Objective To investigate the molecular epidemiology and mechanism of carbapenem resistance of Escherichia coli collected from intensive care units(ICUs)of general surgery.Methods Agardilution were carried out to confirmed the drug-susceptibility,pulsed-field gel electrophoresis(PFGE)were performed to analyze the molecular epidcmiology of carbapenem-resistance isolates.Specific PCR,DNA sequencing,conjugation experiments,plasmids extraction,plasmid transformation assays and SDS-PAGE of outer membrane proteins(OMPs)were carried to confirm genotype of carbapenemase and its transmission mechanism.Results PFGE showed the isolates belonged to 10 clonotype,and all the clinical isolates were resistant to β-lactams including imipenem and meropenem,but uncertain to aminoglycosides,specific PCR and DNA sequencing revealed that all isolates encoded carbapenem-hydrolyzing enzyme gene,KPC-2.Plasmid DNA extraction and plasmid transformation assays from some isolates comfirmed that KPC-2 encoded on a 56 kb plasmid.SDS-PAGE analysis confirmed that there are alterations in OMPs of Escherichia coli.Conclusion Escherichia coli isolates with carbapenem resistance are collected from our hospital,production of KPC-2 carbapenemase mainly contributed to reduced susceptibility of carbapenem in Escherichia coli,the alterations in OMPs may as a cofactor in high-level drug-resistance in Escherichia coli.

The aim of this paper is to apply Raman spectroscopy technique to develop rapid quantitative models for five kinds of Traditional Chinese Medicine containing CaCO3. In the experiment, Raman spectras of 67 batch of sample including Otolithum Sciaenae, Galaxeae Os, Ophicalcitum, Calcite, Stalactite and their mixture which had different content of CaCO3 were collected, and the quantitative models were established by using an improved siPLS to optimize the characteristic spectral bands and using the CaCO3 contents which were measured by EDTA titration method as references. Compared with the results by EDTA titration, the established quantitative model for CaCO, content showed a prediction result that the average relative deviation of the prediction results is 2. 71% and the average recovery rate was 100.46%, when the content is between 0.465 4-0.999 7, and when the characteristic spectral bands of 1 290-1 280, 730-714, 700-690, 660-650, 465-460, 455-445, 405-385 cm(-1) had been optimized. The result also showed that the model using Raman spectroscopy and based on an improved siPLS can get a rapid determination for contents of 5 kinds of Traditional Chinese Medicine containing CaCO3.
Calcium Carbonate ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Least-Squares Analysis ; Models, Statistical ; Plants, Medicinal ; chemistry ; Spectrum Analysis, Raman ; methods

Objective To evaluate the limit of detection of eight enzyme-linked immunosorbent assay (ELISA) according to hospital grade assessment and ISO15189:2012.Methods According to the new health industry standard WS/T 514-2017:"Establishment and verification of detection capability for clinical laboratory measurement procedures",the limit of detection (LoD) was established,in the sameset of detection system,using two reagent lot,each lot for 5 consecutive days 4 consecutive days to assess the value of the concentration of five specimens were detected repeatedly,calculated the corresponding hit rate,then transform into probability units,and the corresponding concentration value production regression model,the hit rate of 95 ％ corresponds to the probability unit 1.645 substituted into the equation,the resulting concentration value was LoD estimates.The detection limit values were tested for 3 consecutive days of detection of two LoD concentrations near the declared concentration of the sample (diluted by the standard material) was detected 4 times repeatedly to calculate the positive result was greater than or equal to the percentage of LoD statement,greater than or equal to the critical value of 87％,then verified success.Results HBsAg:0.100 IU/ml,HBsAb:9.642 mIU/ml,HBeAg:0.666 NCU/ml,HBeAb:3.700 NCU/ml,HBcAb:0.786 IU /ml,HCV:0.506 NCU/ml,TP:2.236 mIU/ml and HIV:0.135 NCU/ml.The detection limit estimates were passed.Conclusion The verification limit of the verification project in the testing method and detection system of the laboratory meet the requirements Objective.

The aim of this paper is to apply near infrared spectroscopy techniques to construct a rapid identification method for 8 kinds of mineral Chinese Medicines containing carbonates. The qualitative model using clustering analysis method in OPUS software can identify accurately 8 kinds of carbonate-containing mineral Chinese medicines. The near-infrared quantitative model was established by using partial least squares method (PLS) for 7 mineral Chinese Medicines in which main component is calcium carbonate. Compared with the results by EDTA titration, the established quantitative analysis model for calcium carbonate content showed a good prediction result that when the content is between 47.61% -99.17%, the average relative deviation of the prediction result is 0.24% and the average recovery rate was 100.3%. The results also showed that the model using near infrared spectroscopy can get not only a rapid identification of the 8 mineral Chinese medicines containing carbonates, but also an accurate and reliabe content determination of calcium carbonate for the 7 mineral Chinese medicines which contain the component.
Carbonates ; analysis ; Medicine, Chinese Traditional ; Minerals ; chemistry ; Software ; Spectrophotometry, Infrared ; methods ; Time Factors

