RNA 5-Methylcytosine Facilitates the Maternal-to-Zygotic Transition by Preventing Maternal mRNA Decay.

The maternal-to-zygotic transition (MZT) is a conserved and fundamental process during which the maternal environment is converted to an environment of embryonic-driven development through dramatic reprogramming. However, how maternally supplied transcripts are dynamically regulated during MZT remains largely unknown. Herein, through genome-wide profiling of RNA 5-methylcytosine (m5C) modification in zebrafish early embryos, we found that m5C-modified maternal mRNAs display higher stability than non-m5C-modified mRNAs during MZT. We discovered that Y-box binding protein 1 (Ybx1) preferentially recognizes m5C-modified mRNAs through π-π interactions with a key residue, Trp45, in Ybx1's cold shock domain (CSD), which plays essential roles in maternal mRNA stability and early embryogenesis of zebrafish. Together with the mRNA stabilizer Pabpc1a, Ybx1 promotes the stability of its target mRNAs in an m5C-dependent manner. Our study demonstrates an unexpected mechanism of RNA m5C-regulated maternal mRNA stabilization during zebrafish MZT, highlighting the critical role of m5C mRNA modification in early development.

The ability of volumetric capnography to distinguish between chronic obstructive pulmonary disease patients and normal subjects.

PURPOSE: The aim of our study was to evaluate volumetric capnography (VCap) in the differentiation between chronic obstructive pulmonary disease (COPD) patients and normal subjects. PATIENTS AND METHODS: Thirty-nine healthy male volunteers and 60 male COPD patients were enrolled. Regression equations between VCap indices and age, weight, height, and tidal volume in healthy volunteers were established by stepwise regression analysis. Predicted normal values of VCap indices in COPD patients were calculated. A paired t test was used to compare the difference between observed and predicted values for VCap indices in COPD patients. Receiver operating characteristic (ROC) curve analysis was used to evaluate the power of each VCap index alone in differentiating COPD patients and normal subjects. The power of the combination of VCap indices was assessed by discriminant analysis. RESULTS: All regression equations were significant (P < 0.01) as were the differences between the observed and predicted normal VCap indices in COPD patients (P < 0.001). ROC curve analysis showed that the volume between 25 and 50% of F CO2et (Vm25-50), slope of Phase II (dC2/dV), and slope of Phase III (dC3/dV) were valuable predictors. Nearly all (90.9%) subjects were correctly classified by discriminant analysis. CONCLUSION: Vm25-50, dC2/dV, or dC3/dV alone are valuable for differentiating COPD patients and normal subjects, but more powerful are the combinations of Vm25-50, dC2/dV, and dC3/dV, the ratio of dC2/dV to dC3/dV (SR23), dead space according to the Bohr method (VDB), and dead space according to the Wolff and Brunner methods (PIE).

Detection of the airway obstruction stage in asthma using impulse oscillometry system.

OBJECTIVE: Although spirometry is the most common method for evaluating the airway obstruction stage in asthma patients, it is difficult to perform in some patients. The aim of this study was to evaluate whether impulse oscillometry, an easy-to-perform technique, can detect asthmatic airway obstruction stage. METHODS: A total of 80 subjects, including healthy volunteers and patients with asthma, were enrolled in this study. The asthma patients were classified into three groups according to American Thoracic Society (ATS)/European Respiratory Society (ERS)-2005: the mild group (forced expiratory volume in 1 second (FEV(1)) ≥ 70% predicted (Pred), n = 20), the moderate group (50% Pred ≤ FEV(1) <70% Pred, n = 20), and the severe group (FEV(1) < 50% Pred, n = 20). Spirometry and impulse oscillometry (IOS) parameters were obtained from every subject. Correlation analysis was used to compare spirometry measurements and IOS parameters. One-way analysis of variance (ANOVA) was performed to compare IOS parameters among different groups. The potential of using all individual IOS parameters to detect the different stages of asthmatic airway obstruction was evaluated by the receiver operating characteristic (ROC) curve analysis. RESULTS: The correlation analysis showed that IOS parameters, such as respiratory resistance at 5 Hz (R(5)), respiratory resistance at 10 Hz (R(10)), respiratory resistance at 20 Hz (R(20)), difference in resistance between 5 Hz and 20 Hz (R(5-20)), impedance at 5 Hz (Z(5)), resonant frequency (Fres), and area of reactance (AX) were negatively correlated with FEV(1) and peak expiratory flow (PEF), while reactance at 5 Hz (X(5)) was positively correlated with FEV(1) and PEF. The increase in R(5), R(10), R(20), Z(5), R(5-20), Fres, (-X(5)), and AX parameters corresponded significantly with an increase in the airway obstruction stage, as determined by one-way ANOVA . ROC curve analysis revealed that all the IOS parameters studied, except for Fres, were capable of classifying asthmatic airway obstruction. R(5), R(5-20), and Z(5) were the most accurate parameters. CONCLUSION: IOS provides an accurate, reliable, and patient-friendly technique for classifying asthmatic airway obstruction.

