Objective Acute exacerbations of chronic obstructive pulmonary disease (AECOPDs) are significant events which are associated with worsened quality of life and more rapid decline in lung function.And they also can increase mortality and health care costs.Pharmacologic interventions can alter the frequency of AECOPDs and COPD-related hospitalizations, mainly by focusing on three ways: antiinfective agents, antiinflammatory agents and antioxidants which also become the chief clue to the current researches for new drugs and therapies.The combination therapy of inhaled corticosteroids and long-acting β2-adrenoceptor agonists is the most widely recommended strategy for acute exacerbation prevention.But the inhaled corticosteroids possibly increase the risk for pneumonia.Further studies are needed to understand the mechanism of benefit in patient subpopulations, and to compare between efficacy of combination therapy and side-effect with inhaled corticosteroids, bronchodilators, or their combination in different patient phenotypes.We must have a better understanding of the heterogeneity implicit for COPD and the types of COPD exacerbations, and individualized treatments need to be investigated.

Objective: For cultivation of reasonable nutrition and eating habits in the new generation of singleton children in China, this nutrition intervention study was taken in a boarding primary school for 10 years to observe the outcome of their growth and development, and to find the possibility of the reasonable diet mode suitable for them. Method: This study started from 1993, and the whole diet and foods of the primary school children were designed and measured all the time when they were in school. This report was taken from 161 primary school pupils aged from 7 to 12, mostly being singleton in their family, as a sample through whole six years from Class one to six when they graduated from the school. Their growth and general health situation including body height and weight, and hemoglobin were measured and evaluated every year. Results and Conclusion: The outcome seemed positive. It suggested that the “Chinese Food Guideline” was acceptable and suitable for southern Chinese children, and the results were discussed.

The trace elements of Halibios were analysed and the effects of 5 kinds of Halibios on antiirritability, radical scavenging, memeory MAO-B activity, hemorheologic changes in animal were studied. The results showed that Halibios Acaudina molpadioides, Hippocampus Kuda ,Cavernularia habereri,Octopus valgaris ,and Octopus ocellatus contain abundant trace elements,most of them have effects on antiirritability, radical scavenging, rising memory and depressing MAO-B activity in animmal.

OBJECTIVETo compare the differences in the efficacy on speech and swallowing dysfunction of medulla oblongata palsy (MOP) after brainstem infarction between electroacupuncture at eight-neck-occiput points and routine acupoints.

METHODSSeventy-two patients were randomized into a neck-occiput points group and a meridian points group, 36 cases in each one. In the neck-occiput points group, the eight-neck-occiput points (Neck 1-4 points, Occiput 1-4 points) were selected. In the meridian points group, Lianquan (CV 23), Futu (LI 18), Tongli (HT 5), Hegu (LI 4) and the others were selected. Electroacupuncture was used in the two groups, dense-dispersion wave, retaining for 30 min. The treatment was given once a day, 5 treatments a week. Totally, 4 weeks were required. The symptom scores of speech and swallowing dysfunction were observed before and after treatment in the two groups. The efficacy was compared between the two groups.

RESULTSThe scores of speech and swallowing dysfunction were improved significantly after treatment in the two groups (P < 0.05, P < 0.01). The results in the neck-occiput points group were better than those in the meridian points group (both P < 0.01). The curative rate of speech dysfunction was 30.6% (11/36) and that of swallowing dysfunction was 22.2% (8/ 36) in the neck-occiput points group, which were better than 11.1% (4/36) and 5.6% (2/36) in the meridian points group, respectively. The differences were significant in comparison of the two groups (P < 0.01, P < 0.05).

CONCLUSIONElectroacupuncture at eight-neck-occiput points achieves a better efficacy on speech and swallowing dysfunction of MOP after brainstem infarction as compared with the routine acupoints. This therapy is characterized as more accurate point localization and safer operation.

