[Adult hospital acquired pneumonia: a multicenter study on microbiology and clinical characteristics of patients from 9 Chinese cities].

OBJECTIVE: To investigate the etiology and clinical characteristics of hospital-acquired pneumonia (HAP) in China and to provide evidence for appropriate therapy. METHODS: We performed a prospective multicenter study in 13 Chinese urban tertiary hospitals. All HAP cases diagnosed at respiratory general ward and respiratory intensive care unit (RICU) from August 2008 to December 2010 were studied. Epidemiological data, etiology and clinical characteristics of enrolled patients were collected. Sputum or tracheal aspirate and blood cultures, Legionella antibodies and Streptococcus pneumoniae urinary antigen tests were performed. Bacteria to antimicrobial susceptibility test was performed. RESULTS: A total of 610 cases of HAP were diagnosed during the study, with an overall incidence of 1.4% among 42 877 hospitalized patients, while the incidence was 0.9% (362/41 261) in respiratory general ward and 15.4% (248/1616) in RICU. 93.9% (573 cases) of patients had at least one underlying disease, and 91.0% (555 cases) had exposure to at least one antimicrobial agent within 90 days prior to HAP diagnosis. Pathogens were identified in 487 patients, with Acinetobacter baumannii [30.0% (183/610)], Pseudomonas aeruginosa [22.0% (134/610)], Staphylococcus aureus [13.4% (82/610)] and Klebsiella pneumonia [9.7% (59/610)] being the most common pathogens. Eighteen patients (3.0%) had infection with fastidious bacteria. A. baumannii and S. aureus were the more frequent pathogens in the ventilator-associated pneumonia (VAP) cases [50.5% (97/192) and 21.4% (41/192)] as compared to non-VAP cases [20.6% (86/418) and 9.8% (41/418), P < 0.01]. A. baumannii and S. aureus were also frequent pathogens in cases with a score of more than 20 by the acute physiology and chronic health evaluation II (APACHEII) scoring [45.7% (69/151) and 20.5% (31/151)], as compared to cases with a score of less than 20 of APACHE II [24.8% (114/459) and 11.1% (51/459), P < 0.01]. A. baumannii showed high resistance rates to carbapenems [more than 70% (109/142)], and the susceptibility to cefoperazone/sulbactam, polymyxin B and tigecycline were 40.8% (58/142), 99.3% (141/142) and 95.8% (136/142) respectively. Resistance rates of P. aeruginosa to meropenem and imipenem were 48.8% (40/82) and 70.7% (58/82) respectively. Methicillin-resistant S. aureus (MRSA) accounted for 87.8% (43/49) in all strains of S. aureus. Mortality rate of VAP cases was 34.5% (61/177), significantly more than that of HAP patients [22.3% (135/605), P < 0.05]. The average hospital stay of patients with HAP was (23.8 ± 20.5) days, significantly more than that of the average for inpatients [(13.2 ± 13.6) days, P < 0.01] during the study period. Mean costs of HAP were (108 950 ± 116 608) yuan, significantly higher than the average hospital costs of respiratory inpatients (17 999 ± 33 364) yuan. CONCLUSIONS: Among Chinese patients hospitalized in urban tertiary medical centers, HAP incidence and mortality rate were high, which increased the patients' hospital stay and the medical costs. Common pathogens were A. baumannii, P. aeruginosa, S. aureus and K. pneumonia. The common bacteria of HAP in China showed high resistance rates to antibiotics.

[A multicentre retrospective study of pulmonary mycosis clinically proven from 1998 to 2007].

OBJECTIVE: To investigate the pathogens, clinical manifestations, prognosis of and the risk factors for pulmonary mycosis in China. METHODS: All cases of pulmonary mycosis from 16 centers in 10 cities from Jan. 1998 to Dec. 2007 that met the diagnostic criteria were included for clinical, microbiological and radiological analysis. RESULTS: Totally 474 cases of pulmonary mycosis were retrieved. The top 5 pulmonary mycosis was pulmonary aspergillosis (180 cases, 37.9%), pulmonary candidiasis (162 cases, 34.2%), pulmonary cryptococcosis (74 cases, 15.6%), pneumocystis carinii pneumonia (23 cases, 4.8%) and pulmonary mucormycosis (10 cases, 2.1%). The constituent ratio in the last 3 years was similar to that in the former 7 years. The main pathogens of pulmonary candidiasis were Candida albicans (308/474, 65.0%) and Candida tropicalis (57/474, 12.0%), which were sensitive to common azoles. Compared with bacterial pneumonia, pulmonary mycosis showed more symptoms of hemoptysis (147/474, 31.0%) and pleural effusion (95/474, 20.0%), and less radiological specificity. Classical halo sign (4/474, 0.8%) and crescentic sign (17/474, 3.6%) were only shown in several cases of pulmonary mycosis. The most common underlying diseases were tumor (including solid tumor and malignant hematological diseases) (94/474, 19.8%), chronic obstructive pulmonary disease (52/474, 11.0%), pulmonary tuberculosis (50/474, 10.5%) and diabetes (48/474, 10.1%). Compared with the other common pulmonary mycosis, pulmonary cryptococcosis affected younger patients, and more cases were community-acquired, but fewer cases with underlining diseases or compromised immune function, and had a better prognosis. CONCLUSION: The ahead five species of pulmonary mycosis in China were orderly pulmonary aspergillosis, pulmonary candidosis, pulmonary cryptococcosis, pneumocystis carinii pneumonia and pulmonary mucormycosis. The main pathogens of pulmonary candidosis were Candida albicans and Candida tropicalis, which were sensitive to common azoles. Compared with the other common pulmonary mycosis, pulmonary cryptococcosis catch younger patients, had more community-acquired cases, and had better prognosis.