[Association of oxygen uptake efficiency in exercise test with exercise capacity in patients with stable chronic obstructive pulmonary disease].

OBJECTIVE: To explore the characteristics of oxygen uptake efficiency (OUE) in patients with chronic obstructive pulmonary disease (COPD), and to analyze the relationship between OUE and the exercise capacity. METHODS: Pulmonary function test and cardiopulmonary exercise test were performed in 59 patients with stable COPD (grade I, n = 15; grade II, n = 16; grade III, n = 19; grade IV, n = 9) and 29 healthy volunteers of the same age. Their successive breathing respiratory exchange parameters were collected and analyzed. t test and χ(2) test were used for 2 sample comparison, while multiple comparisons among groups were performed by using single factor analysis of variance. Correlation analysis was done by Pearson correlation test. RESULTS: Compared with the normal control group [(2.2 ± 0.4) L·min(-1)·lg(L·min(-1))(-1); (35 ± 4) ml/L], the OUES and OUEP of the COPD patient group [(1.9 ± 0.3) L·min(-1)·lg(L·min(-1))(-1); (31 ± 5) ml/L]were significantly lower (t = 4.57, 3.39, all P < 0.01) . The OUE of the grade I patients showed no significant difference compared with the normal control group (t = 0.36-1.49, all P > 0.05), while the OUES of the grade II-IV patients [(2.05 ± 0.26), (1.76 ± 0.28) and (1.63 ± 0.19) L·min(-1)·lg(L·min(-1))(-1)] decreased significantly compared to the normal control group [(2.23 ± 0.39) L·min(-1)·lg(L·min(-1))(-1); t = 2.42-5.26, all P < 0.05]. The OUEP and the OUE at the anaerobic threshold of the grade II-III patients [(31 ± 4) and (31 ± 5), (29 ± 5) and (29 ± 5) ml/L] decreased significantly compared to the normal control group [(35 ± 4) and (34 ± 4) ml/L, t = 2.18-4.83, all P < 0.05]. The OUES, OUEP and the OUE at the anaerobic threshold in COPD patients were correlated (r = 0.500-0.625, all P < 0.01) positively with the exercise tolerance (peak VO2% pred). CONCLUSIONS: The oxygen uptake efficiency of patients with COPD is significantly reduced compared to that of the normal subjects, and is correlated positively with the exercise capacity.

[Effects of whole lung lavage on pulmonary function and exercise capacity in patients with pneumoconiosis].

OBJECTIVE: To investigate the effects of whole lung lavage (WLL) on the pulmonary function and exercise capacity in patients with pneumoconiosis. METHODS: Forty-one patients with pneumoconiosis who quit dust-exposed work not more than 6 months before underwent WLL. Clinical symptom assessment, pulmonary function test, and cardiopulmonary exercise test were performed before and one week after WLL, and the results were compared. RESULTS: The patients with pneumoconiosis showed no significant changes in clinical symptoms after WLL. At one week after WLL, the patients with pneumoconiosis showed nonsignificant increases in forced vital capacity, forced expiratory volume in one second (FEV1.0), and percent predicted FEV1 (P > 0.05); peak oxygen uptake (peak VO2) increased from 2140.6 ± 353.2 ml/min before WLL to 2374.6 ± 362.4 ml/min after WLL, percent predicted peak VO2 increased from 82.2 ± 13.7% before WLL to 91.0 ± 14.0% after WLL, peak VO2/kg increased from 30.6 ± 3.5 ml/min×kg before WLL to 34.2 ± 3.7 ml/min×kg after WLL, and ventilatory equivalent for carbon dioxide decreased from 30.6 ± 3.1 before WLL to 26.1 ± 2.7 after WLL (P < 0.05). CONCLUSION: WLL can remarkably improve the oxygen uptake and ventilatory efficiency in patients with pneumoconiosis during exercise, so it can improve the exercise capacity of these patients.

Nsun2 coupling with RoRγt shapes the fate of Th17 cells and promotes colitis.

