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Objective To use the liquid phase coprecipitation method for preparing gemcitabine magnetic nanoparticles to the pouch,and explore the preparation process in a number of conditions. Methods The effect of different stirring speeds,Fe3+/Fe2+ratio,pH,temperature of Fe3O4 powder and emulsification condition,ultrasonic time,curing temperature on the preparation of target?ed magnetic nanoparticles were observed. Results The preparation of Fe3O4 nanoparticles was as follows:800 r/ min of stirring speed,1.7∶1 Fe3+/Fe2+ratio,pH=9,the reaction temperature of 60℃,and the 5∶40 water/oil compared,ultrasonic time of 10 min, 100℃curing temperature. Conclusion Fe3O4 powder with small particle size,high purity,and no agglomeration are prepared,the stability of the gemcitabine magnetic nanoparticles capsule is good.
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Objective To investigate the relationship among peroxisome proliferators - activated receptor gamma (PPAR-γ), pul-monary arterial systolic pressure(PASP) ,and insulin resistance in Chronic Obstructive Pulmonary Disease (COPD) patients. Methods A-mong 63 COPD patients, 30 patients with level of PASP above 40mmHg were enrolled in PAH group and other 33 patients were enrolled in COPD group. Twenty healthy medical examination subjects were enrolled in control group. The expression of PPAR-γ, mRNA was detected by real time fluorescent quantitative RT- PCR. Radioimmunoassay was used to measure the level of fasting plasma insulin (FIN). Fasting plas-ma glucose (FPG) was detected by glucose oxidase method. Results The expression of PPAR-γ mRNA was significantly decreased in PAH and COPD group, while FPG, FIN and IRI increased significantly. PAH group had more increased PASP, decreased expression of PPAR-γ and higher IRI than COPD group. Expression of PPAR-γ was negatively related to PASP and IRI. Conclusions The significantly down reg-ulated expression of PPAR-γ maybe explain the higher FPG and PASP.
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Objective To observe the effects of CD4~+ CD25~+ regulatory T cells ( CD4~+ CD25~+ Treg) on the airway inflammation of asthmatic rats. Methods CD4~+ CD25~+ Treg of OVA- immune tolerance rats were transferred to asthmatic rats. Then bronchoalveolar lavage fluid (BALF) was collected, and cytology study was conducted. The IL-4, IL-5, IFN-γ and OVA-specific serum IgE level in BALF were determined by ELISA. The lung tissue was obtained, and histologieal analysis was done through H. E. Results Total cells number, the percentage of lymphocytes and neutrophils in BALF, the IL-4 and IL-5 BALF levels and the OVA-specific serum IgE level of adoptive transfer group were decreased ( P < 0.05 ) , and the percentage of eosinophils ( Eos) was significantly lower than that of asthma group ( P < 0.01) , while its BALF IFN-γ level was higher than that of asthma group( P <0. 05). Compared with that of asthma group, peribronchiole inflammatory of treated group was alleviated. Conclusion CD4 ~+ CD25~+ Treg of OVA- immune tolerance rats transferred to asthmatic rats can significantly alleviate the airway inflammation of asthmatic rats.
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Objective To investigate dexamethasone on airway inflammation and CD4+ CD25 + regulatory T cells (CD4+ CD25 +Tr) of asthmatic rats,and elucidate the possible mechanism of dexamethasone in treatment of asthma.Methods 30 Wistar rats were randomly divided into control group,asthma group and dexamethasone-treated group.Bronchoalveolar lavage fluid (BALF) was collected,and cytology study was conducted.The lung tissue was obtained and pathologic analysis was done through HE stain.Flow eytometry was used to detect the CD4+ CD25 +Tr ratio in PBMCs.Results Total cells number,the percentage of lymphocytes,neutrophils and eosinophils (Eos)in BALF of dexamethasone-treated group were lower than that of asthma group (P＜0.05,P＜0.01).Compared with the asthma group,less infiltration of inflammatory cells in lung tissues was observed in the dexamethasone-treated group.CD4+ CD25 + Tr of asthma group was lower than that of control and dexamethasone-treated group (P＜0.05).Conclusion Dexamethasone could suppress airway inflammation of asthmatic rats,which probably be due to increasing the number of CD4+ CD25 + Tr.
