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.Pharmacological Study on Periplaneta Americana against Alcohol-induced Liver Injury
Chenchen LI ; Yong'ai XIONG ; Chengjia TAN ; Jing HUANG ; Xiaodong JIANG ; Ming MEI ; Li HAN
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(4):758-761
This study was aimed to explore the effect of Periplaneta Americana against alcohol-induced liver injury in mice, in order to provide a theoretical basis for the industrial production of P. Americana. Kunming mice were randomly divided into six groups, which were the normal group, model group, positive group, high-, middle-, low-dose of P. Americana groups. Intragastric administration of Tiopronin Enteric-coated Tablets 100 mg·kg-1 was given to the positive group. Intragastric administrations of whole powder of medicine were given to the high-, middle-, low-dose groups with the dosage of 6.667, 3.333, 1.667 g·kg-1, respectively. The drugs were given daily for 10 con-secutive days. After 3h of the 10th day drug administration, intragastric administration of distilled water was given to the normal group, while 14mL·kg-1 of 56℃ Red Star Liquor was given to other groups. No food was given but water for 12h. Blood was collected from the orbit. The ALT, AST and GGT in blood serum of mice were measured. The liver was dissected and liver coefficient was calculated. Histopathological examination was given on liver tissues. The results showed that compared with the normal group, the level of ALT, AST in blood serum of the model group had obvious enhanced (P< 0.01), the level of GGT had obvious enhanced (P< 0.05). Compared with the model group, the level of GGT, AST of the high-, middle-dose group had obvious enhanced (P< 0.05), the level of ALT activity had obvious enhanced (P< 0.01). There were severe liver histopathological damages in mice of the high-, middle-, low-dose group. It was concluded that P. Americana had some side effects in the treatment of alcohol-induced liver injury.
5.The influence of body temperature on the recovery from vecuronium-induced neuromus cular block
Hai-Ming WEN ; Wen-Qi HUANG ; Dong-Mei DAI ; Al ET
Chinese Journal of Anesthesiology 1994;0(03):-
Objective To investigate the influence of body temperature on the recovery from vecuronium-induced neuromuscular block.Methods Sixty-eight ASA I - II patients (39 male, 29 female) aged 19-69 yr undergoing elective surgery under general anesthesia were randomly divided into 2 groups: group I in which patients' body temperature was maintained at 37 ℃ using warming blanket; group II in which no measures were taken to maintain the patients' body temperature. The patients were premedicated with phenobarbital 2 mg?kg-1 and atropine 0.01 mg? kg-1 intramuscularly. Anesthesia was induced with fentanyl 5 ?g? kg -1, propofol 2 mg? kg-1 and vecuronium 0. 1 mg?kg-1 . After tracheal intubation anesthesia was maintained with inhalation of 0.8%-2.5% isoflurane and propofol infusion at a rate of 2-4 mg ? kg-1? h-1 .Neuromuscular block was monitored using accelograph (Biometer, Denmark) .The changes in TOF and T1 were monitored. T1was maintained at 10% by vecuronium infusion during operation. At the end of operation a bolus of vecuronium 80?g ? kg-1 was given intravenously and T1 was completely depressed. The time for T1 to returned to 5% ,25% and 90% and the time required for T1 to return from 25 % to 75 % were recorded. The total amount of vecuronium given was recorded. Temperature probe was placed in the esophagus ( core temperature) . The room temperature was also recorded. Results The body temperature was lower, the total dose of vecuronium was smaller and the vecuronium-induced neuromuscular block lasted longer in group II as compared with group I . There was close correlation between body temperature and vecuronium-induced neuromuscular block. Conclusions Lower core body temperature could prolong the vecuronium-induced neuromuscular block.
6.ISOLATION OF ACIDOPHILIC AND ACIDODURIC STREPTOMYCETES USING DISPERSION AND DIFFERENTIAL CENTRIFUGATION APPROACH
Li-Ming WANG ; Ying HUANG ; Qing-Feng CUI ; Qiong XIE ; Ya-Mei ZHANG ; Zhi-Heng LIU ;
Microbiology 1992;0(06):-
Technological improvement for microorgnism isolation is important since isolation provides substantial materials for the exploitation of new microbial resources. In this study, a new approach, dispersion and differential cetrifugation (DDC), was applied in the isolation of acidophilic and acidoduric streptomycetes from 12 acid soil samples. Contrast with traditional method, the new approach yielded satisfying results with 2 - 20 times isolation efficiency and good selectivity. 45 representatives out of 249 streptomycetes isolates, which belonged to 12 color groups, showed morphology and cell wall type consistent with streptomycetes. The optimum pH range for their growth were between pH 4.5 - 5.5. It is proved that we succeeded in the rare-streptomycetes isolation using DDC approach.
7.Development and Application of X-Cluster: a New Software for Numerical Classification
Ji-Xiang HUANG ; Ming HUI ; Dong-Mei QI ; Tian-Gui NIU ;
Microbiology 1992;0(01):-
To remedy the limitations of traditional numerical classification softwares,a new application,X-Cluster,was developed by using various design patterns.X-Cluster had powerful functions to support the researching of numerical classification,and testified by some classify studying about Bacillus spp..
8.The Techniques of Isolation and Determination of Ergosterol as the Indicator of Fungal Biomass
Xing-Mei XI ; Guang-Ming ZENG ; Hong-Yan YU ; Jian-Bing LI ; Guo-He HUANG ;
Microbiology 1992;0(03):-
Ergosterol is the important component of the fungal membrane, and having stable structure. This makes it a suitable indicator for growth of fungi. In the paper, isolation and determination techniques of ergosterol as the indicator of the fungal biomass were reviewed. The methods of extracting ergosterol include traditional saponification and refluxing, rapid physical disruption, rapid ultrasonication, supercritical fluid extraction and so on. The ergosterol determination methods are high performance liquid chromatography, gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, and thin-layer chromatography, et al. The application of these techniques was also introduced. Finally, the paper prospected the feasibility of applying the ergosterol as the indicator of fungal biomass in composting.
9.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.
10.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.