Animals ; Endothelium, Vascular ; injuries ; Female ; Hemodynamics ; Infusions, Intravenous ; methods ; Injections ; methods ; Male ; Rabbits ; Syringes

Objective To investigate the relationship of resistance mechanisms of a Klebsiella pneumoniae strain and a Morganella morganii strain resistance to carbapenems isolated from a single specimen. Methods Sensibility of antimicrobial agents was detected by agar dilution method. Specific PCR and DNA sequence analysis were performed to detect resistance genes. Plasmid feature was detected by plasmid conjugation and electrophoresis analysis. Genetic environment around blaKPC was analyzed with sequencing. The changes of outer membrane permeability were analyzed with electrophoresis of outer membrane proteins. Results blaKPC-2 was detected in 2 original isolates strains and their transconjugants. Carbapenem-resistance was successfully transfered by conjugation experiments. blaKPC-2 was located on dissimilar plasmids, but genetic environment around blaKPC-2 was the same sequence. The Morganella morganii isolate showed a loss of 38 ×103 OMPs and an additional 36 ×103 OMPs appearance, while the Klebsiella pneumoniae isolate showed a loss of OMPK36. Conclusion blaKPC-2 was detected in 2 isolates. This gene encoded by two plasmids with different sizes was located on the same composite transposon. The lack of outer membrane proteins could also play an important role causing isolates to exhibite resistance to carbapenems.

Objectives In order to develop a rapid microsatellite genotyping assay for inter-strain differentiation of Candida albicans isolates and understand the transmission characteristics of the infections. Methods DNA was extracted from C. albicans isolates from genitals, anal canals and oral cavities of 39 women and 27 men with genital candidiasis. The microsatellite sequences in stabel genes(CDC3, EF3 and HIS3) were amplified by a fluorescence labeled PCR. Fluorescent signals were read with an automatic se- quencer, and the data were collected with GeneScan software followed by genotyping with Genotyper soft- ware to analyze polymorphic microsatellite loci. Results Combined analysis of the 3 microsatellite markers showed 18 gene allele associations in C. albicans from genital sites of all men and women, including 10 in women, 11 in men and 3 in both. The allele associations of dominant pathogenetic strains for both sexes were 116:124, 122:131,160:200, which covered 50% of pathogenetic infection. Three common allele associations for both sexes covered 71% of all infections. Genitals and anal canals shared strains of same allele associations in 80% of women and in only 3.8% of men. The strains of same allele associations were identified in both genitals and mouth in 2.7% of women but in none of men. In their genital sites 71% of couples shared the same allele strains, of which 80% were the dominant pathogenetic strains identified in both sexes. Conclusions The improved microsatellite genotyping assay is useful for rapid differentiation, identification of infective source, and contact tracing of C. albicans infection. There are pathogenetic C. albi- cans strains with predominant allele associations in genital infections.

Objective To evaluate the screening effect of osteoporosis self-assessment tool for Asians (OSTA) in middle aged and elderly healthy men in Chengdu.Methods A total of 4042 healthy men aged 40 to 106 years received dual energy X-ray absorptiometry (DXA) assay,and OSTA index evaluation.Measurement sites included lumbar spine (L1-4),left femoral neck,trochanter,Ward's area,total hip and femoral shaft.All persons were classified into highosteoporosis-group (OSTA≤-4),mediumosteoporosis-group (-4 ＜ OSTA≤≤-1),low osteoporosis-group (OSTA＞-1),or the low risk-group (OSTA＞-1) and high risk-group (OSTA≤-1) by OSTA scores.T-scores were compared between different measurement sites detected by DXA.The sensitivity,specificity,Kappa value and the area under receiver operating characteristic (ROC) curve (AUC) of OSTA in screening osteoporosis were evaluated.Results The prevalence of osteopenia and osteoporosis in lumbar spine,proximal femur were gradually increased along with aging.The detection rate of osteoporosis in lumbar spine and proximal femur were 16.2％ and 24.0％ respectively in subjects aged over 80 years.OSTA index in low-risk,medium-risk group,high-risk group were 85.0％,11.0％,4.0％ respectively.The detection rate of osteoporosis in lumbar spine and proximal femur were 2.6％ and 1.6％ in low-risk group,10.4％ and 10.4％ in medium-risk group,and 29.3％ and 30.5％ in high-risk group,respectively.Taking OSTA ≤-1 as the cut-off value,the sensitivity and specificity of OSTA in screening osteoporosis in lumbar spine and femur by T-score＜-1 were 28.1％,28.7 ％,89.0％ and 92.4％ respectively,and by T-score≤-2.5 were 51.6％,63.2％,86.7％ and 86.8％ respectively.The consistency of diagnosis result between T-score and OSTA index according to the three versus two risk levels was 0.153 and 0.197 versus 0.195 and 0.243 Kappa value,respectively.The AUC of OSTA index for lumbar spine and femur by T-score＜-1 and T-score≤-2.5 were 0.689 and 0.823,and for different age groups and different measurement sites were 0.639 and 0.899 (all P＜0.001).Conclusions OSTA index has a certain ability in screening osteoporosis in men aged over 50 years.There are different screening results on osteoporosis among the different age groups.

To screen and identify the possible pathogen of the firstly confirmed human case of avian influenza A in Beijing, the throat swabs and tracheal aspirates of this case were collected and the H5N1 viral nucleotide was tested with real time RT-PCR. The certification of result, screening of other pathogens in respiratory tract and sub-typing of influenza viruses were made by using re-sequencing microarray. It was found that the H5N1 viral nucleic acid was positive in the tracheal aspirate of this case by means of detection with real time RT-PCR and the specific sequence of the non-structural protein (NS) gene of H5N1 virus was obtained through the detection with re-sequencing clip. Through the comparative study with the sequence in Genbank, it was proved to be the H5N1 nucleic acid of avian influenza viruses and excluded the possibility of infections with 30 subtypes of influenza viruses and 33 other respiratory tract pathogens. It is apparent that the pathogen detection with re-sequencing clip shows the high sensitivity and specificity and it plays an important role in the pathogen screening and identification for the firstly confirmed human case of avian influenza A in Beijing.

