Objective:To explore the effect and mechanism of AMPK on apoptosis of alveolar epithelial cells induced by endoplasmic reticulum stress in COPD rats.Methods:the rats were divided into three groups:control group,model group,AICAR intervention group,establishment of rat model of chronic obstructive pulmonary disease by smoking smoke inhalation and intratracheal instillation of lipopolysaccharide.The HE staining of rat lung tissue pathological observation,immunohistochemical detection of p-AMPK /AMPK,western blot the expression of Caspase-3,ORP 150,and CHOP.Apoptosis were detected by TUNEL method.Results:the HE staining showed that the model group of pulmonary bullae formation,inflammatory cell infiltration,inflammatory ceils in AICAR group was lower than that of model group.Compared with the normal control group,immunohistochemistry and Western blot showed that p-AMPK/AMPK and ORP150 protein expression decreased in the model group,the difference was statistically significant (P＜0.05),and AICAR in the intervention group p-AMPK/AMPK and ORP150 protein expression were significantly increased compared with the model group,the difference was statistically significant (P＜0.05).Endoplasmic reticulum stress related apoptosis The expression of CHOP and caspase-3 apoptosis index increased significantly in the model group,there was significant difference compared with normal group (P＜0.05),while in group AICAR,apoptosis index down significantly compared with the model group.Conclusion:AMPK can protect alveolar epithelial cells from cigarette smoke induced endoplasmic reticulum stress and apoptosis,it was possible to achieve its protective effect the increase of ORP150.

Objective To establish a method for detecting Aspergillus spp.by Loop-mediated isothermal amplification.Methods Aspergillus spp.specific primers were designed from the relative conservation region of the published sequence of 28S rRNA genes of A.fumigatus (GenBank accession number AY660917),A.terreus (GenBank accession number AF454183),A.flavus (GenBank accession number AF454158),and A.niger (GenBank accession number AF454169).Genomic DNA were extracted from Aspergillus standard strains,clinical control strains and clinical samples,and amplified by LAMP.The amplified results could be read with the naked eye by the coloring effect of fluorescent nucleotide dye without the DNA electrophoresis.Approximately 103 each Aspergillus conidia suspension was added to the serum from healthy volunteers,and detected these simulative clinical samples bv LAMP assay.Results The LAMP and PCR assays obtained positive results for all four Aspergillus species and 8 simulative clinical samples (double samples for each Aspergillus species) including 103 conidia,but negative in the remaining 15 non-Aspergillus species,human total blood genomic DNA,30 clinical serum samples infected with non-Aspergillus and 10 healthy volunteers.The LAMP assay had a minimum detection of 0.05-0.5pg,by means of detect different levels of 500,50,5,0.5,0.05 pg template in each reaction tube.Conclusion The results confirm that LAMP is a simple,rapid,sensitive and specific method,and can be used for detection of Aspergillus strains in clinical and environmental specimens.

Objective To explore the application and analysis of E-PASS scoring system for orthopaedic senile hip surgery risk assessment. Methods Retrospectively analyzed the data of 188 cases of elderly hip fracture patients with anesthesia and surgery, postoperative complications. Useing E-PASS scoring system to evaluate the mortality of patients with surgical risk. Results Postoperative complications mainly was respiratory system diseases, the rate of complication was 25.88%(22/85), accounting for total cases rate was 11.70% (22/188). Secondly for cardiovascular diseased and urinary system diseases. E-PASS scoring system showed that the scores of surgery risk assessment in patients with complication was significantly higher than that without complication:(0.27±0.23) scores vs. (0.17±0.16) scores, t=3.728, P<0.05. Ninety-one patients were evaluated to occur complications by E-PASS grading system, 85 patients occurred practically,the rate of actual value/forecast was 0.93.Six patients were evaluated to die, 2 patients occurred practically, the rate of actual value/forecast was 0.33, there were no significant differences ( P=0.056,0.124). Conclusion E-PASS scoring system for orthopaedic senile hip surgery risk assessment can predict the risk of complication and death effectively,it is value of interventing the preoperative risk factors and has great clinical application value.

