Objective To investigate correlation of pulmonary heart disease (PHD)with the diaphragmatic muscle fatigue(DMF). Methods Impedance Pneumoplethysmogram(IPG) and Impedance respirogram(IRG) were detected in 64 cases of patients with PHD,and investigate DMF rheogram positive rate,also analysed various parameters of IPG in positive and negative groups. Results (1)The positive rate of DMF.Rheogram in PHD with and without respiratory failure are 75％ and 21.9％ respectively.(2)The positve rate in PHD with and without heart failue are 80％ and 46.3％ respectively.(3)The mean values of various parameters of IPG in positive and negative DMF rheogram have no significant difference.Conclusion Respiratory failure and heart falure in PHD are closely correlated with DMF. However,the occurance of DMF are not parallel with the severity degree of PHD.

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.

To establish a better method of primary culture for alveolar epithelial type II cells (AEC II) and to study its bionomics, alveolar epithelial type II cells were isolated by digestion with trypsin and collagenase, which were then purified by plated into culture flask coated with rat immunoglobulin G. The purified AEC II were identified by alkaline phosphatase staining, electron microscopy, immunocytochemical staining of pulmonary surfactant protein A (SPA). The SPA expression and transfection characteristics were compared with those of A549 cell line. The results showed that AEC II could be isolated by digestion with trysin and collagenase and purified by adhesive purification by using IgG, with a yield of about 2-3 x 10(7), and a purity of about 75%-84%. Cells could be quickly identified with AKP staining. AEC II were different from A549 cell line in terms of SPA expression and transfection characteristics. It is concluded that adhesive purification with IgG can improve the purity of AEC II, and AKP staining is simple in cell identification. AEC II can not be completely replaced by A549 cells in some studies because the differences between them, such as SPA expression.
Cell Culture Techniques/*methods ; Cell Separation/*methods ; Cells, Cultured ; Ecology ; Epithelial Cells/*cytology ; Immunoglobulin G/pharmacology ; Pulmonary Alveoli/*cytology ; Pulmonary Surfactant-Associated Protein A/biosynthesis ; Rats, Wistar

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

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 (P<0.05). Although the CD4(+)CD25(+) Treg ratio and Foxp3 mRNA of remission group 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.

AIM:To investigate the effect of human Kv1.5 gene transfection on the proliferation and apoptosis in human airway smooth muscle cells(HASMCs).METHODS:Kv1.5 gene was transiently transfected into HASMCs with Lipofectamine 2000 and the level of Kv1.5 protein was observed by Western blotting.Ca 2+ concentration in HASMCs was investigated by fluorescent quantitation using fluorospectrophotometer.Flow-cytometry,MTT method and TUNEL were used to detect the effects of Kv1.5 gene transfection on proliferation and apoptosis in HASMCs.RESULTS:(1)Western blotting showed that transfection of human Kv1.5 plasmid significantly increased the Kv1.5 protein expression compared to control group(P

AIM:To investigate the activity change of voltage-dependent delayed rectifier potassium channel(Kv) on human airway smooth muscle cells(HASMCs) after transfection of Kv1.5 antisense oligonucleotides(AsOND),and to discuss the regulating mechanism of Kv1.5.METHODS: The mRNA and protein expressions of Kv1.5 in liposome-mediated Kv1.5 AsOND transfected HASMCs were measured with techniques of reverse transcriptase-polymerase chain reaction(RT-PCR) and Western blotting.Kv activities in transfected HASMCs were investigated with whole-cell patch clamp.RESULTS: After HASMC were transfected by liposome-mediated Kv1.5 AsOND,the mRNA and protein expressions of Kv1.5 were decreased,and Kv activity was inhibited,which made the cell membrane potential(Em) inclined to depolarization.CONCLUSION: Transfection of Kv1.5 AsOND made the function of Kv in HASMCs decreased.Kv1.5 may play a critical role in the regulation of Kv activity.

AIM: To investigate whether Ligustrazine(LTZ) has an effect on the changes of protein kinase C(PKC) signaling pathway induced by inflammatory mediators involved in asthma in normal human peripheral blood lymphocytes (PBL). METHODS: 10 mL peripheral venous blood was obtained from each of 63 health humans and treated as follows. The activities of PKC from cytosolic and membrane fractions in PBL were measured by -ATP-catalyzing assay, after PBL had been isolated and performed by following processes: (1) First: three groups treated with 5 g/L LTZ(n=6) or 5 ?mol/L Ro31-8220 (n=6); Paired untreated PBL served as control of this group, as well as the negative controls of the following groups(n=6); (2)Second : three groups treated with 100 nmol/L Methacholine (Mch, n=5), 5 g/L LTZ+100 nmol/L Mch(n=5)or 5 ?mol/L Ro31-8220(a PKC inhibitor)+100 nmol/L Mch(n=5); (3)Third: three groups treated with 100 nmol/L histamine, 5 g/L LTZ+100 nmol/L histamine(n=5) or 5 ?mol/L Ro31-8220+100 nmol/L histamine(n=5); (4)Fourth: three groups treated respectively with 100 nmol/L PMA(a PKC activator, n=5), 5 g/L LTZ+100 nmol/L PMA(n=5) or 5 ?mol/L Ro31-8220+100 nmol/L PMA(n=5). RESULTS: (1)LTZ had no effect on the activities of PKC in inactive PBL in normal humans; (2) Methacholine or histamine resulted in an increase in membrane PKC activity of normal human PBL, which was partly suppressed by LTZ (all P

0.05),but the contraction of the bronchial rings caused by histamine,potassium chloride,intracellular Ca2+ release and extracellular Ca2+ influx was signifi cantly inhibited by Shenmai injection(compare with those of control,P

In this study, whole cell patch clamp recording technique was employed to investigate the effect of Shenmai Injection (SMI) on L-type calcium current of diaphragmatic muscle in rats. The result showed that when the diaphragmatic muscle cell was held at -80 mV and depolarized to +60 mV, 10 microl/ml, 50 microl/ml and 100 microl/ml SMI enhanced the inner peak L-type calcium current from -(6.8 +/- 0.7) pA/pF (n=7) to -(7.3 +/- 0.8) pA/pF (P>0.05, n=7), -(8.6 +/- 1.0) pA/pF (P<0.05, n=7) and -(9.4 +/- 1.2) pA/pF (P<0.05, n=7), respectively, The rates of L-type calcium current were increased by (7.34 +/- 2.37)%, (25.72 +/- 5.94)%, and (38.16 +/- 7.33)%, respectively. However, it had no significant effect on maximal activation potential and reversal potential. Our results suggested that SMI could activate the calcium channel of the diaphragmatic fibers of the rats, increase the influx of Ca2+, and enhance the contractility of diaphragmatic muscles.
Calcium/metabolism ; Calcium Channels, L-Type/*drug effects ; Diaphragm/drug effects ; Diaphragm/*metabolism ; Drug Combinations ; Drugs, Chinese Herbal ; Patch-Clamp Techniques ; Plant Extracts/*pharmacology ; Rats, Wistar