Objective To investigate cortactin expression in malignant colorectal tissues and corresponding adjacent non-cancerous colon tissues,precancerous lesions (adenomatous polyps) and the relationship between the expression of cortactin and cell division in colorectal cancer cells.Methods The expression of cortactin was detected by immunohistochemistry in colorectal cancer,colorectal adenomatous polyp (precancerous lesions) and colorectal tissues adjacent to adenocarcinomas (normal tissues).Kaplan-Meier method was employed to compare the survival between the groups.Cortactin expression and cell division were detected by Western blot and immunofluorescence in SW-620 colon cancer cells treated with cortactin siRNA.Results The positive expression rate of cortactin was significantly higher in colorectal cancer tissues than in adenomatous polyp tissues and pericarcinomatous normal tissues.Overexpression of cortactin in colorectal cancer tissues was correlated with poor differentiation (P ＜ 0.01),lymph node metastasis (P =0.006),and TNM stage (P =0.022).The 5 year survival rate of the group of negative/weak positive expression of eortactin was significantly higher than the group of strong positive expression of cortactin.CTTN gene amplification in colorectal cancer tissues was obvious.Cortactin siRNA induction caused a lower cortactin protein expression in colorectal cancer cells.Conclusions It is suggested that the excessive expression of cortactin contributes to the growth of cancer cells in colorectal cancer.

Objective To explore the effects of extract of Gingko biloba leaves (EGb) on learning, memory and hippoeampal heine oxygenase-1 (HO-1) expression in diabetic rats.Methods The behaviors of streptozotoein-induced diabetic rats were observed by Morris water maze for learning and memory after 6 months of diabetes.The HO-1 mRNA expression and protein expression in hippocampus of diabetic rats were detected by RT- PCR and immunohistochemistry respectively.Results The escape latency time in Morris water maze of diabetic rats was prolonged markedly.HO-1 mRNA and HO-1 protein expressions in hippocampus of diabetic rats were increased (1.635?0.326 vs 0.978?0.214,7.2?1.7 vs 1.9?0.5,respectively,both P＜0.01).The escape latency times in EGb (100,50 mg/kg) treated groups were shortened.HO-1 mRNA and HO-1 protein expressions in hippocampus were decreased at the same time as compared with diabetic group (100 mg/kg:0.954?0.144,2.0?0.8;50 mg/kg:0.988?0.154,2.5?0.6,all P＜0.01).Conclusion EGb can significantly inhibit HO-1 expression in hippocampus and improve learning and memory dysfunction in diabetic rats.

Objective 30 Pseudomonasaeruginosa mechanism of resistance to quinolones.Methods For the determination of ciprofloxacin MIC by agar dilution method.Used PCR on DNA gyrase and topoisomerase Ⅳ,resistance genes gyrA,gyrB, parC and parE were amplified,and BLAST,to determine whether there was resistance to bits mutation point;using pulsed-field gel electrophoresis (PFGE)of these 30 strains homology analysis.Results The 28 bacterial strains gyrA gene ampli-fied fragment of 137 points were C→T mutation causes T83I;17 strains gyrB gene amplified fragment of 351 G→C lead to G466A;parC gene amplification 21 bacteria fragment 277 point increase with C→U mutation causes S87L change two differ-ent strains parE gene locus C→U mutation A425V and A473V cause change.PFGE results:30 Pseudomonas aeruginosa could be divided into six clones,Aclone 4,B clone 7,C clone 3,D clone 14,and two other single clones.Conclusion The tar-get mutant strains closely related to the epidemic clone type,the same changes in the same pop-type strains of drug targets, and proportional to the level of ciprofloxacin MICs value,the more the number of mutated genes,MICs value higher.GyrA gene most prone to mutation,the mutation was also the first to be discovered,more than any other target of the mutation mutations on binding of drugs and targets that would be the focus of concern.

]. Conclusions The monoclonal-based ECLIA is a sensitive, specific, and rapid method and no radiocontamination. It can be used to detect hanum serum proinsulin in type 2 diabetes.

Objective To explore the changes of serum gastrin(GAS),plasma motilin(MTL) and somatostatin(SST) in critically ill children with gastrointestinal dysfunction(GID).Methods According to pediatric critical illness score,75 cases were divided into greatly critical group(score90).Fifty cases of greatly critical and critical group were divided into GID group and non-GID group.The levels of serum GAS,plasma MTL and SST were detected on an empty stomach at acute stage and convalescence stage,comparing with those of normal control group,and then,the relationship between the levels of serum GAS,plasma MTL,SST and GID,the severity of disease were analyzed.Results At acute stage,the levels of serum GAS and plasma MTL of greatly critical group and critical group were higher than those of normal control group,the levels of plasma SST of greatly critical group and critical group were lower than that of normal control group,the more severe condition of critical children,the higher level of serum GAS and plasma MTL,the lower level of plasma SST.At convalescence stage,the level of serum GAS and plasma MTL of the greatly critical group and critical group decreased and the level of plasma SST increased.The levels of serum GAS and plasma MTL of GID group were higher than those of non-GID group,but the level of plasma SST of GID group was lower than that of non-GID group.Conclusion The level of serum GAS,plasma MTL and SST can be used to assess the severity of illness and prognosis,judge the change of disease and determine the efficacy of treatment programs,and detect gastrointestinal functional lesion.