T helper 17 (Th17) cells are a subset of CD4+ T helper cells involved in the inflammatory response in autoimmunity. Th17 cells secrete Th17 specific cytokines, such as IL-17A and IL17-F, which are governed by the master transcription factor RoRγt. However, the epigenetic mechanism regulating Th17 cell function is still not fully understood. Here, we reveal that deletion of RNA 5-methylcytosine (m5C) methyltransferase Nsun2 in mouse CD4+ T cells specifically inhibits Th17 cell differentiation and alleviates Th17 cell-induced colitis pathogenesis. Mechanistically, RoRγt can recruit Nsun2 to chromatin regions of their targets, including Il17a and Il17f, leading to the transcription-coupled m5C formation and consequently enhanced mRNA stability. Our study demonstrates a m5C mediated cell intrinsic function in Th17 cells and suggests Nsun2 as a potential therapeutic target for autoimmune disease.

Clinical efficacy of tetrandrine in artificial stone-associated silicosis: A retrospective cohort study.

Background: Outbreaks of silicosis have occurred among workers in the artificial stone (AS) industry, and there is currently no effective antifibrosis treatment for silicosis. Design: A retrospective cohort study. Methods: We retrospectively analyzed the clinical data of 89 artificial stone-associated silicosis patients treated in Shanghai Pulmonary Hospital (China). Patients who agreed to be administered tetrandrine entered the observation group and those who disagreed entered the control group. Changes in chest HRCT, pulmonary function, and clinical symptoms of patients in two groups were compared pre- and post-treatment. Results: After treatment for 3-12 months, 56.5%-65.4% of patients in the observation group showed improvements in HRCT imaging, while there was no improvement in the control group (p < 0.05). Disease progression occurred in 0%-17.4% of patients in the observation group after 3-12 months of treatment compared with 44.4%-92.0% of patients in the control group (p < 0.05). After 3 months of treatment, the forced vital capacity (FVC), forced expiratory volume in 1 s (FEV1), and diffusing capacity of the lung for carbon monoxide (DLco) in the observation group increased by 136.7 ± 189.2 mL (p < 0.05), 124.2 ± 169.9 mL (p < 0.05), and 1.4 ± 2.3 mL/min/mmHg (p > 0.05), respectively, while those in the control group decreased (145.8 ± 356.5; 107.5 ± 272.1; 1.9 ± 3.8). After 6 months of treatment, FVC, FEV1, and DLco in the observation group increased by 207.8 ± 372.2 mL (p > 0.05), 107.8 ± 295.2 mL (p > 0.05) and 0.7 ± 6.0 mL/min/mmHg (p > 0.05), respectively, while those of the control group decreased (383.3 ± 536.7; 215.6 ± 228.9; 1.4 ± 1.7). The incidences of clinical symptoms such as cough, expectoration, dyspnea, chest tightness, and chest pain in the observation group were decreased-after treatment (all p < 0.05), while the incidences of these symptoms increased in the control group, although the change was not statistically significant (all p > 0.05). Conclusion: Tetrandrine can control and delay the progression of AS-associated silicosis fibrosis, with improved chest HRCT imaging and pulmonary function.

Dynamic transcriptomic m5 C and its regulatory role in RNA processing.

RNA 5-methylcytosine (m5 C) is a prevalent RNA modification in multiple RNA species, including messenger RNAs (mRNAs), transfer RNAs (tRNAs), ribosomal RNAs (rRNAs), and noncoding RNAs (ncRNAs), and broadly distributed from archaea, prokaryotes to eukaryotes. The multiple detecting techniques of m5 C have been developed, such as m5 C-RIP-seq, miCLIP-seq, AZA-IP-seq, RNA-BisSeq, TAWO-seq, and Nanopore sequencing. These high-throughput techniques, combined with corresponding analysis pipeline, provide a precise m5 C landscape contributing to the deciphering of its biological functions. The m5 C modification is distributed along with mRNA and enriched around 5'UTR and 3'UTR, and conserved in tRNAs and rRNAs. It is dynamically regulated by its related enzymes, including methyltransferases (NSUN, DNMT, and TRDMT family members), demethylases (TET families and ALKBH1), and binding proteins (ALYREF and YBX1). So far, accumulative studies have revealed that m5 C participates in a variety of RNA metabolism, including mRNA export, RNA stability, and translation. Depletion of m5 C modification in the organism could cause dysfunction of mitochondria, drawback of stress response, frustration of gametogenesis and embryogenesis, abnormality of neuro and brain development, and has been implicated in cell migration and tumorigenesis. In this review, we provide a comprehensive summary of dynamic regulatory elements of RNA m5 C, including methyltransferases (writers), demethylases (erasers), and binding proteins (readers). We also summarized the related detecting technologies and biological functions of the RNA 5-methylcytosine, and provided future perspectives in m5 C research. This article is categorized under: RNA Processing > RNA Editing and Modification.

Value of heart rate recovery in female patients with pulmonary arterial hypertension due to systemic lupus erythematosus.

