Table of Content

08 August 2012, Volume 27 Issue 8

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Study on the molecular mechanism of aminoglycoside resistance to Acinetobacter baumannii

2012, 27(8):  619-623. 

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Objective　To investigate the molecular epidemiology and mechanism of aminoglycoside resistance to Acinetobacter baumannii isolates. Methods　Agar-dilution was carried out to detect the minimum inhibitory concentration（MIC）, and enterobacterial repetitive intergenic consensus（ERIC）-polymerase chain reaction（PCR） was performed to analyze the molecular epidemiology of aminoglycoside-resistance isolates. Specific PCR，DNA sequencing，conjugation experiments were carried to confirm the transmission mechanism. Results　All the clinical isolates were resistant to most drugs including aminoglycosides, and ERIC-PCR showed the isolates belonged to 7 genotypes. Specific PCR and DNA sequencing revealed that all isolates encoded aminoglycoside-modifying enzyme genes, efflux pump gene and methylase gene. Conclusions　Producing of aminoglycoside-modifying enzyme and methylase mainly contribute to reduce the susceptibility of aminoglycosides in Acinetobacter baumannii. Efflux pump overexpression may as a cofactor in high-level aminoglycoside resistance. Vertical transmission and plasmid-mediated horizontal transmission are probably the principal epidemical mechanisms.

Changes of plasma concentrations of tryptophan，kynurenine and kynurenic acid in essential hypertensive patients

2012, 27(8):  624-627. 

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Objective　To investigate the correlations between the pathogenesis of essential hypertension （EH） with peripheral indoleamine 2，3-dioxygenase （IDO） and kynurenine aminotransferase （KAT） activities by measuring plasma concentrations of tryptophan （TRP），kynurenine （KYN） and kynurenic acid （KYNA）.   Methods　TRP，KYN and KYNA were measured by high performance liquid chromatography（HPLC） in the plasma of 100 patients with EH and 80 healthy controls. The enzyme activity was estimated by calculating the percentage of product/substrate，as IDO activity=KYN/TRP×100% and KAT activity=KYNA/KYN×100%. Results　TRP concentrations were （59.85±9.89）μmol/L，which were significantly higher in patients with EH than in healthy controls [（48.19±7.72）μmol/L，P＜0.001]. KYN concentrations were （2.01±0.48）μmol/L，which were significantly lower in patients with EH than in healthy controls [（2.17±0.43）μmol/L，P＜0.05]. The concentrations of KYNA in the patients with EH were （24.10±9.12）μmol/L，and were （23.59±7.27）μmol/L in healthy controls, and there were no statistical significances between the 2 groups（P>0.05）. The IDO activity was 3.40%±0.85%，which was significantly lower in the patients with EH than in healthy controls （4.54%±0.81%，P＜0.001）. KAT activity（1.20%±0.36%） in patients with EH was significantly higher than that in healthy controls （1.09%±0.27%，P＜0.05）. Plasma TRP concentration in the patients with EH had negative correlation with age （r=-0.316，P=0.001），and there was no correlation in the healthy controls（r=-0.208，P=0.064）. Plasma IDO activity in the patients with EH and the healthy controls had positive correlation with age （EH group：r=0.264，P=0.008；control group：r=0.305，P=0.006）. There was no obvious correlation with age between the 2 groups.  Conclusions　The increased concentrations of TRP and decreased concentrations of KYN in plasma of EH patients suggest that peripheral IDO activity in the TRP-KYN metabolism pathway may be involved in the pathogenesis of EH.

An epidemiological research on human papillomavirus infection among women in Sanya area

WANG Yufeng;LIN Ling;LIU Chunyu;LIU Bin;LI Zhixia;LI Qiang

2012, 27(8):  628-630. 