The regulation mechanism of arecoline on rat hepatic CYP2E1 was studied in vivo. After oral administration of arecoline hydrobromide (AH; 4, 20 and 100 mg x kg(-1) x d(-1)) to rats for one week, the hepatic CYP2E1 mRNA level remained unchanged, but the hepatic CYP2E1 protein content was dose-dependently increased. Additionally, although the hepatic CYP2E1 activity was induced by AH treatment, the induction was attenuated with the increase in dosage. The results indicate that the effect of arecoline on rat hepaticdoes not involve transcriptional activation of the gene, but largely involves the stabilization of CYP2E1 protein against degradation or increased efficiency of CYP2E1 mRNA translation, and additionally involve the post- ranslational modification of CYP2E1 protein. Furthermore, the CYP2E1 response is fairly equal among the different species, the induction of rat hepatic CYP2E1 by arecoline suggests that there is a risk of metabolic interaction among the substrate drugs of CYP2E1 in betel-quid use human.
Animals ; Arecoline ; pharmacology ; Cytochrome P-450 CYP2E1 ; metabolism ; Cytochrome P-450 CYP2E1 Inducers ; pharmacology ; Humans ; Liver ; drug effects ; metabolism ; RNA, Messenger ; Rats

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.

Background Sirius system,a new Scheimpflug camera combined with Placido topography,improved the capability of imaging the anterior eye segment significantly.However,the study of assessing the repeatability and agreement between Sirius and Pentacam is still lack up to now.Objective This study was to evaluate the repeatability and agreement of the anterior ocular segment measuring parameters by Sirius and Pentacam in myopia received laser in situ keratomileusis (LASIK).Methods Thirty-five myopic eyes of 35 patients received LASIK were included in School of Optometry and Ophthalmology Eye Hospital from 2010 May through 2010 July.Corneal power flat keratometry (Kf),step keratometry (Ks),mean keratometry (Km),thinnest corneal thickness(TCT),the location of TCT,anterior chamber depth (ACD) and anterior chamber volume (ACV) were measured by Sirius and Pentacam in all the eyes,respectively.The repeatability of the measuring results were evaluated using intraclass correlation coefficients (ICC) and Cronbach's coefficient alpha (CoA),and the agreement of measuring parameters between Sirius and Pentacam was analyzed using Bland-Altman plot.Results Both Sirius and Pentacam demonstrated high intraobserver repeatability,with all ICC and CoA more than 0.90.No significant differences were found in Kf values and Ks values between the two methods (t =-1.533,-1.750,P＞0.05).Km value was (39.14 ± 1.95) D by Sirius measurement,which was sígnificantly higher than (39.05 ± 1.91) D by Pentacam measurement (t =3.572,P =0.001).The TCT was (457.6 ± 40.9) μm by Sirius method,showing a significant reduce in comparison with (465.4±37.5) μm of Pentacam method (t =-6.689,P＜0.001).A positive correlation was seen in the TCT between the two methods (r=0.988,P＜0.001).The Bland-Alrman plots showed the 95％ CI-21 μm to 6 pm in the TCT value between the two devices.Pairwise comparison of the location of TCT measurements showed significant differences between the two devices (t =-4.132,-5.696,P＜0.001),with a good correlation (r=0.751,0.775) and the 95％ CI (-0.36-0.17 mm,-0.35-0.12 mm).A very good agreement was seen in ACD between the two devices (-0.02-0.12 mm),but the agreement result was not very well in the ACV between the two devices with the 95％ CI (-27.70-6.20 mm3).Conclusions Sirius and Pentacam measurements for anterior ocular segment parameters have a very good repeatability in post-LASIK eyes.In addition,good agreement results are exhibited in corneal power,TCT and ACD between Sirius and Pentacam with an acceptable maximal different value between them.Sirius and Pentacam can be used interchangeably in clinical examination.However,the two devices can not interchangeably for ACV measurement and TCT location.

Objective To explore the accuracy of Vitek 2 compact advanced expert system (AES) in indicating and analyze the carbapenemases-resisting Enterobacteriaceae phenotypes,and further investigate the methods to make up the AES.Methods 28 Enterobacteriaceae strains with Imipenem-Nonsusceptible by Vitek 2 compact,but AES suggested all production of carbapenemases were isolated.And imipenem susceptibility was determined by the disk diffusion method.Modified Hodge test (MHT) and the metallo-β-1actamase was detected by the double disk synergy method.Resistance genes were detected by the PCR amplification.Results ESBLs gene was amplified from all 28 selected strains,16 of which was detected KPC gene,and no strain of metallo-β-1actamases-producing bacteria.With carbapenemase gene detection as the gold standard,the accuracy of AES was 57.1％.Disc diffusion method detection accuracy rate of imipenem was 100％,and for 100％ of MHT accuracy.PCR amplification,MHT and the disk diffusion displayed the same result in detecting carbapenemases,but different with AES (x2 =10.08,P＜0.05).Conclusion The indications of the presence of carbapenemases using AES was not completely correct with a certain false-positive,and it is necessary to take other methods,such as disk diffusion or MHT methods,and improve the reliability of medicine-sensitivity tests.