Objective To assess the clinical significance of SAA, IP-10 and PCT in the diagnosis of AECOPD. Methods Sixty AECOPD patients, 52 with sCOPD, and 28 healthy subjects were assigned to three groups. Clinical data and serum specimen were obtained from another 19 AECOPD patients at stable stage as AECOPD-sCOPD group. Serum levels of SAA, IP10 and PCT were quantitatively measured by ELISA. Levels of multiple serum markers were statistically compared among different groups. Results The concentration of SAA significantly differed between the AECOPD and sCOPD groups (P < 0.05), so did the concentration of IP-10 (P < 0.05); no statistical significance was found in terms of serum PCT concentration between the two groups (P > 0.05). Conclusions As compared with the sCOPD group, levels of serum SAA and IP-10 in the AECOPD group were significantly elevated, which is helpful in the diagnosis of AECOPD with a sensitivity and specificity of 100% and 54.9%for SAA, and 96.1%and 75.0%for IP-10. However, PCT level failed to identify AECOPD from sCOPD.

Objective To find whether heNOS could be expressed in mouse lung after intratracheal administration of recombinant adenovirus vector.Methods heNOS cDNA was obtained by RT-PCR from total RNA which extracted from human HUVEC.The replication-deficient heNOS recombinant adenovirus vector was constructed with a ligation method.High titer of recombinant adenovirus was obtained by chromatographic methods.The expression of heNOS protein was determined by immunohistochemistry staining after intratracheal administration of recombinant adenovirus.Results Sequence confirmed the cloned cDNA containing the whole ORF and the heNOS cDNA was about 3731 bp.It showed 99.93% identity with that of heNOS cDNA in GenBank(163729).The biological activity of heNOS protein was examined in transfected cos7 cell line with heNOS cDNA in eukaryotic expression vector of pcDNA3.0.The replication-deficient recombinant adenovirus vector containing heNOS cDNA was constructed successfully and the purified viral titer was 2.0?1010pfu/mL.After intratracheal administration of the recombinant adenovirus,heNOS expression was found in the majority of bronchial epithelium,alveolar lining cells,endothelial cellsand smooth muscle cells of pulmonary vessels.In control group,little endogenous eNOS immunoreactivity was detected in pulmonary vessels and no eNOS immunoreactivity was shown in bronchial and alveolar epithelial cells.Conclusion The replication-deficient recombinant human endothelial nitric oxide synthase mediated by adenovirus vector constructed could be delivered into the lung tissue of mouse by intratracheal administration of recombinant adenovirus and can be expressed in lung tissue with high-efficiency.

Objective To explore the possible role of CD28 +/CD152 +:B7 eostimulators in immune pathophysiology of severe pneumonia.Methods 22 severe pneumonia peripheral blood sample were used to analyze the expression of CD3+ T cell CD28+,CD152+,CD14++ on mononuelear cell CD86+,and HLA - DR + by FACS expression.The relationship between CD28+,CTLA4,CD86+ and the HLA-DR +,and the relationship between APACHE Ⅱ Grading,CD28+,CD152+,CD86+ and HLA-DR + were analyzed.Results Compared with the control group,the expression of CD3 + T cell,CD86+ and HLA - DR + were remarkably reduced while the expression of CD28+ and CD152+ were markedly increased in patients with severe pneumonia who were hospitalized in 24 h(P＜0.05).However,T cells with positive CD8+ CD3+ and CD4+ CD3+ had no significant change between two groups(P＞0.05).For patients with severe pneumonia who survived,the APACHE Ⅱ scores were significantly reduced while the expression of CD28+,CD152+,CD86+,HLA-DR + and CD3+ + cells were significantly increased after 10 days from admission(P＜0.05).By contrast,T cells with positive CD8+ CD3+ and CD4+CD3+ had no significant change between two groups(P＞0.05).There were no relation between costimulators CD28+ and HLA - DR + (r=-0.12,P=0.54)and APACHE Ⅱ scores(r=-0.30,P=0.19) in control group.CD86+ and HLA - DR + showed positive correlation(r=0.65,P=0.00).CD86+ and APACHE Ⅱ scores had no correlation(r=-0.38,P=0.09).Conclusion The costimulators expressed abnormally in circumference blood of patients with severe pneumonia,CD86+ decreased,but CD28+,CD152+ increased.T cell of circumference blood was at the condition of "anergy".The increase of CD28+,CD86+,CD86+ and HLA - DR + during convalescence stages in patient with severe pneumonia showed that spocific immunity was advantageous for restoration in these patients.The relationship among CD86+,CTLA4 and HLA - DR + indicated that CD28+/CD152+:B7 play an role in the occurrence and development of severe pneumonia.

Objective To investigate the distribution of blaOXA-51-like genes and the clonal relation-ship among Acinetobacter baumannii strains isolated from three teaching hospitals in Guangzhou , China. Methods Fifty-two Acinetobacter baumannii isolates were genotyped by multilocus sequence typing (MLST).eBURST algorithm was performed to define clonal complexes (CCs).blaOXA-51-like genes were am-plified by using polymerase chain reaction ( PCR) and sequenced .Results MLST grouped the A.bauman-nii isolates into 5 existing sequence types (STs) and 7 new STs.STn4 carried allele G1 with a T→C muta-tion at the 3rd nucleotide site (nt3) on the gpi111 locus.STn5 carried allele A1, possessing A→C muta-tions at nt156 and nt159 on the gltA1 locus.ST195 and ST208 accounted for 69.2%of all isolates.Clonal relationship analysis showed that ST 195 and ST208 belonged to CC92.Fifty-one A.baumannii isolates car-ried OXA-66 and the rest one carried OXA-199.Conclusion A.baumannii strains that belonged to CC92 and carried OXA-66 were the predominant genotype circulating in Guangzhou , China.