Primary pulmonary coccidioidomycosis in China.

Coccidioidomycosis is a fungal infection endemic to south-west USA, north Mexico and parts of Central and South America. We report here a case of primary pulmonary coccidioidomycosis in a previously healthy 14-year-old boy in China, which is considered a non-endemic country. The patient had non-specific symptoms of pulmonary infection, including fever, non-productive cough and night sweats. Both spherules and endospores of Coccidioides immitis were seen histologically following transbronchial biopsy of a cavitary lesion. The patient was treated with amphotericin B and fluconazole. Follow up 6 months post discharge found that the patient made a good recovery.

Expression of the Th1-specific cell-surface protein Tim-3 increases in a murine model of atopic asthma.

T-cell immunoglobulin- and mucin-domain-containing molecule-3 (Tim-3) is preferentially expressed on Th1-helper type T-cells and functions to repress the Th1-mediated immune response. However, the role of Tim-3 during the inflammatory pathogenesis of asthma remains unclear. This study determines the expression level of Tim-3 in CD4+ T-cells within the peripheral blood and bronchoalveolar lavage fluid (BALF) isolated from a murine model of atopic asthma and explores the potential role of Tim-3 during the inflammatory response. Mice were randomly divided into normal control, asthma day 1, and asthma day 7 groups, and peripheral blood T lymphocytes and BALF cells were collected. The ratio of Tim-3+/CD4+ cells among the total CD4+cell populations from peripheral blood and BALF was determined by flow cytometry, and the expression of the Tim-3 mRNA was determined by Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR). In contrast with the normal control group, the ratio of Tim-3+/CD4+:CD4+ cells and the level of Tim-3 mRNA in both the peripheral blood T lymphocytes and BALF cells among the asthma day 1 and asthma day 7 groups were significantly increased (p < 0.01), and those in the asthma day 7 group were higher than the asthma day 1 group (p < 0.05). There was also a positive correlation between the ratio of Tim-3+/CD4+:CD4+ detected in BALF and that the ratio detected in peripheral blood T lymphocytes (r = 0.84, p < 0.01). Therefore, the expression of Tim-3 is increased in CD4+ T-cells following airway challenge and likely affects asthma-induced inflammation by repressing the Th1-mediated immune response.

Altered surfactant protein A gene expression and protein homeostasis in rats with emphysematous changes.

BACKGROUND: The decrease of surfactant protein (SP) secreted by the alveolar type II cell is one of the important causes of limiting air of pulmonary emphysema. However, the SP-A gene and protein changes in this disease are rarely studied. This study was undertaken to investigate alterations in SP-A gene activity and protein, and to explore their roles in the pathogenesis of emphysematous changes. METHODS: Twenty Wistar rats were divided randomly into a normal control group (n = 10) and a cigarette smoking (CS) + lipopolysaccharide (LPS) group (n = 10). Ultra-structural changes were observed under an electron microscope. The number of cells positive for SP-A was measured by immunohistochemistry. The mRNA expression and protein level of SP-A in the lung tissues were determined by quantitative polymerase chain reaction (qPCR) and Western blot separately. The protein level of SP-A in lavage fluid was determined by Western blot. RESULTS: The number of cells positive for SP-A of the CS + LPS group (0.35 +/- 0.03) was lower than that of the blank control group (0.72 +/- 0.06, P < 0.05). The level of SP-A in the lung tissues of rats in the CS + LPS group (0.2765 +/- 0.0890) was lower than that in the blank control group (0.6875 +/- 0.1578, P < 0.05). The level of SP-A in the lavage fluid of rats in the CS + LPS group (0.8567 +/- 0.1458) was lower than that in the blank control group (1.3541 +/- 0.2475, P < 0.05). The lung tissues of rats in the CS + LPS group showed an approximate increase (0.4-fold) in SP-A mRNA levels relative to beta-actin mRNA (P < 0.05). CONCLUSIONS: The changes of SP-A may be related to emphysematous changes in the lung. And cigarette smoke and LPS alter lung SP-A gene activity and protein homeostasis.

The efficacy and safety of combination salmeterol (50 microg)/fluticasone propionate (500 microg) inhalation twice daily via accuhaler in Chinese patients with COPD.