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Objective To investigate the expression of CD40 and CD40L on the surface of peripheral blood mononuclear cells(PBMCs)in asthmatic rats and the effect of anti-CD40L McAb on cytokines of it.Methods Flow cytometry and RT-PCR were used to detect the expression of CD40 and CD40L of PBMCs ih asthmatic rats.After the PBMCs Was treated with anti.CIMOL McAb.ELISA was used to detect the levels of IL-4 and IFN-γin the supematants of cultured cells.Results Compared with the normal control group.the expression of CD40 and CD40L of PBMCs in asthImatic rats increased(P＜0.05).Compared with the untreated group,the level of IL-4 and the ratio of IL4/IFN-γ decreased after the PBMCs were treated with anti-CD40L McAb(P＜0.05).Conclusion The expression of CD40 and CD40L on the surface of PBMCs in asthmatic rats Was unregulated.Anti-CD40L Mcab Can decrease the level of IL-4 and the ratio of IL_4/IFN-γ.
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Objective To investigate the immunological mechanism of inhibitory effect of Danshen injection combined with dexamethasone(DXM) on asthmatic airway inflammation.Methods 50 Wistar rats were randomly divided into normal control(NC),asthma,Danshen,DXM and Danshen+DXM group.Cytology study of Bronchoalveolar lavage fluid(BALF) was conducted.Pathology of lung tissue was done through HE.Flow eytometry was used to detect CD4+CD25+ regulatory T Cells(CD4+CD25+ Treg) ratio in peripheral blood mononuclear cells(PBMCs).IL-4 and IL-5 levels in BALF were detected by ELISA.Results Total cells number,percentage of lymphocytes,neutrophils and eosinophils(Eos) in BALF of the three treated groups were lower than that in asthma group(P<0.05,P<0.01),particularly in Danshen+DXM group,which showed significant difference as compared with the other two treated groups(P<0.05).There was severe inflammation in lung tissue of asthma group,moderate inflammation in Danshen group and DXM group,and no inflammation of Danshen+DXM group.CD4+CD25+ Treg/CD4+ T ratio in the three treated groups were higher than that in asthma group,and the levels of IL-4 and IL-5 were lower than those in asthma group(P<0.05).In Dansben+DXM group,it showed significant difference on the change of CD4+CD25+ Treg,IL-4 and IL-5 as compared with other treated groups(P<0.05).Conclision Danshen injection combined with DXM could suppress airway inflammation in asthmatic rats,which may be through increasing the expression of CD4+CD25+ Treg,decreasing the levels of IL-4 and IL-5 and resuming the balance of Th1/Th2.
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Objective To investigate the molecular mechanism of inhibitory effect of Danshen injection on allergic airway inflamma-tion of asthma. Methods 30 SD rats were random divided into control group, asthma group and Danshen injection group. Bronchoalveolar lavage fluid (BALF) was collected, and cytology study was conducted. The lung tissue was obtained and pathologic analysis was done through HE staining, lnterleukin-13 (IL-13) and Eotaxin in lung tissue were measured by RT - PCR and immunohistochemistry SP method. Results Compared with the asthma group, less infiltration of inflammatory cells in lung tissues was observed in the Danshen injection group. Total cell number, the percentage of lymphocytes, neutrophils and eosinophils (Eos) in BALF of Danshen injection group were lower than that of asthma group (P<0.05, P<0.01). The expression of IL-13 and eotaxin in lung tissue of asthma group was higher than that of control group and Danshen injection group (P<0.05). The expression of IL-13 was negatively correlated with eotaxin (r=0.90, P< 0.05). Conclusion Danshen injection could suppress airway inflammation of asthmatic rats, which probably be through decreasing the ex-pression of IL-13 and eotaxin in the lung tissue of asthmatic rats.
