Respiratory virus poses a serious threat to human life and health. Airway epithelial cells are the body's first line of defense from a wide variety of foreign pathogens, such as viruses and bacteria. Therefore, successful airway epithelial cell culture can provide a model for investigating the mechanisms underlying respiratory pathogenic diseases following airway virus infection. This respiratory disease model can also be used for the potential development of novel therapeutics. Here we provide a brief review of recent developments on the culture of cells derived from human trachea-bronchial airway epithelium, and the application of this model for studying respiratory virus and disease.
Animals ; Cell Culture Techniques ; Epithelial Cells ; virology ; Humans ; Respiratory Tract Diseases ; virology ; Virus Diseases ; virology ; Virus Physiological Phenomena ; Viruses ; genetics ; isolation & purification

Objective To assess the risk assessment of stroke associated pneumonia (SAP ) in patients with ischemic stroke using A2DS2 score.Methods A total of 1279 patients with ischemic stroke who were admitted and treated in our department from 2009 to 201 1 were retrospectively analyzed and applicated A2DS2 score.A2DS2 score was calculated as follows:Age ≥75 years=1,atrial fibrillation=1, dysphagia=2,male sex=1;stroke severity:NIHSS score 0-4=0,5-15 =3,≥16=5.The patients were divided into three groups according to A2DS2 score:620 in score 0 group,383 in score 1-9 group,276 in score ≥10 group.The three groups were comparatively analyzed.The diagnostic criteria for SAP were as follows:newly emerging lesions or progressively infiltrating lesions in post-stroke chest images combined with more than two of the following clinical symptoms of infection:(1 )fever ≥38 ℃;(2 )newly occurred cough,productive cough or exacerbation of preexisting respiratory tract symptoms with or without chest pain;(3)signs of pulmonary consolidation and/or moist rales;(4)peripheral white blood cell count≥10 ×109 L -1 or≤4 ×109 L-1 with or without nuclear shift to left,while excluding some diseases with similar clinical manifestations to pneumonia, such as tuberculosis, pulmonary tumors, non-infectious interstitial lung disease,pulmonary edema,pulmonary embolism and atelectasis.Analysis of the incidence and mortality of SAP as well as the correlation with ischemic stroke site were performed in the three groups,respectively. Mean ± standard deviation (x ±s)was used to represent measurement data with normal distribution and t test was used.Percentage was used to represent enumeration data and χ2 test was used.Results The incidence of SAP was significantly higher in A2DS2 score ≥10 group compared with those in score 1-9 and score 0 groups (7 1.7%vs..22.7%,7 1.7%vs..3.7%,respectively),whereas the mortality in score ≥10 group was significantly higher than those in score 1-9 and score 0 groups (16.7%vs.4.96%,16.7%vs.0.3%,respectively).The incidences of cerebral infarction in posterior circulation and cross-MCA, ACA distribution areas were significantly higher in SAP group compared with those in non-SAP group (35.1%vs.10.1%,11.4%vs.7.5%,respectively).The incidence of non-fermentative bacteria infection was significantly increased in score ≥10 group.Conclusions A2DS2 score provides a basis for risk stratification of SAP.The prevention of SAP needs to be strengthened in acute ischemic stroke patients having a A2DS2 score ≥10.

Objective To investigate the drug resistance status quo of imipenem-resistant Pseudomonas aeruginosa and carbap-enemase gene carrying in Xi′an area for guiding the clinical rational use of antibacterial drugs.Methods 151 strains of imipenem-re-sistant Pseudomonas aeruginosa isolated from clinical samples in 4 hospitals from August 2012 to July 2013 were continuously col-lected.Then the drug resistance characteristics of imipenem-resistant Pseudomonas aeruginosa were investigated by the antimicrobi-al drug sensitivity test.The PCR technique was adopted to detect the carrying situation of carbapenemase drug-resistance genes in imipenem-resistant Pseudomonas aeruginosa.Results Totally isolated 151 strains of imipenem-resistant Pseudomonas aeruginosa were mainly distributed in the neurosurgery ICU (37.1%),neurology ICU (27.1%)and the burn department (19.9%);the detec-ted strains were sensitive to polymyxin B and resistant to other 9 kinds of antibacterial drugs in different degrees;94 strains carried VIM gene,32 strains carried IMP gene,5 strains carried SPM gene and 3 strains carried SIM gene.Conclusion The multidrug re-sistance and pan-drug resistance phenomenon of imipenem-resistant Pseudomonas aeruginosa is serious,its cause might be related with the carbapenemases-producing drug resistant gene expression,the drug resistance genes are dominated by VIM and IMP.Clinic should strengthen the bacterial drug resistance monitoring and use antibacterial drugs rationally and effectively for preventing the spreading of imipenem-resistant Pseudomonas aeruginosa.