Objective: To study the effect and the mechanism of acute hypoxia on Ca2+-ATPase inhibitor, cyclopiazonic acid (CPA) induced intracellular calcium cation enhancement in rat distal pulmonary venous smooth muscle cells (PVSMC) .
Methods: The PVSMC were isolated from 6 male SD rats and the cells were cultured for further experiment. Enhancing effects of CPA, acute hypoxia (4% O2) on [Ca2+]i in distal PVSMC and the interventional effects of 2 store-operated Ca2+ channels (SOCC) inhibitors, NiCl2 and SKF96365 on [Ca2+]i in distal PVSMC were tested by lfuorescence microscope and intracellular [Ca2+] examining system.
Results: When PVSMC were perfused with Ca2+-free Krebs solution containing 5 μmol/L nifedipine, 10 μmol/L CPA caused a slight elevation of [Ca2+]i, and acute hypoxia obviously enhanced the [Ca2+]i in PVSMC. When restoration of extracellular [Ca2+] to 2.5 mmol/L, 10 μmol/L CPA caused signiifcant elevation of [Ca2+]i, and acute hypoxia obviously enhanced [Ca2+]i induced by CPA in PVSMC. The SOCC inhibitors, NiCl2 (500 μmol/L) and SKF96365 (50 μmol/L) distinctively attenuated the elevation of [Ca2+]i by hypoxia and CPA. However, NiCl2 and SKF96365 had no effect on high potassium (60 mmol/L KCl Krebs solution) induced elevation of [Ca2+]i in distal PVSMC.
Conclusion: Acute hypoxia enhanced the elevation of [Ca2+]i induced by CPA; such effect could be selectively blocked by SOCC inhibitor which indicated that acute hypoxia could enhance the activity of SOCC in rat distal PVSMC.

Objective To study the effect of SKF96365 and NiCl2 on cyclopiazonic acid (CPA) induced intracellular calcium cation concentration ([Ca2+ ]i ) change in rat distal pulmonary arterial smooth muscle cells (PASMC) .Methods The rat distal PASMC were isolated and cultured .The effects of CPA ,SKF96365 and NiCl2 on [Ca2+ ]i in PASMC were tested by fluorescence microscope and InCyte [Ca2+ ]i measurement system .Results PASMC were incubated with Ca2+‐free Krebs solution containing 5μmol/L nifedipine ,10 μmol/L CPA caused a small transient increase in [Ca2+ ]i ;after restoration of extracellular Ca2+ to 2 .5 mmol/L ,10 μmol/L CPA caused marked increases in [Ca2+ ]i in PASMC incubated with Krebs solution containing 5 μmol/L nife‐dipine .Both 50 μmol/L SKF96365 and 500 μmol/L NiCl2 distinctly attenuated the increases in [Ca2+ ]i caused by 10 μmol/L CPA in PASMC .However ,neither 50 μmol/L SKF96365 nor 500 μmol/L NiCl2 affected the increases in [Ca2+ ]i caused by 60 mmol/L KCl in PASMC .Conclusion CPA induced increases in [Ca2+ ]i may related to Ca2+ release from sarcoplasmic reticulum and the in‐flux of Ca2+ through store‐operated Ca2+ channels (SOCC) in rat distal PASMC .Both SKF96365 and NiCl2 could selectively block SOCC and attenuated the influx of Ca2+ through SOCC in PASMC .

Objective To examine the value of bronchoscopy in the diagnosis of sputum-negative pulmonary tuberculosis in a general hospital.Methods A retrospective study was conducted for the 459 patients treated in a general hospital from June 2010 to May 2015.All the patients had symptoms and radiographic changes suggestive of pulmonary tuberculosis but smearnegative.All patients were subjected to bronchoscopy,including brushing,bronchoalveolar lavage (BAL) and transbronchial biopsy (TBB).The bronchoscopic specimens were submitted for direct smear and acid-fast stain,Mycobacterium tuberculosis culture or histopathological assay,respectively.Results The diagnosis was confirmed by bronchoscopy in 378 (82.4％) of the 459 patients.Of the 378 patients whose diagnosis was confirmed,pulmonary tuberculosis was identified in 238 patients (63.0％).Other diagnoses included bronchogenic carcinoma,non-specific inflammation,organizing pneumonia,pulmonary fungal infection,interstitial pneumonia,sarcoidosis,nontuberculous mycobacterial infection.Of the patients with confirmed diagnosis by bronchoscopy,the sensitivity for diagnosis was 57.95％ by direct smear and acid-fast stain of brushing or BALF,79.78％ by culture of BALF,and 56.93％ by histopathological assay of TBB specimens.The integrated method by combining smear and stain,culture and histopathological assay of TBB specimens could improve the sensitivity,specificity,positive predictive value,and negative predictive value to 91.01％,97.46％,97.98％ and 88.89％.Conclusions For the patients whose clinical manifestations and imaging changes are suggestive of pulmonary tuberculosis but smear-negative,bronchoscopy is a valuable method for the diagnosis,which should be adopted as a routine test in clinical practice.