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In order to investigate the clinical value of vascular endothelial growth factor (VEGF) combined with interferon-γ (IFN-γ) in diagnosing malignant pleural effusion and tuberculous plearal effusion, 42 cases of malignant pleurai effusion and 45 cases of tuberculous plcural effusion in Tongji Hospital, from March 2004 to May 2005, were included. The carcinoembryonic antigen (CEA), VEGF and IFN-γ levels of pleural effusion were detected by using ELISA, and adenosine deaminasc (ADA) activity was determined by using enzyme kinetic analytical method. The sensitivity, specific-ity, accuracy and area under the curve (AUCROC) of CEA and VEGF, VEGF/IFN-γ ratio, ADA and IFN-γ were measured by receiver operating characteristic curve (ROC). The results showed that CEA, VEGF levels and VEGF/IFN-γ ratio were significantly higher and the ADA and IFN-γ levels were significantly lower in malignant group than those in tuberculous group (P<0.01). The sensitivity, specificity, accuracy and AUCROC of VEGF/IFN-γ ratio (88.7%, 99.8%, 94.4%, 0.96 respectively) were higher than those of CEA (67.8%, 96.1%, 82.4%, 0.78 respectively) and VEGF (81.5%, 84.3%,82.9%, 0.79 respectively). The sensitivity, specificity, accuracy and AUCROC of IFN-γ (85.7%, 96.4%,90.9%, 0.94 respectively) were higher than those of ADA (80.2%, 87.6%, 83.8%, 0.81 respectively).It was concluded that VEGF/IFN-γ ratio and IFN-γ could be used as valuable parameters for the dif-ferential diagnosis of malignant pleural effusion and tuberculous pleural effusion.
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The changes of CD4+CD25+ regulatory T cells (CD4+CD25+ Treg) and Foxp3 mRNA in peripheral blood mononuclear cells (PBMCs) from patients with asthma were investigated in order to elucidate the possible roles of CD4+CD25+ Treg in the development of asthma. The peripheral blood samples were collected from 29 healthy controls (normal control group) and 78 patients with asthma which included 30 patients in exacerbation group, 25 patients in persistent group, and 23 patients in remission group. By using flow cytometry and RT-PCR, the CD4+CD25+ Treg ratio and Foxp3 mRNA in PBMCs were detected. The CD4+CD25+ Treg ratio and Foxp3 mRNA in PBMCs of exacerbation and persistent groups were lower than that of remission and normal control groups (P＜0.05). Although the CD4+CD25+ Treg ratio and Foxp3 mRNA of remission group were also lower than that of normal control group, there was no significant difference between them (P＞0.05). As compared with persistent group, exacerbation group had lower CD4+CD25+ Treg ratio and Foxp3 mRNA (P＜0.05). It was indicated that the decrease of CD4+CD25+Treg ratio and its function in PBMCs may be responsible for pathogenesis of asthma.
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The changes of CD4(+)CD25(+) regulatory T cells (CD4(+)CD25(+) Treg) and Foxp3 mRNA in peripheral blood mononuclear cells (PBMCs) from patients with asthma were investigated in order to elucidate the possible roles of CD4(+)CD25(+) Treg in the development of asthma. The peripheral blood samples were collected from 29 healthy controls (normal control group) and 78 patients with asthma which included 30 patients in exacerbation group, 25 patients in persistent group, and 23 patients in remission group. By using flow cytometry and RT-PCR, the CD4(+)CD25(+) Treg ratio and Foxp3 mRNA in PBMCs were detected. The CD4(+)CD25(+) Treg ratio and Foxp3 mRNA in PBMCs of exacerbation and persistent groups were lower than that of remission and normal control group (P0.05). As compared with persistent group, exacerbation group had lower CD4(+)CD25(+) Treg ratio and Foxp3 mRNA (P<0.05). It was indicated that the decrease of CD4(+)CD25(+) Treg ratio and its function in PBMCs may be responsible for pathogenesis of asthma.