Objective To investigate the distribution of characteristics,clinical manifestations,laboratory parameters and activity index of patients with systemic lupus erythematosus (SLE) and to determine their role in helping to make correct clinical diagnosis and disease the activity evaluation of SLE.Methods Collect the data of 1037 SLE patients of Ningxia Medical University Affiliated Hospital from January 2006 to June 2010.Data were analyzed with t test and Chi-square test.Results Over the past three years,there were more and more patients were admitted year by year.Among the 1037 cases of SLE patients,most of them 20-40 year-old woman,accounting for 67.5％ of the whole patient population,with a male to female ratio was 1:8.26.Joint pain was the most common initial symptom,accounting for 54.3％,followed by skin rash,accounting for 48.2％.Decreased complement C3 level and platelets counts, proteinuria,and positive anti-dsDNA antibody could be used as indicators for early diagnosis of SLE.SLEDAI activity score higher than 9 were presented in 26.0％ of patients.Factors that could impact the final score of SLEADI were fever,arthritis,skin rash,proteinuria,low complement levels,high titers of anti-dsDNA antibody,pleurisy,alopecia,mucosal ulcers,pericarditis,mental illness and decreased platelets count.Patients with active disease had a higher accidence of fever,arthritis,skin rash,lung damage,alopecia,mucosal ulcers,heart damage,mental illness and renal damaged,low complement levels,high level of anti-dsDNA antibody titers and elevated erythrocyte sedimentation rate.Conclusion SLE is a multi-system disease with multiple organ involvement,with characteristic clinical symptoms and immunological abnormalities,thus early diagnosis is very important.Understanding the characteristics of the diseases,correct judgement of the disease activity,reasonable and effective treatment all can delay the development of organ damage and improve the prognosis.

Objective To investigate the bactericidal mechanism of electrolyzed oxidizing water (EOW) against Pseudomona aeruginosa (P.aeruginosa).Methods Bactericidal mechanism of EOW against P.aeruginosa was studied through intracellular protein leakage,nucleic acid,and cell membrane calcium ion permeability,2 ％ glutaraldehyde was used as positive control group,and normal saline (NS) was used as negative control group.Results The killing rates of EOW and 2％ glutaraldehyde to P.aeruginosa were both＞99.99％ with 30-second contact time,and 100.00％ with 60-second contact time.After 60-second contact with EOW,NS,and 2％ glutaraldehyde,the protein leakage of P.aeruginosa detected by bicinchoninic acid (BCA) were (96.00 ± 7.42),(94.15 ± 7.49),and (216.97 ± 10.35)μg/mL,respectively,difference was significant(F =613.20,P＜0.01),2％ glutaraldehyde group was higher than EOW group and NS group;protein leakage did not change with the increase of contact time(all P＞0.05).Electrophoretogram of random amplified polymorphic DNA showed high intensity dense band between 500-1000 Kb in EOW group and NS group,while 2％ glutaraldehyde group was without amplified bands.The fluorescence intensity of calcium ion of EOW group and 2％ glutaraldehyde group were both lower than that of NS group.Conclusion Bactericidal mechanism of EOW may be due to the damage of membrane permeability of P.aeruginosa,which causes Ca2+ leakage,but fails to cause protein leakage,the damage to nucleic acid is not obvious,DNA may not be a bactericidal target of EOW.

Objective To establish animal models in order to provide an experimental study basis for both the pathogenesis study and taking effective prevention scheme for doxorubicin extravasation injury. Methods A total of 20 Kunming mice for experiments on doxorubicin extravasation injury were divided in-to four groups, I.e., high dose group(2 g/L), medium dose group(1 g/L), low dose group(0.5 g/L) and the control group (injection with water). Dosages were administered with subcutaneous injection on both sides of mice abdomen. The adverse reaction of body, damage areas of extravasation injury, recovery period were observed and histopathologic slides of animal models on both 4 days and 11 days after experiment were performed and compared. Results No significant adverse body reaction was observed after injection for all groups. The damage areas due to extravasation injury were dosage and concentration dependent. In addi-tion, significant differences in recovery period were observed for mice in different groups, that is, the higher injection concentration and dose led to the longer recovery period. Results from the histopathologic study in-dicated that the putrescence of damage area was developed in high dose group mice, and the ulcer occurred after 4 d of dosage in medium dose group mice, respectively. In contrast, no ulcer was observed in low dose group mice. Conclusions It would be feasible to establish a prevention model for mice on doxorubicin extravasation injury by subcutaneous injection at a dosage of 0.05ml(1 g/L).