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Objective　To study the status of infection and the genotype distribution of human papillomavirus （HPV） among women in Sanya area. Methods　A total of 2 302 outpatient women in Sanya area from February 2009 to April 2011 were enrolled and determined for 21 HPV genotypes by DNA flow-through hybridization technique. Results　Among 2 302 cases, 562 cases were HPV positive, and the total positive rate was 24.41%. The highest infection rate （36.00%） was found in the age group of ≤20 years old, and the positive rate of HPV decreased with age. The total multiple infection rate was 58.54%. Among them, the double infection was common. The detection rate of detecting HPV genotypes were HPV16（21.06%）, HPV6（14.16%）, HPV58（13.02%）, HPV18（9.63%）, HPV52（9.29%）, HPV11（6.80%）, HPV31（6.34%）, HPVCP8304（5.21%）, HPV42（3.06%）, HPV53（2.04%）, HPV51（1.70%）, HPV66（1.70%）, HPV68（1.02%）, HPV33（0.91%）, HPV39（0.68%）, HPV44（0.68%）, HPV35（0.57%）, HPV45（0.57%）, HPV56（0.57%）, HPV43（0.57%） and HPV59（0.45%）. Conclusions　The prevalence of HPV infection and multiple infection are common in Sanya area. The most common high-risk genotypes are HPV 16, 58, 18, 52 and 31. The distributions have their own area character.

The clinical significance of Lunx mRNA detection in plasma and peripheral mononuclear cells in patients with lung cancer

ZHAO Wenjie;ZHOU Hongxing;WANG Sujian;WU Jiankang;WANG Jiaping;ZHANG Ping

2012, 27(8):  631-634. 

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Objective　To investigate the clinical auxiliary diagnosis significance of lung-specific X gene （Lunx） mRNA detection in plasma and peripheral mononuclear cells in patients with lung cancer. Methods　Lunx mRNA in plasma and peripheral mononuclear cells was detected by fluorescence quantitation polymerase chain reaction （PCR） in patients with lung cancer，benign lung disease，extrapulmonary tumor and healthy subjects.   Results　The positive rate of Lunx mRNA from plasma in lung cancer group was significantly higher than those in benign lung disease group （χ²=113.10，P＜0.01）， extrapulmonary tumor group （χ²=125.34，P＜0.01） and healthy subjects （χ²=100.33，P＜0.01）. The positive rate of Lunx mRNA in plasma in patients with Ⅲ-Ⅳ stages of lung cancer was significantly higher than that in patients with Ⅰ-Ⅱ stages of lung cancer （χ²=7.07，P＜0.05）. The positive rate of Lunx mRNA in peripheral mononuclear cells in lung cancer group was significantly higher than those in benign lung disease group （χ²=32.79，P＜0.01），extrapulmonary tumor group （χ²=44.44，P＜0.01） and healthy subjects （χ²=44.44，P＜0.01）. The positive rate of Lunx mRNA in peripheral mononuclear cells in patients with Ⅲ-Ⅳ stages of lung cancer was significantly higher than that in patients with Ⅰ-Ⅱ stages of lung cancer （χ²=24.52，P＜0.01）. For the auxiliary diagnosis of lung cancer, the sensitivity of Lunx mRNA detection in plasma was higher than that in mononuclear cells （χ²=36.46，P＜0.01）. The negative predictive value of the Lunx mRNA detection in plasma was higher than that in mononuclear cells （χ²=16.37，P＜0.01）.   Conclusions　The detection of Lunx mRNA in plasma and peripheral mononuclear cells can be used for the auxiliary diagnosis of lung cancer. The sensitivity in plasma is higher than that in mononuclear cells.

The inhibition of cell proliferation and invasion through miR-34a regulating YY1 in human renal carcinoma cell

WANG Mingli;LI Zhi;XU Jinyu;WENG Wenhao;XU Shanshan

2012, 27(8):  635-640. 