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The expression of resistin protein in normal human abdominal, thigh, pregnant women abdominal, non-pregnant women abdominal subcutaneous adipose tissue and placenta and the relationship between obesity, type 2 diabetes mellitus (T2DM), pregnant physiological insulin resistance (IR) and gestational diabetes mellitus (GDM) was investigated. The expression of resistin protein in normal human abdominal, thigh, pregnant women abdominal, non-pregnant women abdominal subcutaneous adipose tissue and placenta was detected by using Western blotting method. Fasting serum glucose concentration was measured by glucose oxidase assay. Serum cholesterol (CHOL), serum triglycerides (TG), serum HDL cholesterol (HDL-C) and serum LDL cholesterol (LDL-C) were determined by full automatic biochemical instrument. Fasting insulin was measured by enzyme immunoassay to calculate insulin resistance index (IRI). Height, weight, systolic blood pressure (SBP) and diastolic blood pressure (DBP) were measured to calculate body mass index (BMI) and body fat percentage (BF %). Resistin protein expression in pregnant women placental tissue (67 905 +/- 8441) (arbitrary A values) was much higher than that in subcutaneous adipose tissue in pregnant women abdomen (40 718 +/- 3818, P < 0.01), non-pregnant women abdomen (38 288 +/- 2084, P < 0.01), normal human abdomen (39 421 +/- 6087, P < 0.01) and thigh (14 942 +/- 6706, P < 0.001) respectively. The resistin expression in abdominal subcutaneous adipose tissue showed no significant difference among pregnant, non-pregnant women and normal human, but much higher than that in thigh subcutaneous adipose tissue (P < 0.001). Pearson analysis revealed that resistin protein was correlated with BMI (r = 0.42), fasting insulin concentration (r = 0.38), IRI (r = 0.34), BF % (r = 0.43) and fasting glucose (r = 0.39), but not with blood pressure, CHOL, TG, HDL-C and LDL-C. It was suggested that resistin protein expression in human abdominal subcutaneous adipose tissue was much higher than that in human thigh subcutaneous adipose tissue. Resistin was closely related with central obesity, leading to IR, subsequently obesity and T2DM. Resistin protein expression in placental tissue was much higher than that in subcutaneous adipose tissue in normal human abdomen, pregnant abdomen, non-pregnant women abdomen and thigh. It was indicated that resistin protein could be secreted from human placental tissue. Resistin might be one of the factors that lead to pregnant physiological IR and GDM.
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The expression of resistin protein in normal human abdominal, thigh, pregnant women abdominal, non-pregnant women abdominal subcutaneous adipose tissue and placenta and the relationship between obesity, type 2 diabetes mellitus (T2DM), pregnant physiological insulin resistance (IR) and gestational diabetes mellitus (GDM) was investigated. The expression of resistin protein in normal human abdominal, thigh, pregnant women abdominal, non-pregnant women abdominal subcutaneous adipose tissue and placenta was detected by using Western blotting method.Fasting serum glucose concentration was measured by glucose oxidase assay. Serum cholesterol (CHOL), serum triglycerides (TG), serum HDL cholesterol (HDL-C) and serum LDL cholesterol (LDL-C) were determined by full automatic biochemical instrument. Fasting insulin was measured by enzyme immunoassay to calculate insulin resistance index (IRI). Height, weight, systolic blood pressure (SBP) and diastolic blood pressure (DBP) were measured to calculate body mass index (BMI) and body fat percentage (BF %). Resistin protein expression in pregnant women placental tissue (67 905±8441) (arbitrary A values) was much higher than that in subcutaneous adipose tissue in pregnant women abdomen (40 718 ± 3818, P ＜ 0.01), non-pregnant women abdomen (38 288±2084, P＜0.01), normal human abdomen (39 421±6087, P＜0.01)and thigh (14 942 ±6706, P＜0. 001) respectively. The resistin expression in abdominal subcutaneous adipose tissue showed no significant difference among pregnant, non-pregnant women and normal human, but much higher than that in thigh subcutaneous adipose tissue (P＜0. 001). Pearson analysis revealed that resistin protein was correlated with BMI (r=0.42), fasting insulin concentration (r=0.38),IRI (r=0. 34), BF % (r=0.43) and fasting glucose (r=0. 39), but not with blood pressure,CHOL, TG, HDL-C and LDL-C. It was suggested that resistin protein expression in human abdominal subcutaneous adipose tissue was much higher than that in human thigh subcutaneous adipose tissue. Resistin was closely related with central obesity, leading to IR, subsequently obesity and T2DM. Resistin protein expression in placental tissue was much higher than that in subcutaneous adipose tissue in normal human abdomen, pregnant abdomen, non-pregnant women abdomen and thigh. It was indicated that resistin protein could be secreted from human placental tissue. Resistin might be one of the factors that lead to pregnant physiological IR and GDM.
