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.

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

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.

]. 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.

Background Biometry of the anterior ocular segment parameter is very important for the diagnosis and treatment of glaucoma and ocular injury as well as measurement of intraocular lens(IOL).Objective This study was to compare the differences in the anterior chamber depth(ACD) and the central corneal thickness (CCT) between Sirius and Pentacam and evaluate the agreement of these two measurement methods.Methods The ACD and the CCT of 38 right eyes from 38 health volunteers aged 23- 32 years were measured with both Pentacam and Sirius.Three times of measurement were pedormed on each eye for each method to obtain the average values.The repeatability and agreement from each method were assessed as intraclass correlation coefficient( ICC ) and coefficient of variation(CV) and the agreement between these two methods were evaluated using Bland-Altman mode.ResultsThe mean ACD value was( 3.18±0.21 ) mm from Pentacam with the ICC 0.995 and CV 0.066.The mean ACD value from Sirius was (3.22 ±0.21 )mm with the ICC 0.996 and CV 0.065.The difference value in ACD between two methods was 0.04 mm,showing a significant difference( t =-6.225,P＜0.05 ) and a positive correlation (r=0.977) between two methods.The 95％ limit of agreement was( -0.04-0.13)mm within 1 standard difference (SD) of the mean value( ±0.21mm),which was acceptable for clinical measurement.The CCT was( 535±33 )μm from Pentacam with the ICC 0.994 and CV 0.062.The CCT was(537±36)pm from Sirius with the ICC 0.999 and CV 0.067.The difference value in the CCT between two methods was about 2 μm,presenting a in significant difference ( t =1.771,P＞0.05 ) and positive correlation ( r =0.985 ).The 95 ％ limit of agreement was ( - 11.64-15.65 ) μm within 1 SD of the mean value( ±34.27 pm),which was acceptable for clinical measurement.ConclusionsSirius and Pentacam show good agreement in the measurement of ACD and CCT.The two methods offer an alternative choice for the biological measurement of the anterior ocular segment.

Background Axial length and anterior chamber depth are important parameters for the calculation of diopter of intraocular lens ( IOL ). Objective This study was to investigate and compare the measuring outcomes of axial length and anterior chamber depth with IOLMaster,Axis- Ⅱ A-scan and ODM 1000A sonograph.Methods This a observational study.Axial length and anterior chamber depth were measured in 83 eyes of 48 patients with IOLMaster,Axis-Ⅱ A-scan and ODM 1000A sonograph by the same operator.The measuring results were compared among the three methods.Results The axial length were(25.79±0.85) mm,(25.72± 0.82 )mm and ( 26.00 ±0.83 )mm respectively with Axis- Ⅱ,ODM 1000A sonograph and IOLMaster.The difference between Axis-Ⅱ and DM 1000A sonograph was (0.07 ± 0.35 )mm without statistical difference between them (t=1.711,P =0.091 ).The difference of axial length between IOLMaster and DM 1000A sonograph was ( 0.27 ±0.29) mm with a statistical difference between them ( t =-8.570,P =0.000 ).The difference between IOLMaster and Axis- Ⅱ was (0.21 ±0.32 ) mm and showed a statistical difference ( t =- 5.931,P ＜ 0.01 ).The positive correlations were found in the axial length values by the each other comparison among the three instruments( r=0.916,0.938,0.928,P＜0.01 ).The anterior chamber depth values were ( 3.81 ±0.21 ) mm,( 3.84 ±0.25 ) mm and ( 3.83 ±0.18 )mm respectively with Axis-Ⅱ,0DM 1000A sonograph and IOLMaster.The difference of anterior chamber depth between Axis- Ⅱ and DM 1000A was (0.03 ±0.17 ) mm without statistical difference between them ( t =- 1.324,P =0.189 ).The difference in the anterior chamber depth between IOLMaster and DM 1000A was (0.01 ±0.15 ) mm and that between IOLMaster and Axis-Ⅱ was( 0.01 ±0.12)mm without any statistical differences among them (t =0.815,P=0.417 ;t=-0.900,P=0.371 ).The high correlation between anterior chamber depth measurements were found by the each other comparison in the three instruments ( r =0.735,0.813,0.823,P ＜ 0.01 ).Conclusions ODM 1000A sonograph can provide precise axial length and anterior chamber depth values.However,ODM 1000Asonograph can not substitute for IOLMaster in the measurement of the anterior chamber depth and axial length.

Background Clinical research showed that the corneal endothelial cell density value from different corneal specula microscopies exist diversity.The relevant literature of SP02,Tomey EM-3000 and SP3000P is still seldom up to now. Objective This research was to assess the repeatability of endothelial cell density measurements by SP02,Tomey EM-3000 and SP3000P respectively and the agreement among 3 kinds of endothelial microscopes.MethodsFifty-four healthy volunteers with the age 17-38 years old were included this research.The written informed consent was obtained from each subject before examination.The corneal endothelial cell densities in the right eyes were analyzed with SP02,Tomey EM-3000 and SP3000P respectively for 3 times under the automatic mode,and the analytical procedure of SP3000P measurement were divided into automatic mode SP3000P (A) and manual correction modes SP3000P( M).The repeatability of each specula microscopy was analyzed by calculating the intraclass correlation coefficients (ICC) and coefficient of variation ( CV ),and the 95％ confidence intervals and plotting Bland-Altman graphs were used to analyze the agreement among these methods.ResultsThe mean corneal endothelial cell densities in the population ＜24 years were significantly higher than the ones ≥ 24 years (t =3.692,P＜0.05 ),but no statistical difference was found between different gender ( t =0.335,P =0.739 ).The mean corneal endothelial cell densities were ( 3058 ± 260 ),( 2954 ± 229 ),( 2668 ± 258 ),( 2734 ± 268 ) cell/mm2 ; the ICCs were 0.957,0.940,0.972 and 0.972 and the CV were 0.063,0.061,0.056,0.058 for SP02,Tomey EM-3000,SP3000P (A) and SP3000P ( M ) respectively.The 95％ confidence intervals were ( - 100.8 - 306.8 ),( 162.6 - 617.4 ),( 109.9-494.1 ) and ( -0.6 - 132.6 ) cell/mm2 for between SP02 and Tomey EM-3000,SP3000P ( A ) and SP02,SP3000P(A) and Tomey EM-3000,SP3000P(A) and SP 3000P(M) respectively.ConclusionsSP02,Tomey EM-3000 and SP3000P(A) have good repeatability in the measurement of corneal endothelial cell density,however the outcome is different.Therefore,it is not interchangeable for the detection of corneal endothelial cell density.The differences of corneal endothelial cell density obtained from these instruments shall be paid high attention for their differences.SP3000P(A) and SP3000P(M) can be used interehangeably and SP3000P(A) is a preferable choice due to its convenience and quickness.

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 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.