Pigs are increasingly recognized as "natural" hosts of infection by human respiratory viruses because of their similarities to humans in terms of lung physiology, airway morphology, cell types, and distribution of cell receptors in the respiratory tract. We wished to explore the mechanisms of infection by respiratory viruses and screening of drug that could be used to treat respiratory-system diseases. Hence, we developed a model of well-differentiated porcine airway epithelial cells (PAECs) derived from pig-lung tissue and cultured them with serum-free medium under an air-liquid interface condition in vitro. We identified the PAEC model using scanning electron microscopy, electrophysiology, and immunohistology. To evaluate application of gene therapy of adeno-associated virus (AAV)6 on the PAEC model, we generated recombinant adeno-associated virus 6-green fluorescent protein (rAAV6-GFP) using the three-plasmid transfection method and infected PAECs from the apical surface with rAAV6-GFP. Results demonstrated that the PAEC model comprised a multilayer epithelial structure containing ciliated mucous secretory cells, with basal cells located directly beneath the multilayer. rAAV6-GFP could infect PAECs from the apical surface and efficiently transduce PAECs to mediate the long-term expression of the exogenous gene. Establishment of a model of well-differentiated PAECs in vitro could lay a solid foundation for the study of infection by respiratory pathogens, as well as the screening and gene therapy of agents used to treat diseases of the respiratory system.
Animals ; Cell Differentiation ; Dependovirus ; genetics ; Epithelial Cells ; cytology ; metabolism ; Green Fluorescent Proteins ; genetics ; HEK293 Cells ; Humans ; Lung ; cytology ; Membrane Potentials ; Mucins ; metabolism ; Swine ; Transduction, Genetic ; Tubulin ; metabolism

Objective To observe clinical curative effect of modified total cystectomy and Mainz Ⅱ neobladder. Methods Seventeen patients with bladder neoplasms were treated with modified total cystectomy and Mainz Ⅱ neobladder for urinary diversion. The paries posterior allantois with intestinum rectum and colon sigmoideum were taken 10 cm respectively, split the mesenterium edges, conduplicated and bilayer sutured from the junction of intestinum rectum and colon sigmoideum, bilateral ureters antireflux anastomosed respectively with colon sigmoideum and rectal papilla, then bilayer sutured paries anterior became Mainz allantois. Results There was no surgical mortality. The operative time was 340 ～ 420 mins (mean, 350 mins).Blood transfusion was 400 ～ 800 ml ( mean 600 ml). The follow-up was 4 ～ 18 months, urine and dejecta were shunt, uresis continence was fine and the operation had fewer severe complications. Conclusion Modified total cystectomy and Mainz Ⅱ neobladder to be an effective method for urinary diversion because of its simple operation, fewer severe complications, good uresis continence and high quality of life.

Objective To build a bio-material artificial chest wall and discuss its feasibility and efficiency in reconstruction of huge bony defects of chest wall in mongrels in comparison with traditional "sandwich" procedure. Methods (1) The procine osteal and membranate tissues were treated with epoxy cross linking method and their surfaces were modified with amino acid solutions at various concen-trations and at different temperatures to obtain an artificial pleura and artificial ribs for construction of the artificial chest wall. (2) The huge bony defects (5 cm×5 cm) were created in chest wall of five Chinese mongrels. (3) Three mongrels in test group was repaired with artificial chest wall, while two mongrels in control group was repaired with traditional "sandwich" complex. A follow-up was carried out to observe reconstruction effect and rejection in both groups at 3,6 and 12 months after implantation. Results There was no death found during the perioperative period and at 12 month follow-up in test group, with abnormal contour of chest wall and good thoracic activity after reconstruction. In the meantime, there found no rejection, collapse in the repaired region or paradoxical respiration. The postoperative X-rays at 3,6,12 and 24 months showed a good integrity of the thorax, with no collapse, deformation or abnormal movement. Meanwhile, the follow-up of control group showed a normal contour but slight collapse, with no paradoxical respiration. The Chest X-ray examination revealed that the bone cement in" sandwich" complex was X ray opaque and showed mild abnormal movement with breathing. The common blood test and immune items showed no abnormal. Conclusions The bio-material artificial chest wall is a safe and effective reconstruction technique for bony defects of thoracic wall in mongrels, with no acute or chronic rejection.

Objective To explore the value of color Doppler ultrasonography(CDU)as preoperative diagnosing and postoperative monitoring in patients with stenosis of subclavian or innominate artery(SIA)before and after percutaneous transluminal angioplasty and stent(PTAS).Methods A total of 45 patients with stenosis of SIA were selected.Their extracranial vertebral artery and subclavian artery were observed with CDU.In 36 patients with one-sided stenosis of SIA.blood flow direction and spectrum of extracranial vertebraI artery were analyzed.Subclavian steal grade confirmed by spectrum change of vertebral artery was compared with the stenosis grade on quantitative angiography(QA).Among 45 patients with stenosis of SIA,PTAS was performed successfully in 36 patients(36 stents implanted).After stents implanted, hemodynamie parameters such as peak systolic velocity(PSV)in stent, follow-up velocity ratio in stem(VR)were measured.Restenosis in stent were observed.Results In 36 patients with one-sided stenosis of SIA,the ultrasonographic steal grade correlated with the QA stenosis grade significantly(r=0.752,P<0.01).Among 36 patients with stent implanted,follow up 8-12 months, 4 patients with restenosis in stent were found.Compared with the patients without restenosis in stent,PSV and VR in stent increased significantly(P<0.01).Conclusions As a fast and non-invasive imaging technique,CDU could allow diagnosis of stenosis of SIA before PTAS,and detection of restenosis after PTAS.