2.Investigation of ?-lactamase genotypes of Chryseobacterium/Flavobacterium spp.
Rong ZHANG ; Hong-Wei ZHOU ; Jie DONG ; Gong-Xiang CHEN ;
Chinese Journal of Laboratory Medicine 2001;0(03):-
Objective To understand the distribution of the genotypes of ?-lactamases in Chryseobacterium/Flavobacterium spp.Methods Minimum inhibitory concentrations (MICs) of 43 Chryseobacterium meningosepticum strains,22 Chryseobacterium indologenes strains and 10 Chryseobacterium gleum strains against 15 antibiotics were determined by agar dilution method.3-D test and modified 3-D test were used to identify carbapenamase.2-mercaptopropionic acid inhibitory test was used to confirm metallo-?-lactamases (MBL).Genes of ?-lactamases were amplified with 6 pairs of primers special for Chryseobacterium/Flavobacterium spp.and the amplified genes were sequenced.Results MIC_(50) and MIC_(90) of quinolones were lower comparing to other antibiotics.MICs of C.gleum against 15 antibiotics were lower than other Chryseobacterium/Flavobacterium spp.Among 43 C.meningosepticum strains,26 strains (60.5%) produce MBL,but all strains (100%) produced extended-spectrum ?-lactamases (ESBLs);12 C.indologenes strains (68.8%) produced MBL;6 (60%) C.gleum strains had MBL.Genotypes of MBL in C.meningosepticum strains were Bla-B 1,2,3,5 and 11,and Bla-GOB 2,4,6 and 8,respectively.Only one genotype,namely CME-1,was identified for ESBL in C.meningosepticum.The genotype of MBL in 3 C.indolgenes strains was IND-1,and the 6 C.gleum strains contained CGB genotype.Meanwhile,there were 8 C.indolgenes strains and 3 C.gleum strains were confirmed to produce ?-lactamase,but their genotypes were unable to be detected using the current primers,implying that there were possible novel genotypes.Conclusions Investigation of genotypes distribution of ?-lactamase in Chryseobacterium/ Flavobacterium spp.can provide theoretical evidences and rational in the selection of antibiotics,control of noscomical infection and development of novel antibiotics.
3.Evaluation and comparison of MRSA 1D chromogenic agar medium and other 3 methods for detecting and identifying methicillin-resistant Staphylococcus aureus
Rong ZHANG ; Hong-Wei ZHOU ; Gong-Xiang CHEN ; Tie-Li ZHOU ;
Chinese Journal of Laboratory Medicine 2001;0(04):-
Objective To evaluate MRSA ID chromogenic agar medium for detecting and identifying methieillin-resistant Staphylococcus aureus(MRSA).Methods 118 Staphylococcus spp. clinical isolates(including 108 Staphylococcus and 10 coagulase-negative Staphylococcus(CNS)were identified by Bacteria Auto-Identification System.MRSA ID Chromogenic Agar,oxacillin screening agar, and cefoxitin disk diffusion method were compared for detecting methicillin-resistant Staphylococcus aureus (MRSA).Meanwhile,detection of mecA gene by polymerase chain reaction(PCR)was considered as the gold standard.Escheriehia coli ATCC25922,Staphylococcus aureus ATCC25923,Pseudomonas aeruginosa ATCC27853,Enterococcus faecalis ATCC29212 were used for interference test.Results meeA gene of 108 Staphylococcus was detected by PCR and 80 strains were positive.Compared with PCR,the agreement of screening MRSA by MRSA ID chromogenic agar,oxacillin screening agar and cefoxitin disk diffusion method were 100%,97.5%,and 100%,respectively.Only one methicillin-sensitive Staphylococcus(MSSA)and one CNS were shown in wrong color and became reseda micro-colonies on the MRSA ID chromogenic agar. Standard references strains did not grow on the MRSA ID chromogenie agar.Conclusion MRSA ID chromogenic agar can be used to rapidly and initiatively detect MRSA,which is useful to take quick infection control measures.