OBJECTIVE: This study intended to explore the relation between heart rate recovery at 1 minutes (HRR1) during the recovery phase of cardiopulmonary exercise test (CPET) and exercise capacity in female systemic lupus erythematosus associated pulmonary arterial hypertension (SLE-PAH) patients. METHODS: Twenty-one female SLE-PAH patients underwent right heart catheterization (RHC), pulmonary function test (PFT) and CPET. Forty-two healthy subjects matched with SLE-PAH patients in age, sex and BMI were recruited as a control group. The correlations between HRR1 with clinical and CPET parameters were performed. RESULTS: Peak HR, ΔHR, HRR1, Peak HR-warm HR1min , Peak HR-warm HR2min and CR were significantly lower in SLE-PAH than in controls (P < .01). Increased incidence of CRI was seen in SLE-PAH. Except for the Peak PET O2 , which was higher in controls, all other CPET parameters were lower in SLE-PAH. SLE-PAH patients with HRR1 ≥ 16 had longer 6MWD, lower NT-proBNP, better percent of predicted gas transfer index or diffusing capacity for carbon monoxide (DLco% pred) as well as better CO and CI. Peak HR, ΔHR, HRR1, Peak HR-warm HR1min , Peak HR-warm HR2min , CR, Peak Load, Peak VO2 , Peak PET CO2 , OUEP and OUES were lower and duration of exercise was shorter in patients with HRR1 < 16. HRR1 had positive correlation with 6MWD, DLco% pred, CO, CI and some key CPET parameters. CONCLUSIONS: HRR1 is an easily obtained auxiliary parameter in SLE-PAH patients to reflect an altered autonomic tone. SLE-PAH patients with HRR1 < 16 have more severe hemodynamics, worse clinical findings and marked oxygen uptake inefficiency than those with HRR1 ≥ 16.

[The significance of volumetric capnography in assessment of asthmatic acute exacerbation staging].

OBJECTIVE: To investigate the diagnostic value of volumetric capnography in the assessment of asthmatic exacerbation. METHODS: Sixty-four patients with asthma exacerbation and 20 normal controls performed spirometry and volumetric capnography recording. The patients with asthma were divided into three sub-groups according to FEV1% pred (A: > 80%, B:40% - 80%, C: < 40%). RESULTS: FEV1% pred, FEV1/FVC, PEF% pred and MMEF% pred were (98 +/- 9)%, (80 +/- 6)%, (91 +/-15)% and (73 +/- 7)% respectively in the control group, but were (52 +/- 20)%, (50 +/- 10)%, (49 +/-16)% and (28 +/- 16)% respectively in the asthma group, the difference being significant (t = 6.93 - 13.29, all P < 0.01). Compared with the control group, dC2/DV [(19 +/- 6)%/L vs (31 +/- 8)%/L, t = 5. 09, P < 0.01] showed a decrease in the asthma group, dC3/DV [(2.9 +/- 1.2)%/L vs (1.0 +/- 0.4)%/L, t = -6. 14, P < 0.01] and SR23 [(16.8 +/- 10.6)% vs (3.3 +/-1.5)%, t = -6.54, P < 0.01] showed an increase in the asthma group compared to the control group, the difference being significant. Compared with that of the control group, dC2/DV [B:(17 +/-5)%/L, C: (13 +/-4)%/L] showed a decrease (t = -11.82, -16.75, all P < 0.01) and dC3/DV [B:(3.2 +/- 0.8)%/L, C:(4.1 +/-1.2)%/L] and SR23 [B:(17.2 +/- 3.5)%, C:(28.3 +/- 6.9)%] showed an increase (t = 2. 16-26.08, all P < 0.01) in asthma sub-groups B and C. For dC3/DV and SR23, the difference was significant between asthma sub-groups (t = 0.91 -22.18, all P < 0.05). In Pearson correlation analysis, dC2/DV (r = 0.69, 0.54, 0.59, 0.54, all P < 0.01) and dC3/DV (r = -0.62, -0.45, -0.69, -0.58, all P < 0.01) and SR23 (r = -0.75, -0.52, -0.74, -0.62, all P < 0.01) correlated with FEV1% pred, FEV1/FVC, PEF% pred and MMEF% pred. CONCLUSION: Volumetric capnography is a quantitative method for evaluating the severity of airflow obstruction and it can be performed easily during tidal breathing.

Adaptive Fibrogenic Reprogramming of Osteosarcoma Stem Cells Promotes Metastatic Growth.