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Objective　To investigate miR-34a expression in renal carcinoma cell and the inhibitory effect and regulating mechanism of miR-34a on the proliferation and invasion in human renal carcinoma ACHN cells.  Methods　The expressions of miR-34a in cancer and pericancerous tissues were detected by real-time polymerase chain reaction（PCR）. ACHN cells were transfected with miR-34a mimics and there set blank control group, negative control group and miR-34a mimics group. The expression of miR-34a was measured by real-time PCR 24 h after transfection. The cell proliferation and cell cycle were determined by 5-dimethylthiazol-2-yl-2, 5-diphenyltetrazolium bromide （MTT） and flow cytometry. The invasive ability was examined by Transwell and Matrigel invasive assays. The mRNA and protein levels of YY1 were detected by real-time PCR and Western blot.   Results　The relative expression of miR-34a in cancer tissues （1.06±0.67） was significantly lower than that in pericancerous tissues （1.62±0.83, P＜0.01）. After transfection 24 h， an increase expression of miR-34a was noted in miR-34a mimics group （157.04±13.01） comparing with negative control group（P＜0.01）. Overexpression of miR-34a significantly inhibited the cell proliferation（P＜0.01）. The cell cycle was arrested at G0/G1 phases.Transwell and Matrigel invasive assays showed that the invasive ability of cells was significantly suppressed after transfection（P＜0.01）. YY1 gene had no mRNA expressions after transfection （P＞0.05）.  Conclusions　The expression of miR-34a is low in renal carcinoma，and it may be correlated with tumorigenesis，partially through regulating YY1.

Clinical application of the estimation of glomerular filtration rate in diabetes mellitus patients

LI Li;YANG Fan;MAO Kezi;LU Yide

2012, 27(8):  641-646. 

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Objective　To investigate the suitable equation of glomerular filtration rate（GFR） in estimating renal function for type 2 diabetes mellitus patients，and to compare the application effects of modification of diet in renal disease（MDRD）, reexpressed modification of diet in renal disease（rMDRD）, chronic kidney disease epidemiology collaboration（CKD-EPI） and cystatin C（Cys C）-based equations for GFR.  Methods　Clinical data from 178 type 2 diabetes mellitus patients were collected for the detection of serum creatinine（SCr） level，Cys C level and renal clearance of ^99mTc-diethylenetriamine pentaacetic acid-deoxyglucose（^99mTc-DTPA） which was used as the reference of isotopic GFR（iGFR）. According to the standard of the American Diabetes Association（ADA）, the 178 patients were classified into 3 groups [GFR：15-59, 60-89 and ≥90 mL/（min·1.73 m²）]. The equations for estimated GFR（eGFR） were compared with iGFR by paired t test，linear analysis，Bland and Altman procedures，receiver operating characteristic （ROC） curves and 15%，30% and 50% coincidences.  Results　When GFR<60 mL/（min·1.73 m²）， there was no significant difference between eGFR and iGFR in MDRD equation，but when GFR≥60mL/（min·1.73 m²），there was significant difference（P＜0.001）. There was no significant difference of eGFR for rMDRD equation in accuracy and diagnostic sensitivity. When GFR≥90 mL/（min·1.73 m²），the coincidences were higher in CKD-EPI equation than those in MDRD and rMDRD equation，but when GFR 60-89 mL/（min·1.73 m²），they were lower than those in Cys C GFR equation. The Cys C GFR equation had a better correlation with eGFR and iGFR，a less bias，a higher deviation and a higher coincidence than those in MDRD and rMDRD equations，especially when GFR≥90 mL/（min·1.73 m²）， only there was no significant difference between eGFR and iGFR, and Cys C GPR equation was better than the other 3 equations.  Conclusions　The 4 equations can estimate GFR accurately for type 2 diabetes mellitus patients when GFR<60 mL/（min·1.73 m²），but when GFR≥90 mL/（min·1.73 m²），Cys C GFR equation shows a significant superiority than the old SCr based equations.

The clinical significance on the combined detection of the serum BNP，hs-CRP，cTnI and UA in heart failure

HE Yan;LI Furong;DU Zongxiao;PIAO Wenhua

2012, 27(8):  647-650. 