4.Psychological status of hospitalized patients of internal medicine and its related factors
Yueying XIANG ; Rong LIU ; Nasi HE ; Yuan GONG ; Xiaoping HUANG ; Jianneng HUANG ; Hong YIN
Chinese Journal of Health Management 2012;06(3):193-195
ObjectiveTo analyzing the psychological status of hospitalized patients of internal medicine and its impact factors.Methods Hospitalized patients of internal medicine underwent a survey by using Chinese Health Questionnaire (CHQ-12),Social Support Scale and Disease-Cognition Scale.Those with a score of >4 received further investigation of Symptom Check-List 90 (SCL-90).Correlation analysis was performed between all factors of SCL-90 and social support or disease-cognition scale score.Results There was no significant difference of psychological status between males and females ( P >0.05 ).All SCL90 factors were negatively correlated with social support,of which obsessive,paranoid,and phobic presented stronger negative correlations with social support and objective support (P < 0.05 ).Furthermore,all factors were negatively correlated with disease-cognition scale score.A significantly negative correlation between phobic factor and disease-cognition scale score was identified ( P < 0.05 ).Improvement was found in 26.2% patients after intervention.Conclusion Patients tend to show unhealthy emotion when they are under the stress of hospitalization.Hospitalization support system may need to be improved and patients' cognition of disease should be increased.
5.Expression of HCK Gene in Cardiomyocyte Differentiation of Mouse Embryonic Stem Cells
jie, GONG ; feng-rong, SUN ; ling-mei, QIAN ; xiang-qing, KONG ; yan-hui, SHENG ; rong, YANG ; ke-jiang, CAO
Journal of Applied Clinical Pediatrics 1986;0(01):-
Objective To explore the expression of HCK gene during the cardiomyocyte differentiation of mouse embryonic stem cells and analyze the role of HCK gene in maintenance of pluripotency of embryonic stem cells.Methods Mouse embryonic stem cells were cultured,then induced to differentiate into cardiomyocytes.Total RNAs were isolated from mouse embryonic stem cells in the differentiation days:0 day(D0),the second day(D2),the fourth day(D4),the sixth day(D6),the eighth day(D8),respectively.The levels of HCK mRNAs were assessed by the method of semi-quantitive reverse transcriptase-polymerase chain reaction(RT-PCR).In the meanwhile,Total proteins were also isolated from mouse embryonic stem cells in the differentiation D0,D2,D4,D6,D8,and the levels of HCK proteins were evaluated by Western-blot.Results HCK mRNAs could be detected in the mouse embryonic stem cells in D0 and D2,however,they were undetectable from D4 to D8.The expression of HCK mRNAs was rapidly down-regulated during cardiomyocyte differentiation of mouse embryonic stem cells.Expression of HCK proteins,which coincided with HCK mRNAs,down-regulated during differentiation and couldn't be detected in D4.Conclusions With the cardiomyocyte differentiation of mouse embryonic stem cells,the expression of HCK in the levels of mRNA and proteins are sharply down-regulated;HCK may play an important role in maintaining the pluripotency of embryonic stem cell.
6.Preliminary clinical application of contrast-enhanced MR angiography using three-dimensional timeresolved imaging of contrast kinetics
Chun-Shan YANG ; Song ZHANG ; Shi-Yuan LIU ; Xiang-Sheng XIAO ; Kang-Rong ZHOU ; Jin-Lin WANG ; Hui-Min LI ; Shan XIAO ; Wan-Qing GONG ;
Chinese Journal of Radiology 2001;0(03):-
Objective To assess the clinical application of contrast-enhanced MR angiography using three-dimensional(3D)time-resolved imaging of contrast kinetics(CE-MRA 3D-TRICKS).Methods TRICKS is a high temporal resolution(2—6s)MR angiographic technique using a short TR(2.8— 4.0 ms)and TE(0.9—1.3 ms),partial echo sampling and the central part of the k-space being updated more frequently than the peripheral part of the k-space.Pre-contrast mask 3D images are first acquired and 15--20 sequential 3D images following bolus injection of Gd-DTPA are then acquired.Results Thirty patients underwent contrast-enhanced MR angiography using TRICKS.Twelve vertebral arteries were well displayed on TRICKS.Seven of them showed normal,bilateral vertebral artery stenosis was shown in 1 case, and unilateral vertebral artery stenosis was shown in 4 wth aecompaning ipsilateral carotid artery bifurcation stenosis in one case.Bilateral renal artery showed normal in 4 cases,and the artery in transplanted kidney showed normal in one case and stenosis in another case.The cerebral artery showed normal in 2 cases, sagittal sinus thrombosis was detected in one case and intracranial arteriovenous malformation in one case. Pulmonary artery displayed normal in 3 cases,pulmonary artery thrombosis was seen in one case and pulmonary sequestration's abnormal feeding artery and draining vein was revealed in one case.The feeding artery in left lower limb fibrolipoma was showed in one case.The radial-ulnar artery artificial fistula stenosis was seen in one case,and left antebrachium hemangioma was showed in one case.Conclusion TRICKS can clearly delineate the whole body vascular system and can reveal any vascular abnormality.It is convenient and with high successful rate,which make it the first method of choice in displaying vascular abnormality.