It is well established that fibrotic remodeling of the tumor microenvironment favors tumorigenesis, but whether fibrosis underlies malignant progression in other ways is unclear. Here, we report that adaptive myofibroblastic reprogramming of osteosarcoma stem cells (OSCs) results in a critical advantage when establishing lung macro-metastases and spheroid growth but does not affect the growth of primary lesions or monolayer cultures. FGFR2 signaling in OSCs initiates fibrosis, whereas the resultant fibronectin (FN) auto-deposition sustains fibrogenic reprogramming and OSC growth, resembling the process employed by non-malignant myofibroblasts to cause tissue fibrosis. Furthermore, we provide evidence that nintedanib targets the pan FGFR-FN axis to disrupt lung metastasis without affecting the bone lesion growth of OSCs. Thus, myofibroblastic reprogramming of human OSCs in the lungs might represent a druggable trait for treating a deadly metastatic complication.

Mid-term effects of lung volume reduction surgery on pulmonary function in patients with chronic obstructive pulmonary disease.

BACKGROUND: Now lung volume reduction surgery (LVRS) has become one of the most effective methods for the management of some cases of severe chronic obstructive pulmonary disease (COPD). We evaluated the mid-term effects of LVRS on pulmonary function in patients with severe COPD. METHODS: Ten male patients with severe COPD aged 38 - 70 years underwent LVRS and their pulmonary function was assessed before, 3 months and 3 years after surgery. The spirometric and gas exchange parameters included residual volume, total lung capacity, inspiratory capacity, forced vital capacity, forced expiratory volume in one second, diffusion capacity for CO, and arterial blood gas. A 6-minute walk distance (6MWD) test was performed. RESULTS: As to preoperative assessment, most spirometric parameters and 6MWD were significantly improved after 3 months and slightly 3 years after LVRS. Gas exchange parameters were significantly improved 3 months after surgery, but returned to the preoperative levels after 3 years. CONCLUSIONS: LVRS may significantly improve pulmonary function in patients with severe COPD indicating for LVRS. Mid-term pulmonary function 3 years after surgery can be decreased to the level at 3 months after surgery. Three years after LVRS, lung volume and pulmonary ventilation function can be significantly improved, but the improvement in gas exchange function was not significant.

Chronotropic Incompetence and its Relation to Exercise Intolerance in Chronic Obstructive Pulmonary Disease.

BACKGROUND: To study the relationship between chronotropic incompetence (CI) and disease severity and to assess the effect of CI on exercise capacity in patients with chronic obstructive pulmonary disease (COPD). MATERIALS AND METHODS: Arterial blood gas analysis, pulmonary function test and cardiopulmonary exercise testing were conducted in 60 patients with stable COPD and 45 healthy volunteers. CI was defined using the chronotropic response index (CRI = (peak heart rate-resting heart rate) / (220-age-resting heart rate). Based on CRI, patients with COPD were divided into the normal chronotropic group (n = 23) and CI group (n = 37). RESULTS: CI was present in 61.7% of the patients with COPD. Exercise capacity (peak oxygen uptake as percentage of predicted value, peak VO2%pred), peak heart rate and CRI were significantly lower in patients with COPD than in controls. However, resting heart rate was significantly higher than in controls. FEV1%pred and exercise capacity were significantly decreased in the CI group when compared with those in the normotropic group. There was significant association between CRI with FEV1%pred and peak VO2%pred. Multivariate regression analysis showed that CRI and FEV1%pred were independent predictors of exercise capacity in patients with COPD. A cutoff of 0.74 for the CRI showed a specificity of 94.1% in predicting patients with a peak VO2%pred < 60%. CONCLUSIONS: CRI was associated with disease severity in patients with COPD. CI may be an important parameter to reflect exercise capacity in patients with COPD.

Humoral immune responses induced by anti-idiotypic antibody fusion protein of 6B11scFv/hGM-CSF in BALB/c mice.

BACKGROUND: We have previously developed and characterized a monoclonal anti-idiotype antibody, designated 6B11, which mimics an ovarian carcinoma associated antigen OC166 - 9 and whose corresponding monoclonal antibody is COC166 - 9 (Ab1). In this study, we evaluate the humoral immune responses induced by the fusion protein 6B11 single-chain variable fragment (scFv)/human granulocyte macrophage colony-stimulating factor (hGM-CSF) and 6B11scFv in BALB/c mice. METHODS: The fusion protein 6B11scFv/hGM-CSF was constructed by fusing a recombinant single-chain variable fragment of 6B11scFv to GM-CSF. BALB/c mice were administrated by 6B11scFv/hGM-CSF and 6B11scFv, respectively. RESULTS: The fusion protein 6B11scFv/hGM-CSF retained binding to the anti-mouse F (ab) 2' and was also biologically active as measured by proliferation of human GM-CSF dependent cell TF1 in vitro. After immunization with the 6B11scFv/hGM-CSF and 6B11ScFv, BALB/c mice showed significantly enhanced Ab3 antibody responses to 6B11scFv/hGM-CSF compared with the 6B11scFv alone. The level of Ab3 was the highest after the first week and maintained for five weeks after the last immunization. Another booster was given when the Ab3 titer descended, and it would reach to the high level in a week. CONCLUSION: The fusion protein 6B11scFv/hGM-CSF can induce humoral immunity against ovarian carcinoma in vivo. We also provide the theoretical foundation for the application of the fusion protein 6B11scFv/hGM-CSF for active immunotherapy of ovarian cancer.