Objective To investigate the distribution of virulence-related genes in multidrug resistant Escherichia coli.Methods Seven virulence genes papA,cnf1,cnf2,cfaB,ipaB,hofQ and ompT were detected by PCR in 20 strains of multidrug resistant Escherichia coli clinically isolated,and the positive genes were further searched in 31 strains of Escherichia coli in BioCyc database whose genomies had been fully sequenced.Results Virulence genes hofQ and ompT were detected in 20 strains of Escherichia coli with a positive rate of 95.0％ (19/20) and 55.0％ ( 11/20),respectively.Among 31 strains of Escherichia coli in BioCyc,21 (67.7％) were positive for hofQ gene and 15 (48.4％) were positive for ompT gene.Conclusion hofQ and ompT genes are prevalent in multidrug resistant Escherichia coli.

OBJECTIVE To investigate the antibiotic resistance of multi-resistant Acinetobacter baumannii(ABA) and distribution of aminoglycoside-modifying enzymes and 16S rRNA methylase genes in ICU in Yinzhou People′s Hospital in Ningbo. METHODS The samples of 20 ABA isolates were collected from Oct 2007 to Jul 2008 in ICU.K-B method was used to determine the sensitivity to 32 antibacterials and the aminoglycoside-modifying enzymes and 16S rRNA methylase genes were analyzed by polymerase chain reaction(PCR). RESULTS From 20 ABA isolates,8 strains carried aminoglycoside-modifying enzymes genes aac(3)-Ⅰ,aac(3)-Ⅱ,aac(6′)-Ⅰb,and ant(3″)-Ⅰ,their positive rate was 10%,15%,30% and 25%,respectively;no strain carried 16S rRNA methylase genes. CONCLUSIONS The antibiotics resistance of A.baumannii is very serious in Yinzhou People′s Hospital in Ningbo.Aminoglycoside-modifying enzymes genes aac(3)-Ⅰ,aac(3)-Ⅱ,aac(6′)-Ⅰb and ant(3″)-Ⅰ exist in multi-resistant A.baumannii widely.They would be the main causes of high drug-resistantce to aminoglycosides.

OBJECTIVE To investigate the antibiotics resistance of multi-resistant Acinetobactor baumannii(ABA) and genotypes of beta-lactamases in ICU.METHODS The samples of 20 A.baumannii isolates were collected from Oct 2007 to Jul 2008 from patients in ICU.To determine the sensitivity to the 32 kinds of antibacterials,K-B method was used and the detection of ESBLs and AmpC beta-lactamases was performed by three dimensional test and 21 types of beta-lactamases genes were analyzed by polymerase chain reaction(PCR).RESULTS Among the 20 ABA isolates,all carried TEM beta-lactamases gens(100%),10 carried OXA-23 beta-lactamases gens(50%) and 15 strains carried ADC beta-lactamases gene(75%),50% strains produced TEM,OXA-23 and ADC beta-lactamases simultaneously.Through determining sequence of one PCR product from TEM,OX23 and ADC respectively,we found TEM-116,OXA-73 and ADC-25 type beta-lactamases genes.CONCLUSIONS The antibiotics resistance of ABA is very serious.TEM,OXA-23 and ADC exist in multi-resistant A.baumannii widely.It should be the main causes as high rate of drug-resistantce to beta-lactamantibiotics.

OBJECTIVE To investigate the antibiotics resistance of Escherichia coli(ECO) and distrubtion of aminoglycoside-modifying enzymes and 16S RNA methylase genes in ICU.METHODS The samples of 20 ECO isolates were collected from Dec 2007 to Jun 2008 of patients in ICU.To determine the sensitivity to the 30 antibacterials K-B method was used and aminoglycoside-modifying enzymes and 16S RNA methylase genes were analyzed by polymerase chain reaction(PCR).RESULTS In among the 20 ECO isolates,8 strains carried aac(3)-Ⅱ(40%),3 carried aac(6′)-Ⅰb(15%) and ant(3″)-Ⅰ(15%),1 be found aph(3′)-Ⅰ(10%)and 13 be found aadA4/5 aminoglycoside-modifying enzymes genes,no strain carried 16S RNA methylase genes.CONCLUSIONS aac(3)-Ⅱ、aac(6′)-Ⅰb,ant(3″)-Ⅰ,aph(3′)-Ⅰ and aadA4/5 aminoglycoside-modifying enzymes genes exist in ECO widely,they should be the main cause inducing the high rate of drug-resistance to aminoglycosides.