7.Development and evaluation of immunoassay for zeranol in bovine urine.
Yuan LIU ; Cun-zhen ZHANG ; Xiang-yang YU ; Zhi-yong ZHANG ; Xiao ZHANG ; Rong-rong LIU ; Xian-jin LIU ; Zhen-ming GONG
Journal of Zhejiang University. Science. B 2007;8(12):900-905
A high affinity polyclonal antibody-based enzyme linked immunosorbent assay (ELISA) was developed for the quantification of zeranol in bovine urine. On the basis of urine matrix studies, the optimized dilution factors producing insignificant matrix interference were selected as 1:5 in pretreatment. In the improved ELISA, the linear response range was between 0.02 and 1 microg/ml, and the detection limit was 0.02 microg/ml for the assay. The overall recoveries and the coefficients of variation (CVs) were in the range of 82% to approximately 127% and 3.5% to approximately 8.8%, respectively. Thirty-six bovine urine samples spiked with zeranol (ranging from 0.2 to 10 microg/ml) were detected by the ELISA and liquid chromatography (LC) method, and good correlations were obtained between the two methods (R(2)=0.9643). We conclude that this improved ELISA is suitable tool for a mass zeranol screening and can be an alternative for the conventional LC method for zeranol in bovine urine.
Animals
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Calibration
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Cattle
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Chromatography, High Pressure Liquid
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Enzyme-Linked Immunosorbent Assay
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methods
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Indicator Dilution Techniques
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Zeranol
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immunology
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urine
8.Antimicrobial resistance and molecular epidemiological characteristics of clinical isolates of Staphylococcus aureus in Changsha area.
Ming-Xiang ZOU ; Rong-Rong ZHOU ; Wen-Jun WU ; Ning-Jie ZHANG ; Wen-En LIU ; Fu-Ping HU ; Xue-Gong FAN
Chinese Medical Journal 2012;125(13):2289-2294
BACKGROUNDIncreasing prevalence of Staphylococcus aureus (S. aureus), particularly methicillin-resistant S. aureus (MRSA) has been reported in China. In this study, we investigated the drug resistance characteristic, genetic background, and molecular epidemiological characteristic of S. aureus in Changsha.
METHODSBetween January 2006 and December 2008, 293 clinical isolates of S. aureus were collected from 11 hospitals in Changsha and identified by the Vitek-2 system. All the isolates were verified as MRSA by PCR amplification of both femA and mecA genes. K-B disk method was used to test drug sensitivity of S. aureus to antibiotics. Pulsed-field gel electrophoresis (PFGE) was performed for genotypic and homologous analysis of 115 isolates randomly selected from the original 293 clinical S. aureus isolates.
RESULTSS. aureus was highly resistant to penicillin, ampicillin, erythromycin, and clindamycin with resistant rates of 96.6%, 96.6%, 77.1%, and 67.2% respectively. All the isolates were susceptible to tecoplanin, vancomycin, and linezolid. MRSA accounted for 64.8% (190/293) of all the S. aureus strains. The 115 S. aureus isolates were clustered into 39 PFGE types by PFGE typing, with 13 predominant patterns (designated types A to M) accounting for 89 isolates. The most prevalent PFGE type was type A (n = 56, 48.7%) and 100.0% of type A strains were MRSA. PFGE type A included 13 subtypes, and the most prevalent subtype was subtype A1 (46.4%, 26/56). Strains with PFGE type A were isolated from eight hospitals (8/11), and both subtypes A1 and A4 strains were isolated in a university hospital.