Combined inhaled corticosteroid and long-acting ß2-adrenergic agonist therapy for noncystic fibrosis bronchiectasis with airflow limitation: An observational study.

BACKGROUND AND OBJECTIVE: There is presently no clear evidence on the effect of combined treatment for non-cystic fibrosis (non-CF) bronchiectasis with inhaled corticosteroid (ICS) and long-acting ß2-adrenergic agonist (LABA). The objective of this study is to assess the efficacy and safety of salmeterol-fluticasone combined inhaled therapy for non-CF bronchiectasis with airflow limitation. METHODS: An observational study was performed in 120 non-CF bronchiectasis patients diagnosed by high-resolution computed tomography (HRCT) scanning of the chest. Patients received either routine therapy or salmeterol-fluticasone (100/500 µg daily) combined inhaled therapy on the basis of routine therapy. Clinical symptoms, health-related quality of life (HRQL), lung function, short-acting ß2-adrenergic agonist (SABA) use, and safety were monitored throughout the study. RESULTS: OF the 120 subjects, 60 received combined inhaled therapy and 60 received routine therapy. Compared to the control group, the combined inhaled therapy group showed significant improvement in their clinical symptom scores (-2.21 vs. -0.31, P = 0.002) and a reduction in number of weekly SABA usage (-4.2 vs. 0.1, P < 0.01). In addition, patients in the inhaled therapy group achieved a significant improvement in HRQL based on mMRC (-1.51 vs. -0.31, P < 0.005) and SGRQ (-7.83 vs. -2.16, P < 0.01) scoring accompanied with no severe adverse events. There were fewer exacerbation frequencies in the combined inhaled therapy group over the 12 months of treatment compared to the control group (1 [0-2] vs. 2 [1-4], P = 0.017). Furthermore, stratified analysis indicated that combined inhaled therapy partially improve lung function for patients for whom it is severely impaired and those with pseudomonas aeruginosa isolated. CONCLUSION: Our results show that salmeterol-fluticasone combined inhaled therapy should be effective and safe for non-CF bronchiectasis patients especially for those patients with poor lung function or pseudomonas aeruginosa isolated.

[In vitro study of the antitumor immune responses induced by anti-idiotypic minibody vaccine of ovarian cancer].

OBJECTIVE: To evaluate whether anti-tumor immune response can be induced in vitro with 6B11 anti-idiotypic minibody. and to explore its probability as ovarian cancer vaccine. METHODS: Separated human peripheral blood mononuclear cells (PBMC) were stimulated and cultured by 6B11 minibody. The proliferations of PBMC and cytotoxin were observed by (3)HTdR and (51)Cr release test respectively. ELISA(Enzyme-Linked Immunosorbent Assay) test and Immune Flow Cytometry were used to analyze IFN-gamma in supernatant of the cultured cdlls and the change of T lymphocyte phenotype of PBMC with 6B11 minibody stimulated. RESULTS: 6B11 minibody could stimulate PBMC to proliferate, the best dose was 20 mg/L; it performed cytotoxin function to ovarian carcinoma cell line expressing OC166-9. IFN-gamma maintained at high level after stimulation. It stimulated proliferation of CD3(+) T cell and CD4(+) from PBMC after stimulation respectively. CD8(+) T cell proliferation was not clear. There was significant difference between stimulation and unstimulation in CD4(+)/CD8(+) ratio. CONCLUSION: 6B11 anti-idiotypic minibody can induce both humoral and cellular immunity against ovarian carcinoma in vitro. This paper has provided strong experimental evidence for clinical use of 6B11 minibody as anti-idiotype vaccines against ovarian carcinoma.

[Effect of lung transplantation on pulmonary function in 5 patients with chronic obstructive pulmonary disease].