BACKGROUND: Few studies of the efficacy and safety of therapy with combinations of salmeterol/fluticasone propionate (SFCs) have been conducted in Chinese patients with COPD, and the benefits of combination therapy in nonsmoking patients with COPD are, to our knowledge, not known. STUDY OBJECTIVES: The aims were to establish the efficacy and tolerability of the therapy with SFC (salmeterol, 50 microg/fluticasone, 500 microg, twice daily) in the management of Chinese COPD patients and to investigate the effectiveness of SFC in nonsmokers with COPD. METHODS AND PATIENTS: This was a randomized, double-blind, placebo-controlled, parallel-group, multicenter study. Changes in prebronchodilator and postbronchodilator FEV(1), quality of life determined by the St. George Respiratory Questionnaire (SGRQ) scores, relief bronchodilator use, nighttime awakenings, and frequency of exacerbations of COPD were measured in patients randomized to receive SFC (n = 297) or placebo (n = 148). Never-smokers, former smokers, and current smokers accounted for 11.7%, 66.7%, and 21.6%, respectively, of the study population. RESULTS: After 24 weeks, the mean changes in prebronchodilator and postbronchodilator FEV(1) were 180 mL (95% confidence interval [CI], approximately 91 to 268; p < 0.001) and 65 mL (95% CI, approximately 14 to 115; p = 0.012), respectively, greater for the SFC group than that for the placebo group. The differences in response to treatment were significant (all p < 0.0001) in former or current smokers but not in never-smokers (p > 0.05). The mean improvement in the total SGRQ score for the SFC group was 5.74 (p < 0.01) greater than that for the placebo group. SFC significantly reduced the frequency of nighttime awakenings and the use of relief bronchodilator. The adjusted ratio of exacerbations of COPD for the SFC group relative to the placebo group was 0.61 (95% CI, approximately 0.45 to 0.84; p < 0.01). There were no significant differences between the SFC and placebo groups in safety measures. CONCLUSIONS: SFC therapy achieved sustained improvement in lung function, quality of life, and control of symptoms, and was well tolerated in Chinese patients. Greater improvements in lung function were found only for COPD patients with a history of smoking. TRIAL REGISTRATION: http://ctr.gsk.co.uk/Summary/fluticasone_salmeterol/studylist.asp Identifier: No. SCO100540.

Effect of intramuscular injection of hepatocyte growth factor plasmid DNA with electroporation on bleomycin-induced lung fibrosis in rats.

BACKGROUND: So far, there is no efficient treatment for pulmonary fibrosis. The objective of this study was to determine whether intramuscular injection of the hepatocyte growth factor (HGF) plasmid DNA by in vivo electroporation could prevent bleomycin-induced pulmonary fibrosis in rats, and to investigate the possible mechanisms. METHODS: Twenty male Wistar rats were randomly divided into four groups: control group (group C), model group (group M), early intervention group (group I) and late intervention group (group II). Groups M, I and II were intratracheally infused with bleomycin, then injected the plasmid pcDNA3.1-hHGF to group I on day 7, 14 and 21. Group II received the same treatment like Group I on day 14 and 21. All the rats were killed on day 28 after bleomycin injection. We detected Homo HGF expression in the rats with ELISA method and estimated the pathological fibrosis score of lung tissue using hematoxylin eosin (HE) and Massion staining. The mRNA expression of transforming growth factor-beta1 (TGF-beta1), cycloxygenase-2 (COX-2), and rat HGF in rat pulmonary parenchyma were evaluated by RT-PCR. Immunohistochemistry and Western blotting were performed to determine the protein expression of transforming TGF-beta1 and COX-2 in lung parenchyma. RESULTS: The plasmid pcDNA3.1-hHGF could express hHGF in NIH3T3 cells and the hHGF protein is secreted into the culture medium. The expression of hHGF protein could be monitored in quadriceps muscle, plasma and lung in Groups I and II. Pulmonary fibrosis levels of Groups I and II were obviously lower than that of group M (P < 0.05). Expression of TGF-beta1 protein and mRNA in lung tissue was markedly decreased in Groups I and II compared with Group M (P < 0.05). The level of expression of HGF and COX-2 mRNA was higher in Groups I and II than in Group M (P < 0.05). CONCLUSIONS: Injection of the plasmid pcDNA3.1-hHGF into skeletal muscle with electroporation has a potential role in the treatment of bleomycin-induced lung fibrosis. Exogenous HGF may inhibit the expression of TGF-beta1 and regulate the crosstalk between AECs and mesenchymal fibroblasts.

[Losartan inhibited angiotensin induced human lung fibroblast transformation and collagen synthesis].