OBJECTIVE To study the mechanisms of chromosome and plasmid-mediated quinolones resistance in Escherichia coli(ECO).METHODS Clinical isolates of ECO were collected from clinical specimens at ICU of Yinzhou People′s Hospital from Dec 2007 to Jun 2008.To detect the susceptibility to 30 types of antibiotics K-B disk diffusion method was used.The susceptibicity to ciprofloxacin and levofloxacin were detected by agar dilution testing.Then six type genes gyrA,qnrA,qnrB,qnrS,aac(6′)-Ⅰb-Cr,and qepA were investigated by PCR.In the meantime,the PCR products were sequenced.RESULTS The alterations in gyrA were found in all 20 tested strains.Two new subtypes were found in ECO 13 and ECO 15.Three strains were found acc(6′)-Ⅰb-Cr and ECO 2 was detected qurA gene.CONCLUSIONS The mutations in gyrA play a dominant role in the resistance to quinolones in ECO.Both aac(6′)-Ⅰb-Cr and qepA may responsible for quinolones resistance in ECO too.

OBJECTIVE To investigate mycoplasma infection in ICU patients.METHODS Sixty-five samples from blood,respiratory tract and genitourinary tract of patients were collected respectively from Oct 2007 to July 2008 in ICU.Mycoplasma pneumoniae(Mp),Urealasma urealytium(Uu) M.fermentans(Mf) and M.penetrans(Mpe) were cultivated by modified mycoplasma fluid and solid medium.Mf and Mpe positive isolates were verified by nested polymerase chain reaction(rPCR),Mp and Uu were confirmed by fluorescent quantitative PCR.RESULTS It was found that the positive detection rate for Mp was 12.3%(8/65)in blood and 35.4%(23/65) in respiratory tract excreta and for Mu 1.5%(1/65) and 26.2%(17/65) in blood or Genitourinary tract,respectively.Mpe and Mf did not detected.CONCLUSIONS The state of mycoplasma infection is very severe,and often accompanies bacterial infection.It is necessary to consider mycoplasma when chose antibiotics.

Objective To characterize the changes in the dimensions during systolic and diastolic periods in the aorta with ECG-gated multi-detector CTA scans.Methods The CT angiograms of 115 patients (78 males,mean age 55.2 ± 9.4 years;37 females,mean age 60.1 ± 8.5 years) both in systolic and diastolic periods were obtained on a 64-slice ECG-gated multi-detector CT scanner.The diameters were measured at four anatomic levels of the aorta.(Level A:1 mm proximal to the innominate artery;Level B:1 mm distal to the left common carotid artery;Level C:1 mm distal to the left subclavian artery;Level D:10cm distal to the left subclavian artery).On each level,the maximal and the minimal diameters were measured both in systolic and diastolic periods.Results The paired sample t test results showed a significant difference between the systolic and diastolic diameters in all individual subjects on every level (P ＜0.001).The mean maximum diameter changes were 1.95％ (range-2.0％ to 7.0％),2.12％ (range-3.0％ to 6.0％),1.88％(range-1.0％ to 8.0％)and2.47％(range-3.0％ to 10.0％)at level A,B,C and D,respectively.The mean minimum diameter changes were 1.43％ (range-3.0％ to 5.0％),2.67％ (range-2.0％ to 11.0％),1.75％ (range-14.0％ to 9.0％)and 2.99％ (range -2.0％ to 11.0％) at level A,B,C and D,respectively.Conclusions The differences of the aortic diameters between systolic and diastolic periods are significant.The pulsatility of aorta in Chinese population may be different from published Western literature.