OBJECTIVE: To investigate the therapeutic effect of lung transplantation on pathophysiology and pulmonary function in chronic obstructive pulmonary disease (COPD) patients. METHODS: Five male COPD (grade IV) patients, aged 51 to 63 yr, were enrolled in the study. The patients underwent pulmonary function tests and the following measurements 2 weeks before and 2 months after the operation. The measured parameters included forced vital capacity (FVC), forced expiratory volume in one second (FEV(1)), FEV(1)/FVC, maximal ventilatory volume (MVV), residual volume (RV), total lung capacity (TLC), RV/TLC, inspiratory capacity (IC), thoracic gas volume (TGV), peak expiratory flow (PEF), total airway resistance (R(aw)total), diffusion capacity for CO of lung (D(L)CO), diffusion capacity for CO of lung/alveolar volume (D(L)CO/V(A)), 6 minute walk distance (6MWD), partial pressure of oxygen in arterial blood (PaO(2)), alveolar-artery oxygen gradient [P((A-a))O(2)], oxygen saturation in arterial blood (SaO(2)), partial pressure of carbon dioxide in arterial blood (PaCO(2)) and mean pulmonary arterial pressure (mPAP). RESULTS: The measured parameters before vs after the operation were as follows: MVV (23.6 +/- 5.8) vs (71.6 +/- 21.8) L, FEV(1) (0.68 +/- 0.21) vs (1.85 +/- 0.46) L, FEV(1)/FVC (37.4 +/- 8.3)% vs (75.6 +/- 13.9)%, PaO(2) (60.0 +/- 9.1) vs (86.2 +/- 2.9) mm Hg (1 mm Hg = 0.133 kPa), SaO(2) (90.0 +/- 4.6)% vs (96.8 +/- 0.5)% and mPAP (31.2 +/- 5.5) vs (16.6 +/- 1.8) mm Hg; all were significantly improved in the 5 cases (all P < 0.05); IC [(1.16 +/- 0.26) vs (1.83 +/- 0.35) L], TGV [(6.52 +/- 0.27) vs (4.52 +/- 0.29) L], RV [(5.12 +/- 0.39) vs (3.20 +/- 0.32) L], RV/TLC [(71.0 +/- 5.6)% vs (51.3 +/- 2.5)%] and R(aw) total [(6.62 +/- 0.99) vs (2.48 +/- 0.87) cm H2O.L(-1).s(-1)] were significantly improved in 3 of the 5 patients (all P < 0.05); PEF [(1.65 +/- 0.40) vs (3.92 +/- 1.63) L/s], D(L)CO [(8.5 +/- 3.0) vs (21.0 +/- 6.2) ml.min(-1).mm Hg(-1)] and 6MWD [(46.8 +/- 14.7) vs (246.8 +/- 51.9) m] were significantly increased in 4 of the 5 patients (all P < 0.05). FVC [(1.85 +/- 0.40) vs (2.45 +/- 0.49) L], TLC [(7.19 +/- 0.15) vs (6.26 +/- 0.73) L], D(L)CO/V(A) [(2.90 +/- 1.50) vs (5.41 +/- 0.87) L.min(-1).mm Hg(-1)], P((A-a))O(2) [(37.6 +/- 16.3) vs (17.8 +/- 6.3) mm Hg] and PaCO(2) [(44.6 +/- 7.7) vs (37.4 +/- 3.4) mm Hg] were also improved but did not reach significance (all P > 0.05). CONCLUSION: Spirometry, airway resistance, residual capacity, diffusion capacity, exercise tolerance and gas exchange were improved remarkably after lung transplantation in COPD patients.

Change in pulmonary function in chronic obstructive pulmonary disease stage 0 patients.

This study aims to elucidate the change in pulmonary function in stage 0 COPD patients. A total of 48 stage 0 COPD patients and 46 healthy adults were included in the study. The status of their pulmonary function was determined by an impulse oscillometry (IOS) system, and the spirometric indexes such as forced vital capacity, maximum expiratory flow-volume (MEFV) curve, total respiratory impedance (Zrs) and respiratory resistance (Rrs) between the two groups were compared. Significant decreases in the values of forced expiratory flow (FEF) at both 75% and 50% of the vital capacity of the predicted value (EF75/pre and FEF50/pre) were detected in stage 0 COPD patients compared with those in the control (P < 0.05). Significant increases were found in the resonant frequency (Fres) (14.37±3.63 VS 11.26±2.61), total respiratory impedance (Z5) compared with the prediction (Z5/pre) (135.65±19.37 VS 104.69±20.64), total airway resistance at 5 Hz (R5) compared with prediction (R5/pre) (128.46±20.14 VS 100.60±20.98) and peripheral airway resistance (R5-R20) compared with prediction (R5-R20/pre) (282.34±192.83 VS 109.31±80.05) in the study group compared with those in the control(all P < 0.05). The reactance at 5 Hz (X5) (-0.14±0.05 VS -0.08±0.05) in the stage 0 COPD group was markedly lower than that in the healthy group (P < 0.05). Disturbance in the small airway may be detected by the MEFV curve and IOS, and these indexes would be valuable in diagnosing stage 0 COPD.