OBJECTIVE: To observe the induction of cell transdifferentiation, connective tissue growth factor (CTGF) expression and collagen production of human lung fibroblast (HLF) by angiotensin II (Ang II), and to investigate the inhibitory effect of Ang II receptor antagonist-losartan on this process. METHODS: HLF cells were cultured and divided into a control group, an Ang II treated group, an Ang II + losartan co-incubated group and a losartan group. The marker of myofibroblast-alpha-smooth muscle actin (alpha-SMA) was detected by immunofluorescence and Western blot respectively, and the expression of CTGF mRNA and protein level were measured by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry respectively. Groups of Ang II and Ang II + losartan incubated with CTGF phosphothioate antisense, sense and random oligonucleotides transfected HLFs respectively, and collagen I (Col) I mRNA and alpha-SMA expression level were compared. The amount of hydroxyproline in the cell culture was measured by colorimetric assay. RESULTS: In the Ang II group, CTGF mRNA level (0.82 +/- 0.07) was significantly higher than the control group (0.29 +/- 0.05), the Ang II + Losartan group (0.51 +/- 0.04) and the Losartan group (0.26 +/- 0.04). The CTGF protein level in the Ang II group (0.24 +/- 0.05) was increased as compared to the other groups. In the Ang II group, Col I mRNA (1.03 +/- 0.12) and amount of hydroxyproline (0.62 +/- 0.01 ng/ml) were significantly different as compared with the other three groups. The alpha-SMA expression level (1.14 +/- 0.15) and Col I mRNA (0.30 +/- 0.04) of HLF cells transfected with antisense oligonucleotide incubated with Ang II decreased significantly as compared to those of sense and random oligonucleotide transfected cells. Alpha-SMA expression level (0.85 +/- 0.09) and Col I mRNA (0.20 +/- 0.02) of Losartan + Ang II co-incubated antisense oligonucleotide transfected HLF cells decreased significantly as compared to Ang II treated alone. CONCLUSION: Ang II promoted HLF cell transdifferentiation into myofibroblasts and increased collagen production through induction of CTGF expression. Blocking CTGF expression decreased Ang II induced transdifferentiation. Losartan blocked Ang II induced HLF cell transdifferentiation and collagen production.

[Liposarcoma of the pleura: clinical characteristics].

OBJECTIVE: Primary liposarcoma of the pleura is extremely rare. The first case report in China was described. METHODS: The clinical data of a case of primary liposarcoma of the pleura diagnosed in this hospital were reported, and the pertinent literature was reviewed. RESULTS: A total of 17 cases of primary liposarcomas of the pleura, including the case reported herein, had been described in international literature. Primary pleural liposarcomas occur predominantly in males and older patients, and the myxoid histological subtype is common. The most common symptoms are chest pain and shortness of breath. Radiographic and surgical evaluation are important for its diagnosis. Surgical resection with adjuvant radiation therapy is recommended for the disease. CONCLUSION: More data are required for the treatment and prognostic evaluation of the primary liposarcoma of the pleura.

Protective effect of tanshinone II A on lipopolysaccharide-induced lung injury in rats.

OBJECTIVE: To explore the protective effect of tanshinone II A on lipopolysaccharide (LPS)-induced lung injury in rats, and possible mechanism. METHODS: LPS (O(111): B4) was used to produce a rat model of acute lung injury. Sprague-Dawley rats were randomly divided into 3 groups (8 in each group): the control group, the model group (ALI group), and the tanshinone II A treatment group. Expression of adhesion molecule CD18 on the surface of polymorphonuclear neutrophil (PMNCD18) in venous white blood cells (WBC), and changes in coagulation-anticoagulant indexes were measured 6 h after injection of LPS or normal saline. Changes in malondialdehyde (MDA) content, wet and dry weight (W/D) ratio and morphometry of pulmonary tissue as well as PMN sequestration in the lung were also measured. RESULTS: (1) When compared with the control group, expression of PMNCD18 and MDA content were enhanced in the ALI group with a hypercoagulable state (all P<0.01) and an increased W/D ratio (P<0.05). Histopathological morphometry in the lung tissue showed higher PMN sequestration, wider alveolar septa; and lower alveolar volume density (V(V)) and alveolar surface density (S(V)), showing significant difference (P<0.01). (2) When compared with the ALI group, the expression of PMN-CD18, MDA content, and W/D ratio were all lower in Tanshinone II A treatment group (P<0.05) with ameliorated coagulation abnormality (P<0.01). Histopathological morphometry in the lung tissue showed a decrease in the PMN sequestration and the width of alveolar septa (both P<0.01), and an increase in the V(V) and S(V) (P<0.05, P<0.01). CONCLUSION: Tan II A plays a protective role in LPS-induced lung injury in rats through improving hypercoagulating state, decreasing PMN-CD18 expression and alleviating migration, reducing lipid peroxidation and alleviating pathological changes.

[Inhibitory effects of Salvia miltiorrhiza injection coordinated with dexamethasone on interleukin-13 and eotaxin expression in lung of asthmatic rats].