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Objective　To investigate the clinical significance on the combined detection of B-type natriuretic peptide（BNP），high sensitive C reactive protein（hs-CRP），cardiac troponin I（cTnI）and uric acid（UA） in the diagnosis of heart failure（HF）. Methods　The serum levels of BNP，hs-CRP，cTnI and UA in 292 patients with different etiological factors and grades of HF[according to the cardiac function standard of New York Heart Association （NYHA）Ⅰ-Ⅳ] and 100 healthy controls were determined. The sensitivity and specificity of 4 parameters were evaluated by receiver operating characteristic （ROC） curve.  Results　The levels of BNP，hs-CRP，cTnI and UA in different grades of HF had statistical significance （P＜0.05）. In addition，the grade was worse，and its concentration was higher. The levels of BNP and hs-CRP showed statistical significance between HF NYHAⅠ grade patients and healthy controls （P＜0.05）, and the levels of cTnI and UA had no statistical significance between the healthy controls and HF NYHAⅠ grade patients（P＞0.05）. The sensitivity of combined detection was 90.1% in early HF diagnosis, which was higher significantly than that of the individual detection （P＜0.01）.  Conclusions　The significant clinical significance on the combined detection of the serum BNP，hs-CRP，cTnI and UA provides reference support in the diagnosis of early HF.

Influence of sample hemolysis on electrochemiluminescence immunoassay results

CHEN Haibin;LIANG Yebin;HUANG Huichang;ZHENG Muyang;MENG Xiujian

2012, 27(8):  651-653. 

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Objective　To study the influence of different degrees of sample hemolysis on electrochemiluminescence immunoassay（ECLIA） results. Methods　The blood samples were manually intervened into different degrees of hemolysis, and were used for detecting and analyzing the concentrations of 28 indices such as thyroid function, tumor markers, endocrine hormones and so on in various groups of samples. Results　Sample hemolysis can reduce the concentration of insulin（INS），and the concentration of folic acid （FA） and neuron-specific enolase （NSE） were increased. Severe hemolysis can make the results to have a higher trend on determination of ferritin，however, it was not statistically significant, and the determination results of other indices was not affected by hemolysis. Conclusions　The results of most hemolytic samples except certain indices are not affected. The determinations of INS, FA and NSE should absolutely avoid hemolytic samples to ensure the accuracy of results.

Research on the significance of 1-palmitoyl-sn-glycero-3-phosphocholine in the diagnosis of ischemic heart disease

XU Weijia;WEN Jie;LIU Yi

2012, 27(8):  654-658. 

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Objective　To screen the most significantly changed metabolite in the sera of patients suffering from ischemic heart disease（IHD）, and investigate its clinical significance. Methods　A total of 10 biochemical parameters involved in the diagnosis of ischemic diseases or infarction were determined including alanine aminotransferase（ALT），aspartate aminotransferase（AST），gamma-glutamyltransferase（GGT），alkaline phosphatase（ALP），lactate dehydrogenase（LDH），alpha-hydroxybutyrate dehydrogenase（α-HBDH），creatine kinase-MB（CK-MB），creatine kinase（CK），C reactive protein（CRP） and cardiac troponin（cTnT） between 52 IHD patients and 45 controls. A strategy of high performance liquid chromatography-mass spectrometry（HPLC-MS） technique was used to explore the major metabolic changes in the sera of the 2 groups in view of metabolomics. The acquired data were subjected to the significance analysis to select and identify the most importantly changed metabolite. Results　The results showed that 4 ion fragments were found between the 2 groups，whose average mass charge ratios （m/z） were 496.33，478.34，184.13 and 991.71. They were finally confirmed to be originated from 1-palmitoyl-sn-glycero-3-phosphocholine（C16：0）. The concentration of this metabolite was （76.35±18.28）μmol/L in IHD patients，which was higher than that in the controls [（64.24±15.56）μmol/L，P＜0.001]. The 9 traditional serum parameters were not different between the patients and the controls（P＞0.05）, except the slightly elevated AST in the IHD group （P＜0.05）. Receiver operating characteristic（ROC） curve indicated that area under the curve of 1-palmitoyl-sn-glycero-3-phosphocholine（C16：0）（0.90±0.03） was larger than that of the other traditional biochemical parameters（P＜0.05）. Conclusions　1-palmitoyl-sn-glycero-3-phosphocholine（C16：0） has clinical significance for the diagnosis of IHD.

Study on the expression and significance of Toll-like receptor 2 and 4 of peripheral blood mononuclear cells in patients with early-stage rheumatoid arthritis

QIAN Lei;WANG Xiaoying;Lü Lijun;CHEN Dexun

2012, 27(8):  659-662. 