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Objective To investigate changes of CD~+_4CD~+_ 25 regulatory T cells (CD~+_4CD~+_ 25 Treg) and forkhead/winged helix transcription factor(Foxp3) mRNA in peripheral blood mononuclear cells (PBMCs) from patients with asthma, so as to elucidate the possible roles of CD~+_4CD~+_ 25 Treg in the development of asthma. Methods The peripheral blood samples were collected from 29 healthy controls (normal control group) and 78 patients with asthma which included 30 patients in exacerbation group, 25 patients in persistent group, and 23 patients in remission group. By using flow cytometry and RT-PCR, the CD~+_4CD~+_ 25 Treg ratio and Foxp3 mRNA in PBMCs were detected. Results The CD~+_4CD~+_ 25 Treg ratio and Foxp3 mRNA in PBMCs of exacerbation and persistent group were lower than that of remission group and normal control group (P0.05). Compared with persistent group, exacerbation group had lower CD~+_4CD~+_ 25 Treg ratio and Foxp3 mRNA (P
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Objective To investigate the efficacy and adverse effects of methotrexate(MTX) and Bacille Calmette Guerin-polysaccharide nucleic acid(BCG-PSN) in the treatment of steroid dependent asthma.Methods Sixty-eight patients with steroid dependent asthma were divided into therapy group and control group.The patients in therapy group were treated with MTX and BCG-PSN,while the control group was given only MTX.The dose of oral steroid daily,symptom scores,peak flow rates(PEF) and expiratory volume in one second(FEV1) were monitored.Results Compared with before treatment,daily steroid dosage and symptom scores of two groups significantly decreased(P0.05),while PEF and FEV1 significantly increased in therapy group(P0.05).Conclusion The combined use of MTX and BCG-PSN may be more beneficial than the single use of MTX for the treatment of steroid-dependent asthma.
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Aim To investigate the inhibitive effects of trichostatin A(TSA) on telomerase activity of HL-60 cells and expression of subunit hTERT during apoptosis in vitro and its mechanism.Methods The proliferative activity of HL-60 cells was assessed using morphology and MTT assay.Cell apoptosis was confirmed using Flow Cytometer.Telomerase activity was examined using TRAP-ELISA.The expression status of telomerase subunits was analyzed using RT-PCR.Results A time-and dose-dependent inhibition was detected in HL-60 cells treated with TSA.After 48 h TSA(300 nmol?L~(-1)) treatment,the apoptotic rate detected using cytometric assay(Annexin V/PI double staining)of HL-60 cells was 42.6%.Telomerase activity and expression level of hTERT and the key subunit of telomerase decreased at 24-hour after TSA treatment.No significant changes were observed in the expression of hTR,hTP and the other two subunits of telomerase.Conclusion TSA inhibits telomerase activity and induces apoptosis in HL-60 cells.The underlying mechanism might be related to the down regulation of hTERT transcription.