OBJECTIVE: To investigate the molecular mechanism of inhibitory effect of Salvia miltiorrhiza Injection (SMI) coordinated with dexamethasone (DXM) on allergic airway inflammation in asthmatic rats. METHODS: Forty SD rats were randomly divided into 5 groups equally: the normal group, the asthma model group, the DXM group, the SMI group and the DXM + SMI group, they were treated with correspondant herbal medicines. Pathologic changes of lung tissue were obseved with HE stain, count of WBC and eosinophil (Eos) in bronchoalveolar lavage fluid (BALF) were estimated and the expressions of interleukin-13 (IL-13) and Eotaxin in lung tissue were measured by RT-PCR and SP method of immunohistochemistry assay. RESULTS: There was moderate inflammation in lung tissue in the SMI group, and mild inflammation in the DXM + SMI and the DXM group, which was similar to that in the normal group. Compared with the asthma model group, Eos and WBC count in BALF and the expression of IL-13 and Eotaxin in the lung tissue were significantly lower in the three treated groups (P < 0.05), particularly in the DXM + SMI group, showing a significant difference as compared with the other two groups (P < 0.05 or P < 0.01). Additionally, IL-13 expression was positively correlated with Eotaxin expression (r = 0.92, P < 0.01). CONCLUSION: SMI could inhibit the expression of IL-13 and Eotaxin in the lung of asthmatic rats, showing inhibitory effects synergistic with DXM on airway inflammation.

[Tumor necrosis factor alpha-induced expression and activity of alveolar macrophages matrix metalloproteinase-9 in chronic obstructive pulmonary disease].

OBJECTIVE: To explore the expression and activity of matrix metalloproteinase-9 (MMP-9) from alveolar macrophages (AMs) induced by tumor necrosis factor-alpha (TNFalpha) in patients with chronic obstructive pulmonary disease (COPD), and therefore to study the possible mechanisms. METHODS: AMs were collected from bronchoalveolar lavage fluid from patients with COPD, and then were stimulated for 24 h with TNFalpha at concentrations of 0.1 ng/ml, 1 ng/ml, 5 ng/ml, 10 ng/ml, and 20 ng/ml, respectively. The gene expression level of MMP-9 induced by TNFalpha was detected by semi-quantitative RT-PCR, and MMP-9 activity was measured by zymography. NF-kappaB activity was detected by electrophoretic mobility shift assay. RESULTS: In unstimulated AMs, the densitometry unit rate of MMP-9 mRNA to beta-actin mRNA was 0.1826 +/- 0.0084. When AMs were stimulated by TNFalpha at concentrations of 0.1 ng/ml, 1 ng/ml, 5 ng/ml, 10 ng/ml, and 20 ng/ml, the densitometry unit rates of MMP-9 mRNA to beta-actin mRNA were 0.4279 +/- 0.0204, 0.5431 +/- 0.0058, 0.6185 +/- 0.0208, 0.6859 +/- 0.0080, and 0.7784 +/- 0.0208, respectively. In unstimulated AMs, the densitometry unit of active MMP-9 was 10 227 +/- 738, but when AMs were stimulated by TNFalpha at concentrations of 0.1 ng/ml, 1 ng/ml, 5 ng/ml, 10 ng/ml, and 20 ng/ml, the densitometry units of active MMP-9 were 40,528 +/- 974, 113,741 +/- 4547, 190,696 +/- 4032, 309,505 +/- 7639, and 427,305 +/- 9145, respectively. NF-kappaB activity was significantly elevated after AMs were stimulated by TNFalpha at the concentration of 10 ng/ml (P < 0.05). CONCLUSIONS: The expression and activity of MMP-9 can be induced by TNFalpha, and the induction was possibly related with the NF-kappaB-mediated signal transduction pathway.

[Effects of Shen-Mai injection on sternohyoid contractile properties in chronic intermittent hypoxia rat].

