1.Study on the molecular mechanism of carbapenem-resistant Escherichia coli
Shougang KUAI ; Haifeng SHAO ; Weiping WANG ; Ming FAN ; Mei HUANG
Chinese Journal of Microbiology and Immunology 2010;30(9):829-833
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.
2.Quantitative models of Raman spectroscopy for five kinds of traditional Chinese medicine containing CaCO3 based on an improved siPLS.
Long CHEN ; Ming-yang YUAN ; Jing MING ; Yi-mei LIU ; Bi-sheng HUANG ; Ke-li CHEN
China Journal of Chinese Materia Medica 2015;40(18):3608-3615
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
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chemistry
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Drugs, Chinese Herbal
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chemistry
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Least-Squares Analysis
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Models, Statistical
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Plants, Medicinal
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chemistry
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Spectrum Analysis, Raman
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methods
3.Establishing and Verifing Detection Limit for the Eight Items before Blood Transfusion with Enzyme-Linked Immunosorbent Assay by the WS/T 514-2017 Industry Standard Documents
Wei-Ming CEN ; Bao-Huan LIANG ; Ming-Lan HUANG ; Yuan-Ping HUANG ; Yue-Mei LIANG ; Feng-Mei FANG ; Hong SUI
Journal of Modern Laboratory Medicine 2018;33(1):141-144
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.
4.A NIR qualitative and quantitative model of 8 kinds of carbonate-containing mineral Chinese medicines.
Ming-Yang YUAN ; Bi-Sheng HUANG ; Chi YU ; Yi-Mei LIU ; Ke-Li CHEN
China Journal of Chinese Materia Medica 2014;39(2):267-272
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
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analysis
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Medicine, Chinese Traditional
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Minerals
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chemistry
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Software
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Spectrophotometry, Infrared
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methods
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Time Factors
5.Discussion on the Problems of Vitek 2 Compact Advanced Expert System to Identify Carbapenemase Phenotypes in Isolates of Enterobacteriaceae
Ying WANG ; Fangfang CHEN ; Mei HUANG ; Haiyan XI ; Ming FAN ; Haifeng SHAO ; Weiping WANG
Journal of Modern Laboratory Medicine 2017;32(4):101-103,106
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.
6.Analysis of effect of universal salt iodization on iodine nutrition among children aged 8~10 years in iodine deficiency area in Shandong Province
Ju-mei, HUANG ; Jin-biao, WANG ; Xiao-ming, WANG ; Wen, JIANG ; Yuan, LIU
Chinese Journal of Endemiology 2008;27(5):543-544
Objective To explore the status of iodine nutrition in 8~10 years children after universal salt iodization(USI)in the iodine deficiency area.Methods Probability proportion to size method(PPS)or simple random sampling methods were used to sample children aged 8~10 years in iodine deficiency area in the vear 1995,1997,1999,2002 and 2005, respectively.Goiter were detected by palpation and B-ultrasound, iodine concentration in salt was detected by direct titration method and that in urine by the method of As3+-Ce4+catalytic spectrophotometry.Results After USI has been implemented,the median of salt and urinary iodine tended to mcreaseand the goiter rate tended to decrease year by year.In 2005,the coverage rate of iodinated salt was elevated to 97.2%,qualified iodize salt rate was 97.1%and edible qualified iodinated salt rate was 94.3%in the whole iodine deficiency areas.The median of urinary iodine Was 227.7 μg/L 89.7%(323/360)of the population had a level of the urinary iodine over 100μg/L Goiter rate of 8~10 years children Wag decreased from 22.3%(282/1267)to 4.4%(53/1200) from 1995 to 2005.Conclusion After 10-year USI,the status of iodine nutrition in ShaJldong Province has been promoted obviously and it is in a suitable iodine nutritional status.
7.Predictability of multi-slice CT perfusion in the restorability of renal function of hydronephrotic kidneys
Hui YE ; Dao-Yu HU ; Qia-Xia WANG ; Ming XIAO ; Wen-Hua HUANG ; Jin-Mei SONG ;
Chinese Journal of Radiology 2001;0(04):-
Objective To evaluate the predictability of MSCT perfusion in the restorability of renal function of hydronephrotie kidneys with unilateral partial ureteric obstructed rabbit model as to explore a method to predict the restorability of renal function of hydronephrotic kidneys and to investigate the changes of MSCT perfusion parameters during the course of the restore of renal function.Methods Establish a unilateral partial ureteric obstructed rabbits hydronephrotie model.Hydronephrotie rabbits were grouped as control,2,4 and 8 week(G_2w,G_4w and G_8w)after obstruction and the later 3 groups of rabbits were reared for further 4 weeks after the obstruction was released.MSCT perfusion scanning was performed and the specimen was made into histological slices with HE staining.Results BF and BV value of renal cortex and medulla of G_2w after obstruction [(864?32)ml?100 g~(-1)?min~(-1),(19.5?0.9)ml/100 g (cortex ); (182.1?7.5)ml?100g~(-1)?min~(-1),(8.37?0.51)ml/100g(medulla)]was released restored in substance and approached that of control[(899?63)ml?100g~(-1)?min~(-1),(21.6 + 1.4)ml/100 g (cortex);(193.5?16.5 )ml?100g~(-1)?min~(-1),(8.50?0.54 )ml/100 g (medulla)]while there was no significant restore in that of G_4w and G_8w after obstruction[(525?15)ml?100g~(-1)?min~(-1),(12.8? 0.6)ml/100g (G_4 w);(512?10)ml?100g~(-1)?min~(-1),(9.4?1.0)ml/100 g (G_8w)] was released. Histologically,there was a positive correlation between the duration of obstruction and the seriousness of pathologic changes.Conclusion MSCT perfusion can provide information not only morphologically but also about renal perfusion of hydronephrotic kidneys.
8.Induction of rat hepatic CYP2E1 expression by arecoline in vivo.
Xiang-tao HUANG ; Run-mei XIAO ; Ming-feng WANG ; Jun-jun WANG ; Yong CHEN
Acta Pharmaceutica Sinica 2016;51(1):153-156
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
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Arecoline
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pharmacology
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Cytochrome P-450 CYP2E1
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metabolism
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Cytochrome P-450 CYP2E1 Inducers
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pharmacology
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Humans
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Liver
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drug effects
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metabolism
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RNA, Messenger
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Rats
9.Comparison of anterior chamber depth and central corneal thickness measuring values between Sirius and Pentacam
Yan, LI ; Shi-ming, CHENG ; Xin, YANG ; Jin-hai, HUANG ; Qin-mei, WANG
Chinese Journal of Experimental Ophthalmology 2012;30(3):262-265
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.
10.Comparison of axial length and anterior chamber depth measurements by IOLMaster, Axis- Ⅱ A-scan and ODM 1000A sonograph
Shi-ming, CHENG ; Jin-hai, HUANG ; Yan, LI ; Xin, YANG ; Qin-mei, WANG
Chinese Journal of Experimental Ophthalmology 2011;29(11):1023-1026
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.