Acupuncture Points ; Adult ; Aged ; Brain Stem Infarctions ; complications ; Bulbar Palsy, Progressive ; etiology ; physiopathology ; therapy ; Deglutition ; Electroacupuncture ; Female ; Humans ; Male ; Medulla Oblongata ; physiopathology ; Middle Aged

Objective To construct the red fluorescent protein reporter gene vector containing high mobility group box 1 protein(HMGB1) promoter sequence and study the regulation mechanism of the expression of HMGB1gene under mechanical stretch.Methods HMGB1 promoter was subcloned into a red fluorescent protein vector,pDsRed1-1.After identified by PCR,enzyme digestion and DNA sequencing,the recombinant vector pDsRed1-1-HMGB1P was then transfected into HEK293 cells.Blank vector or pDsRed-1 was transfected into 293 cells and served as controls.The expression of red fluorescent protein and its reaction to mechanical stretch were observed under a fluorescent microscope.HEK293 cells transfected with pDsRed1-1 vector served as control.Results PCR,double restriction enzyme digestion and DNA sequence analysis showed that the recombinant vector,pDsRed1-1-HMGB1P,was constructed correctly.This vector was lowly expressed in HEK293 cells of resting state.But after stimulated by mechanical stretch,strong red fluorescence was observed.No red fluorescence was observed in the control cells.Conclusion A red fluorescent protein reporter gene vector containing HMGB1 promoter sequence has been constructed successfully and expressed highly in mammalian cells.Since it responds to mechanical stretch effectively,it can thus provide a convenient tool to study the regulation mechanism of the expression of HMGB1 gene by mechanical stress.

Objective To investigate the role of extracellular regulated protein kinase (ERK) signal pathway in mechanical stretch induced high mobility group box 1 protein (HMGB1) expression on alveolar epithelial cells (A549). MethodsA549 cells were cultured and seeded at 1×10~5 cells/ml in 6-well Bioflex cell culture plates. Subsequently, the cells were exposed to cyclic mechanical stretch at 14% (group B) elongation for 4 hours using Flexercell 4000T cell stretching unit. In group C, cells were pretreated with PD98059 for 2 hours before mechanical stretch. Cells in group A without stretch were served as control. The expression of HMGB1 protein and mRNA in A549 cells were detected by immunocytochemisty staining and RT-PCR, respectively. ERK activity was measured by Western blotting method. Results Immunocytochemisty staining indicated that the expression of HMGB1 protein in A549 cells was increased obviously in group B (P<0.05) and decreased in group C (P<0.05). Polymerase chain reaction (RT-PCR) showed that the expression of HMGB1 mRNA was also significantly increased in group B (P<0.05) and decreased in group C (P<0.05). Western blotting analysis confirmed the activation of ERK in A549 cells by mechanical stretch (P<0.05). PD98059, an inhibitor of ERK, might significantly inhibit mechanical stretch induced HMGB1 protein and mRNA expression in A549 cells (P<0.05). Conclusion Mechanical stretch could regulate the expression of HMGB1 gene and protein in A549 cells through ERK signal pathway.

AIM: To screen the binding proteins to HMGB1 promoter by phage display technique. METHODS: HMGB1 promoter was incubated with phage display library. Unbound phages were eluted and phages bound to HMGB1 promoter were amplified. Twenty individual clones were randomly selected and identified by enzyme-linked immunosorbent assay (ELISA). Positive clones were characterized by DNA sequencing and the sequences were subjected for computer analysis. RESULTS: Positive phages binding to HMGB1 promoter were enriched after 4 rounds of biopanning. Twenty phage clones were selected and eleven clones of which were identified to bind specifically to HMGB1 promoter. The sequences in full length were obtained and searched for homologous sequences from GenBank. Altogether eight coding sequences were obtained, six of which were known proteins including activator protein-1(AP-1) and two of which were uncharacterized ones. CONCLUSION: Several proteins were obtained that bind specifically with HMGB1 promoter. The results will be useful for further studying the expression and regulation mechanism of HMGB1.