OBJECTIVE: To investigate the effect of Shen-Mai injection (SMI) on sternohyoid contractile properties in rats with chronic intermittent hypoxia. METHODS: Thirty healthy male SD rats were randomly assigned to three equal groups, the control group (A group), the chronic intermittent hypoxia group (B group) and the SMI group(C group). Rats in B group and C group were exposed to alternating periods of hypoxia and normoxia once per minute for 8 h/d for 5 weeks in order to mimic the intermittent hypoxia of obstructive sleep apnea-hypopnea syndrome (OSAHS) in humans. Isometric contractile properties were determined by electrostimulating the strips of isolated sternohyoid muscles at different frequencies (from 10 Hz to 100 Hz) to observe the changes of the sternohyoid contractile properties. RESULTS: (1) The tension of sternohyoid muscle in A group at different frequencies was (23.2 +/- 5.6), (26.2 +/- 5.0), (35.1 +/- 5.4), (46.0 +/- 8.5), (57.0 +/- 9.9), (69.9 +/- 9.7), (79.2 +/- 9.5), (85.7 +/- 7.6), (87.9 +/- 7.9), and (86.6 +/- 12.4) g/cm(2). The tension of sternohyoid muscle in B group [(19.5 +/- 4.7), (23.8 +/- 4.7), (33.0 +/- 5.1), (45.1 +/- 5.9), (54.2 +/- 7.0), (66.1 +/- 9.1), (74.2 +/- 9.1), (79.7 +/- 9.0), (82.0 +/- 8.4), and (80.7 +/- 11.8) g/cm(2)] was not significantly different from those in A group respectively (all P > 0.05); while the tension of sternohyoid muscle in C group [(30.5 +/- 2.3), (40.0 +/- 5.4), (56.2 +/- 7.6), (72.2 +/- 6.4), (82.0 +/- 5.5), (92.4 +/- 4.6), (98.1 +/- 4.0), (99.2 +/- 7.4), (101.8 +/- 3.9), and (102.2 +/- 4.0) g/cm(2)] was significantly different from those in B group respectively (all P < 0.05). (2) In fatigue test, the tension percentages of sternohyoid muscle in A group at 1, 2, 3, 4, and 5 min were (87.9 +/- 5.7)%, (72.1 +/- 11.5)%, (55.6 +/- 9.6)%, (39.7 +/- 10.7)%, (33.2 +/- 10.2)%. Compared with A group, the tension percentages of sternohyoid muscle in B group at 1, 2, 3, 4, and 5 min [(75.6 +/- 8.5)%, (41.6 +/- 7.3)%, (29.0 +/- 2.7)%, (20.4 +/- 2.9)%, (18.5 +/- 2.5)%, respectively] decreased significantly (all P < 0.05). Compared with B group, the tension percentages of sternohyoid muscle in C group [(87.9 +/- 4.4)%, (67.9 +/- 14.1)%, (48.4 +/- 9.9)%, (38.2 +/- 7.0)%, (33.8 +/- 9.3)%, respectively] increased significantly (all P < 0.05). CONCLUSIONS: Chronic intermittent hypoxia can increase upper airway muscle fatigue. SMI can significantly increase the contractile properties of upper airway muscle and resist the fatigue of upper airway muscle.

[The effect of inhaled glucocorticosteroid on protein kinase C alpha expression and interleukin-5 production in induced sputum inflammatory cells of asthma patients].

OBJECTIVE: To investigate the expression of protein kinase C alpha (PKC alpha) in inflammatory cells and the level of interleukin-5 (IL-5) in induced sputum in asthma patients and the effect of inhaled glucocorticosteroid on them. METHODS: 29 asthma patients were classified into 2 groups; 14 patients were treated with fluticasone propionate for 2 weeks and the others were treated with fluticasone propionate for 4 weeks. Induced sputum with inhaled hypertonic saline (4% - 5%) was collected. 13 healthy volunteers were included as controls. Lung ventilatory function and forced expiratory volume in one second (FEV(1)) were measured in all patients. The expression of PKC alpha in the inflammatory cells was detected using immunocytochemistry and the positive percentage in different cells was counted. IL-5 in sputum supernatants was detected using enzyme-linked immunosorbent assay. RESULTS: The percentage of eosinophils (11.1 +/- 4.3)%, lymphocytes (4.3 +/- 2.6)%, PKC alpha positive cells (79.0 +/- 9.6)% and the concentration of IL-5 (64.9 +/- 46.0) ng/L in asthmatics were higher than the percentage of eosinophils (0.7 +/- 0.5)%, lymphocytes (1.1 +/- 0.5)%, PKC alpha positive cells (10.5 +/- 2.3)% and the concentration of IL-5 (14.3 +/- 8.4) ng/L in the controls (P < 0.01). The percentage of PKC alpha positive cells and the concentration of IL-5 in the two groups of asthma were lower than that before fluticasone propionate treatment (P < 0.05), and the forced expiratory volume in first second was correlated to them (respectively, r = -0.423, P < 0.05; n = 29, r = -0.664, P < 0.01). The concentration of IL-5 was correlated to the percentage of PKC alpha positive cells (n = 29, r = 0.623, P < 0.01). CONCLUSIONS: PKC alpha in the inflammatory cells and IL-5 may play an important part in airway inflammation, and the signal transduction of the PKC alpha may be one of the mechanisms of the airway inflammation in asthma. Glucocorticosteroid could suppress the expression of PKC alpha in airway inflammatory cells and then decrease the production of IL-5 in asthmatic airways.

[The expression of nerve growth factor in inflammatory cells of induced sputum from patients with asthma].

