 in CD(8)(+) T lymphocyte of C group were 10.8% and 23.1%; and those of A group were 51.7% and 4.5%; there were significant differences between the two groups (Z value were 2.67 and 2.05, all P < 0.05). The percentage of G(0)/G(1) phase, S phase, the expression rate of P27kip1, Cyclin D in CD(4)(+) T lymphocytes of D group were 97.0%, 0.0%, 40.4%, 21.5%; and those of A group were 82.0%, 18.0%, 13.2%, 35.0%; there were significant differences between the two groups (Z value were 2.73, 2.79, 2.56 and 2.10, all P < 0.05). The percentage of S phase, the expression rate of P27kip1 in CD(8)(+) T lymphocytes of D group were 92.1% and 1.7%; and those of A group were 44.6% and 51.7%; there were significant differences between the two groups (Z were 2.22 and 3.12, all P < 0.05). CONCLUSION: PI(3)K and STAT(6) may be new therapeutic targets for the treatment of asthma. Further studies are necessary to explore the relationship between PI(3)K and JAK1-STAT(6) signal pathway during the enhancement of CD(4)(+) T or CD(8)(+) T lymphocyte proliferation in bronchial asthma.

[Application of immunomagnetic screening strategy for separation of CD4+ and CD8+ T cell subpopulations of peripheral blood].

To evaluate the separation of T lymphocyte subsets by immunomagnetic beads and to find optimization of strategy for specific binding of antibody-coated beads to cells, two strategies to isolate enriched T lymphocyte subpopulation CD4+ T cells and CD8+ T cells from small volumes (< 5 ml) of peripheral blood by using immunomagnetic beads or complement cytotoxicity method were compared. The purity and activity of CD4+ T cells and CD8+ T cells were measured by using flow cytometry, trypan-blue dye exclusion test, etc. The results showed that the yields of CD4+ T lymphocytes and CD8+ T lymphocytes by using immunomagnetic beads were (94.2 +/- 1.4)% and (93.8 +/- 3.0)% respectively, higher than those of control group and the group of using completement cytotoxicity method (P < 0.05). At the same time, the yields of CD4+ T lymphocytes and CD8+ T lymphocytes by using complement cytotoxicity method were (76.0 +/- 2.8)% and (77.0 +/- 3.0)% respectively, higher than those of unenriched group (P < 0.05). The trypan-blue dye exclusion test confirmed that there were no influences on activity of CD4+ T cells and CD8+ T cells when immunomagnetic beads were used for separation of these cells from peripheral blood. It is concluded that the immunomagnetic bead method has a higher efficiency for separation of CD4+ T cells and CD8+ T cells from peripheral blood than complement cytotoxic method, especially for small sample. This method has no influence on activity and proliferation of T lymphocyte subpopulations, and would be expected to establish conditions for research of biological characteristics of CD4+ T cells and CD8+ T cells in future.