CONCLUSIONSClinical isolates of S. aureus in Changsha were resistant to multiple traditional antibiotics. There was an outbreak of PFGE type A MRSA in this area and the A1 subtype was the predominant epidemic clone. Dissemination of the same clone was an important reason for the wide spread of MRSA.
Ampicillin ; pharmacology ; Anti-Bacterial Agents ; pharmacology ; China ; Clindamycin ; pharmacology ; Electrophoresis, Gel, Pulsed-Field ; Erythromycin ; pharmacology ; Humans ; Methicillin-Resistant Staphylococcus aureus ; drug effects ; genetics ; Microbial Sensitivity Tests ; Penicillins ; pharmacology ; Staphylococcus aureus ; drug effects ; genetics ; Vancomycin ; metabolism
9.Generation of a P19-alphaMHC-EGFP reporter line and cardiomyocyte differentiation.
Jie GONG ; Qi-jun ZHANG ; Jian WANG ; Feng-rong SUN ; Ling-mei QIAN ; Xiang-qing KONG ; Rong YANG ; Yan-hui SHENG ; Ke-jiang CAO
Chinese Journal of Cardiology 2008;36(8):691-694
OBJECTIVETo generate a P19-alphaMHC-EGFP reporter line and induce cardiomyocyte differentiation of this reporter line.
METHODSThe P19 cells were transfected with palphaMHC-EGFP, a P19-alphaMHC-EGFP reporter line was obtained after G418 selection and limited dilution of recombinant clones. The reporter line was induced to differentiate into cardiomyocytes which would beat and express green fluorescent protein. A comparison of cardiomyocyte differentiation rate and cTnI expression amount between the reporter line and the untransfected P19 cells was also performed. The ultrastructure was observed under transmission electron microscope.
RESULTSThe ultrastructure characteristics indicated cardiomyocytes-like changes on induction day 10. The beating cardiomyocytes which express GFP appear in the seventh induction day. The cardiomyocyte differentiation rate and cTnI expression amount of P19-alphaMHC-EGFP reporter line were similar as those in untransfected P19 cells (P > 0.05).
CONCLUSIONThe P19-alphaMHC-EGFP reporter line is of great benefit for identifying and purifying cardiomyocytes from undifferentiated P19 cells without influencing the differentiation of P19 cells. This feature makes P19-alphaMHC-EGFP reporter line a promising cell source for clinical cardiomyocyte replacement therapy.
Animals ; Cell Culture Techniques ; Cell Differentiation ; Cell Line ; Mice ; Myocytes, Cardiac ; cytology ; Stem Cells ; cytology ; Transfection
10.Preliminary study on the discrimination of putative periodontal pathogens with a metabonomics method.
Wei-xi LU ; Ya-fei WU ; Li-ying XIAO ; Ming-yun LI ; Qiang GUO ; Ping XIONG ; Xiang-ming JIA ; Xiao-rong XIAO ; Zhu ZHU ; Qi-mei GONG ; Wei LI
West China Journal of Stomatology 2009;27(3):310-316
OBJECTIVETo evaluate the feasibility of identifying oral pathogenic bacteria by comparing the metabolic profiling of putative periodontal pathogens and try to find a convenient and rapid way to discriminate oral microorganisms.
METHODSSuspensions of Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum with same density were prepared and cultured respectively at liquid BHI medium. Then the growth quantity was measured periodically through turbidimetry and the growth curves of the inoculated bacteria were completed. The culture solutions of stable growth phase were sampled and characterized by 1H-nuclear magnetic resonance 1H-NMR). The data of 1H-NMR spectroscope results were analyzed by principal components analysis (PCA).
RESULTSThe PCA showed the obvious clustering phenomena and the points of three groups differentially centralized to three clusters. Therefore, the NMR-based metabonomics profiles could discriminate the three different kinds of bacteria.
CONCLUSIONThe metabonomics is a potential classable method to identify the oral pathogenic bacteria.
Aggregatibacter actinomycetemcomitans ; Bacteria ; Fusobacterium nucleatum ; Metabolomics ; Mouth ; microbiology ; Porphyromonas gingivalis ; Prevotella intermedia