Objective To investigate the role of p38 MAPK pathway in the expression of high mobility group box 1 (HMGB1) in lung tissue in a rat model of ventilator-induced lung injury. Method Twenty-fonr healthy Sprague Dawley (SD) rats were randomly divided into 3 groups (n = 8 each) : group A, spontaneous breathing; group B, small tidal volume ventilation (Vt = 8 mL/kg) and group C, high tidal volume ventilation (Vt = 40 mL/kg). 1he animals in group B and C were mechanically ventilated for 4 hours and all animals were sacri-riced. The lungs were removed for: (1) lung lavage and determination of total protein contnt and WBC and neu-trophil counts in broncho-alveolar lavage fluid (BALF) ; (2) determination of W/D lung weight ratio and myelop-erexidnse (MPO) activity; (3) detennination of HMGB1 protein and mRNA expression and p38 MAPK activity in lung tissue. Differences within the groups were analyzed using One way ANOVA. Results The inflammatory re-sponse as evidenced by total protein (1.77 ± 0.68) g/L and WBC (106.55 ± 28.17) × 10~7/L in BALF, W/D lung weight ratio (7.16±1.02) and MPO activity (3.94±1.21) U/g were significantly higher in group C com-pared with group A (P <0.05); HMGB1 protein (0.64±0.17) and mRNA (1.17±0.45) expression and p38 activity (0.51±0.12) also significantly increased in group C (P <0.05). Of the above indexes, there were no statistical differences between group B and group A (P > 0.05). Conclusions High tidal volume ventilation in-daces acute lung injury, which may be related with upregulation of HMGB1 expression through p38 MAPK signal pathway.

Objective:To investigate the effect of DNA vaccine expressed Helicobacter pylori(Hp) oipA on protecting against Hp infection.Methods:The ORFs of Hp oipA had been inserted into the eukaryotic expressing vector pVAX1 and SGC-7901 cells had been transfected with recombinant plasmid pVAX1-oipA. The expression of oipA had been detected by RT-PCR and ELISA. After extracted and purified, pVAX1-oipA had been injected into BALB/c mice through muscles of right leg one time each week for three weeks(100 ?g each mouse). pVAX1 blank and normal saline had been used as the control groups. Titer of antibodies had been detected by ELISA two months after the last immunization. Based on the confirmation of immunological response in the pVAX1 groups, mice had been given orogastric challenged with live Hp Sydney strain three times(0.5?108/0.5 ml each mouse). Four weeks after challenge, mice had been sacrificed. Histological change and the colonization of Hp in the gastric mucosa had been detected by urease test, the culture of Hp, and electronic microscopy.Results:SGC-7901 cells transfected with pVAX1-oipA had expressed corresponding production at the level of transcription and translation. Immunized mice had been induced anti-oipA antibodies. After challenged with bacterium, as contrast to immunized mice groups injected with pVAX1-oipA, pVAX1 blank, normal saline, the positive rate of urease test of gastric mucosa was 0(0/10), 90%(9/10), 100%(5/5)respectively,the positive rate of cultures of Hp was 20%(2/10), 90%(9/10), 100%(5/5)respectively. Histological findings: the different degree of erosion had been observed in control group, but 80%(8/10)of gastric mucosa were normal in immunized mice.Conclusion:oipA DNA could induce effective immune response in protection against Hp infection.

To investigate the prevalence and gene types of KPC in Enterobacteriaceae strains isolated from 4 tertiary general hospitals in Hainan area ,a total 43 isolates which were resistant or intermediate to imipenem or ertapenem were collected from sterile sites between August 2012 and June 2013 from 4 tertiary general hospitals in Hainan area .Modified Hodge Tests (M HT ) were performed for KPC phenotype screening .PCR amplification and DNA sequence were performed to analyze the encoding genes of KPC .Results showed that in the 43 isolates ,21 strains were positive in M HT .PCR and DNA sequence analysis confirmed that 3 isolates produced KPC‐2 .It's suggested that there were the Enterobacteriaceae carrying KPC in Hain‐an area .The encoding genes were KPC‐2 .The KPC gene could be horizontally transmitted by plasmid among different groups of bacteria .It is important to control the transmission of these Enterobacteriaceae carrying KPC .