[Immuno-therapeutic study of anti-idiotype minibody (single chain Fv-CH3) on ovarian carcinoma bearing mice].

OBJECTIVE: An anti-idiotypic minibody with optimal antigenicity which mimicking ovarian cancer antigen was used for therapeutic research in mice model bearing ovarian cancer. METHODS: Using gene engineering technique, prokaryotic expression vector was constructed by genetic fusion of 6B11scFv to human IgG1 hinge and CH3 region. The fusion protein named minibody was induced with IPTG in E. coli and analyzed with Western blot and inhibition ELISA tests respectively. Twenty human-PBL-SCID mice bearing i.p. Skov3.ip1 cells were divided into two groups (10 per-group), 10 mice were immunized repeatedly by minibody every two weeks for three times. Indirect ELISA test was employed for analyzing the characterization of anti-anti-idiotypic scFv (Ab3). The latent period of ascites growth and the mean survival time were observed respectively. CD4+ and CD8+ T cells from the spleen of immunized mice were assayed by flow cytometry. RESULTS: SDS-PAGE gel electrophoresis showed that a protein band with molecular weight of 50,000 appeared as the expected size after transformation and induction the host bacteria BL21 (DE3). The expressed minibody could be reacted with COC166-9 (Ab1 of 6B11) and binding goat anti-human IgG1 antibody in Western blot. Inhibition ELISA showed minibody had the capacity of binding ovarian cancer monoclonal antibody COC166-9 instead of primal antigen. Ab3 could be detected in the sera of immunized mice with minibody by ELISA test. Ab3 reached the highest at the 14th day after last vaccination and lasted for 6 weeks. The ratio of CD4+/CD8+ was the highest at the 13th day after last vaccination. The latent period of ascites growth were (37.7 +/- 5.5) days and (48.6 +/- 14.3) days (P = 0.04) respectively; while the mean survival time were (42.5 +/- 1.8) days and (59.4 +/- 16.8) days (P = 0.011) in the control and minibody group respectively. CONCLUSIONS: These results demonstrate the successful construction and expression minibody with good immune activities of 6B11scFv and human IgG1 molecules function. Antigenicity is increased without adjuvants and partial humanization is realized. Minibody can induce humoral anti-idiotypic immunity responses against ovarian carcinoma in vivo. When ascites formation was delayed or prevented and the survival was prolonged in minibody group. We expect that minibody may be used as tumor vaccine to ovarian carcinoma in the future clinical trails.

Evaluation of respiratory impedance in patients with chronic obstructive pulmonary disease by an impulse oscillation system.

An impulse oscillometry system (IOS) assesses pulmonary resistance and reactance. The present study investigated which IOS measurement is correlated with airflow obstruction, airway conductance and lung volume in chronic obstructive pulmonary disease (COPD). A total of 180 patients with COPD were selected and 95 agreed to follow­up 1 year after the initial tests. IOS measurements [R5, R20, X5 and resonant frequency (Fres)], body plethysmography [forced end­expiratory flow (FEF)75, total lung capacity, residual volume (RV) and total inspiratory resistance (Rtot)] and spirometry [forced expiratory volume in 1 sec (FEV1)] were performed. Pearson's or Spearman's correlation tests were used to determine the correlation between the IOS and other measurements. R5, X5 and Fres were all significantly associated (P<0.05) with FEV1, FEF75, RV and Rtot. However, R20 was not correlated with these measurements except from FEF75 and Rtot (r values were all<0.25). The strongest associations were observed with FEV1 and the reactance measurements X5 (r=0.635), Fres (r=­0.721) and R5 (r=­0.496); FEF75 with X5 (r=0.505), Fres (r=­0.629) and R5 (r=­0.468); RV with X5 (r=­0.485), Fres (r=0.570) and R5 (r=0.326); and Rtot with X5 (r=­0.691), Fres (r=0.632) and R5 (r=0.570). There was a significant increase in FEV1 and X5 after one year as compared with the RV. The other measurements did not change over the year. The changes in X5, Fres and R5 were significantly correlated with the changes in FEV1 and the correlation coefficients were 0.355, ­0.364 and ­0.381, respectively. Similarly, the changes in X5, Fres and R5 were significantly correlated with the changes in RV and the correlation coefficients were ­0.264, 0.287 and 0.318, respectively. In the COPD patients, the IOS reactance measurements were more closely correlated with other pulmonary function measurements rather than with resistance measurements. The IOS reactance measurements, particularly X5, appear to indicate changes in pulmonary compliance caused by airflow obstruction.