OBJECTIVE: To investigate the expression of nerve growth factor (NGF) in inflammatory cells of induced sputum from patients with asthma. METHODS: Induced sputum samples were collected from 11 asthmatics in exacerbation, 19 asthmatics with stable disease and 11 healthy individuals. Sputum samples of 12 asthmatics with stable disease were collected again after treatment with inhaled corticosteroids (ICS) for 2 weeks. The differential counts of sputum inflammatory cells were studied. NGF was detected using immunocytochemistry and the positive percentages in different cells were measured. Interleukin-5 (IL-5) in sputum supernatants was detected using enzyme-linked immunosorbent assay. RESULTS: NGF was localized in the plasma of macrophages, lymphocytes and granulocytes. 1. In asthmatics with both exacerbation and stable disease, the NGF positive percentages in macrophages [(77.8 +/- 4.8)% and (68.1 +/- 5.3)%], lymphocytes [(43.2 +/- 9.3)% and (34.2 +/- 11.3)%], and granulocytes [(38.7 +/- 6.2)% and (32.1 +/- 3.7)%] were all significantly higher (P < 0.01) than those in the healthy controls [(37.6 +/- 8.4)%, (12.6 +/- 8.5)%, and (12.7 +/- 8.2)%, respectively]. The differences of those indices between patients with exacerbation and patients with stable disease were also significant (P < 0.01). Patients with exacerbation had significantly elevated levels of IL-5 [(124 +/- 71) ng/L] compared with patients with stable disease [(52 +/- 23) ng/L] and the healthy controls [(26 +/- 12) ng/L] (P < 0.01). 2. In the 12 patients with stable disease evaluated after treatment with ICS, the NGF positive percentages in macrophages [(54.3 +/- 7.3)%], lymphocytes [(22.0 +/- 5.6)%], and granulocytes [(27.0 +/- 4.3)%], and the level of IL-5 [(29 +/- 13) ng/L] were all decreased significantly (P < 0.01) compared with the baselines [(67.0 +/- 5.0)%, (36.0 +/- 9.0)%, (32.0 +/- 3.0)%, and (49 +/- 26) ng/L, respectively]. 3. The NGF positive percentages in both macrophages and granulocytes correlated to the sputum lymphocyte counts (r = 0.723, r = 0.630, P < 0.01; respectively) and the levels of IL-5 (r = 0.652, r = 0.636, P < 0.01; respectively). CONCLUSION: The expression of NGF in macrophages and lymphocytes in the airways is upregulated in asthma, which indicates a potential correlation between NGF and airway inflammation in asthma.

[Effect of Shen-Mai injection and aminophylline on diaphragmatic muscle cell apoptosis and related gene expression in rats with chronic hypoxia].

OBJECTIVE: To investigate the effect of Shen-Mai injection (SMI) and aminophylline on diaphragmatic muscle cell apoptosis and the Fas/FasL expression in chronic hypoxic rats. METHODS: Seventy-five male Wistar rats were randomly divided into three equal groups, control group (A group), SMI group (B group) and aminophylline group (C group). Then each group was further divided into five subgroups of pre-hypoxia, hypoxia 1 w, 2 w, 3 w and 4 w groups (5 rats each). The concentration of oxygen was (10 +/- 3)%, 7 d/w, 8 h/d for all groups, but only B group and C group received SMI (2 ml/d) and aminophylline (10 mg/kg) respectively. Apoptosis and Fas/FasL expression of diaphragmatic muscle cells were examined by the streptavidin biotin-peroxidase complex (SABC) immunohistochemistry techniques and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay, and Dunnett-t test was employed to compare the effects of SMI and aminophylline. RESULTS: (1) Fas, FasL expression in normal diaphragmatic muscle cells was very low with a positive rate of (2.77 +/- 0.45)% and (2.32 +/- 0.61)%. After hypoxia, the positive rates increased with the time of hypoxia time. SMI showed an inhibition on diaphragmatic muscle cell Fas and FasL expression;after hypoxia 1 w, 2 w, 3 w and 4 w, Fas expression [(6.36 +/- 4.17)%, (9.77 +/- 4.12)%, (18.02 +/- 6.91)% and (21.09 +/- 8.09)%] and FasL expression [(5.32 +/- 6.16)%, (9.58 +/- 3.79)%, (12.01 +/- 8.71)%, (19.43 +/- 10.31)%] in B group were different from those in A group respectively (all P < 0.05). But aminophylline did not show such an effect, the expression of Fas [(10.87 +/- 3.62)%, (24.13 +/- 3.79)%, (35.39 +/- 9.02)%, (39.56 +/- 10.12)%] and FasL [(9.37 +/- 4.07)%, (20.16 +/- 4.88)%, (31.81 +/- 7.07)%, (35.51 +/- 9.13)%] were not significantly different from those in A group respectively (all P > 0.05). (2) Diaphragmatic muscle cell apoptosis was very low in normal rats with a rate of (0.93 +/- 0.29)%, which also increased after hypoxia and the increase was associated with the time of hypoxia. Apoptosis rate was decreased by the administration of SMI, the rates of B group were (5.01 +/- 3.71)%, (9.37 +/- 3.12)%, (14.66 +/- 8.76)%, (18.16 +/- 7.02)%, respectively. Except for the first week, the differences of other weeks were all statistically significant when compared with A groups (all P < 0.05). But the effect of aminophylline was different, as compared to A group, only the apoptosis rate in hypoxia 4 w [(30.92 +/- 11.13)%] of C group being statistically significant different (P < 0.05). CONCLUSIONS: Fas and FasL participated in diaphragmatic muscle cell apoptosis in rats with chronic hypoxia. SMI showed a definite effect on the Fas and FasL protein expression and decreased diaphragmatic muscle cell apoptosis, which contributed to the therapeutic effect on diaphragmatic fatigue caused by hypoxia.

[The effect of nitric oxide on potassium channels of bronchial smooth muscle cells from asthmatic rats].

OBJECTIVE: To investigate the effect of nitric oxide (NO) on resting membrane potential (Em) and potassium currents of bronchial smooth muscle cells (BSMC) from asthmatic rat models. METHODS: Sixteen male SD rats were randomly divided into two groups: a control group and an asthmatic model group. Single BSMC was obtained by acute enzyme-digestion separation method. All experiments were conducted in conventional whole-cell configuration of patch clamp technique. Em currents of Ca2+ activated potassium channels (BKCa) and voltage-dependent potassium channel (Kv) of two groups were separately measured, and the changes of The resting Em and potassium currents of BSMC before and after addition of NO donor sodium nitroprusside (SNP) were also measured. RESULTS: (1) The Em of the asthmatic model group [(-29 +/- 6) mV, n = 12] was significantly lower than that of the control group [(-35 +/- 6) mV, n = 15, P < 0.05]; SNP significantly increased Em of the asthmatic model group to [(-38 +/- 7) mV, n = 12, (P < 0.0 1)], there is no significant difference of Em between normal group and that of asthmatic model group after SNP treatment (P > 0.05), which meant that SNP could repolarize BSMC to normal. (2) The mean current density of BKCa from asthmatic model group [(44 +/- 17) pA/pF, n = 8] under pulse protocol was significantly lower than that of the control group [(73 +/- 20) pA/pF, n = 10, P < 0.01], and SNP significantly increased the mean current density of the asthmatic model group to [(79 +/- 16) pA/pF, n = 10, P < 0.01], which was close to control group (P > 0.05); under ramp protocol, the current densities of control and asthmatic model group were [(75 +/- 19) pA/pF, n = 10] and [(46 +/- 16) pA/pF, n = 8] respectively, there was significant difference between two groups (P < 0.01), SNP treatment significantly increased current density of asthmatic model group to [(82 +/- 21) pA/pF, n = 8, P < 0.01]. (3) The mean current density of Kv of the asthmatic model group [(32 +/- 9) pA/pF, n = 8] under pulse protocol was significantly lower than that of the control group [(58 +/- 10) pA/pF, n = 8, P < 0.05], and SNP significantly increased the mean current density of Kv of the asthmatic model group to [(45 +/- 13) pA/pF, n = 8, P < 0.05]. Under ramp protocol, current density of Kv of asthmatic model group was [(38 +/- 11) pA/pF, n = 8], which was significantly lower than that of control group [(62 +/- 14) pA/pF, n = 8, P < 0.05], SNP treatment significantly increased Kv current density of asthmatic model group to [(53 +/- 9) pA/pF, n = 8, P < 0.05]. there was a similar change. CONCLUSIONS: SNP can improve the resting membrane potential of BSMC from rat asthmatic models and increase the potassium channel activity of BSMC, which is impaired in asthma status. The result suggests that potassium channel mediates the relaxation effect of NO on asthmatic airway smooth muscle.

[Effect of shenmai injection on L-type calcium channel of diaphragmatic muscle cells in rats].

OBJECTIVE: To explore the effect of Shenmai Injection (SMI) on L-type calcium channel of diaphragmatic muscle cells in rats. METHODS: Single diaphragmatic muscle cell of rats was obtained by the acute enzyme isolation method and the standard whole-cell patch clamp technique was used to record the inward peak L-type calcium current (IPLC) and current-voltage relationship curve of diaphragmatic muscle cells of 7 rats, and to compare the effects of SMI in various concentrations on them. RESULTS: When keeping the electric potential at -80 mV, stimulation frequency 0.5 Hz, clamp time 300 ms, stepped voltage 10 mV, and depolarized to +60 mV, 10 microliters/ml of SMI could only cause the mean IPLC of rat's diaphragmatic muscle cells increased from -6.9 +/- 0.6 pA/pF to -7.5 +/- 0.7 pA/pF, the amplification being (9.2 +/- 2.8)%, comparison between those of pre-treatment and post-treatment showed insignificant difference. But when the concentration of SMI increased to 50 microliters/ml and 100 microliters/ml, the mean IPLC increased to -8.4 +/- 0.6 pA/pF and -9.2 +/- 0.6 pA/pF, respectively, and the amplification was (22.4 +/- 1.7)% and (34.6 +/- 4.6)% respectively, showing significant difference to that of pre-treatment (P < 0.05). However, SMI showed no significant effect on maximal activation potential and reversal potential. CONCLUSION: SMI can activate the calcium channel of diaphragmatic muscle cells in rats, increase the influx of Ca2+, so as to strengthen the contraction of diaphragmatic muscle, which may be one of the ionic channel mechanisms of SMI in treating diaphragmatic muscle fatigue in clinical practice.