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Objective　To investigate the effects of Toll-like receptor（TLR）2 and TLR4 of peripheral blood mononuclear cells（PBMC） in patients with early-stage rheumatoid arthritis （RA） and the responsiveness of TLR4 to lipopolysaccharide（LPS） and TLR2 to biglycan（BGN）.  Methods　The expressions of TLR4⁺/CD14⁺ monocytes，TLR2 mRNA, TLR4 mRNA and cytokines of interleukin-6（IL-6） and tumor necrosis factor alpha（TNFα） in supernatants were analyzed by flow cytometry，real-time fluorescence quantitation reverse transcription（RT）- polymerase chain reaction（PCR） and enzyme-linked immunosorbent assay（ELISA） respectively in PBMC before and after stimulation.  Results　The TLR2 mRNA in PBMC of RA patients were significantly higher than those of healthy controls（P＜0.01）. The TLR4 mRNA of RA patients were significantly lower than those of healthy control. LPS increased TLR4 mRNA of RA patients（3.50-fold） and decreased TLR4 mRNA of healthy controls（0.11-fold）. LPS and BGN increased cytokines′ production in all groups，while the increased folds of IL-6 and TNFα in PBMC from RA patients were significantly higher than the cells from healthy controls.  Conclusions　TLR2 and TLR4 of PBMC display important roles in the development of early-stage RA.

Influence of different bacteria or fungi on the determination of urinary sediments by UF-1000i urinary sediment analyzer

ZHENG Qin;ZHAO Ru;HUANG Qian;XIONG Ming;SU Jun

2012, 27(8):  663-666. 

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Objective　To study the influence of different bacteria or fungi with different levels on the determination of urinary red blood cell（RBC） count by UF-1000i urinary sediment analyzer（UF-1000i），so as to provide more reliable laboratory evidence in the diagnosis and follow-up of urinary tract diseases.  Methods　The urine RBC counts were analyzed by UF-1000i from urine mixture specimens and hematuria specimens with Blastomyces albicans，in parallel with manual microscopy respectively.  Results　The Gram-negative bacilli（Escherichia coli，Klebsiella pneumonia and Pseudomonas aeruginosa） and Gram-positive cocci （Staphylococcus aureus） isolated from urine specimens of urinary tract infection and low-concentration Blastomyces albicans（<106 mL） were not impact the urine RBC count detected by UF-1000i. High concentration bacteriuria specimens（=109/mL） of Enterococcus faecalis caused the false-positive RBC count. The urine RBC count determined by UF-1000i was caused false-positive by Candida albicans, when the concentration of fungi was more than 106/mL. The concentration of Candida albicans reached to 108/mL，and the hematuria specimen RBC count would falsely increased（P=0.006）.  Conclusions　UF-1000i has special bacterial examination channel, and it could prevent the interference with urine RBC count obviously. However, it is hard to prevent the interference of high concentration fungi，which would cause false-positive urine RBC count. It is recommended to correct the RBC count by manual microscopy in order to provide accurate laboratory results.

Evaluation on nucleated red blood cell detection by XE-5000 automated hematology analyzer

MU Yueyi;LI Xiaomei;LI Yong;XIA Yonghui;TIAN Peng;YANG Jiayu;ZHANG Huamei;CAO Zeng

2012, 27(8):  667-670. 

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Objective　To evaluate the clinical application significance and methodology characteristic of Sysmex XE-5000 automated hematology analyzer （XE-5000） to count nucleated red blood cell（NRBC）.  Methods　The repeatability，linear range and carryover rate of XE-5000 were evaluated. A total of 391 blood samples were measured, and the alarm and quantitative determination of XE-5000 for NRBC were evaluated. The results were compared with manual microscopy. Results　The XE-5000 results showed good repeatability, linear range and low carryover rate. When NRBC％＞5.0％，it caused false increasing of leucocyte and lymphocyte. The results of manual microscopy were as a gold standard. The correlation was good between the XE-5000 and manual microscopy（r=0.997）. The sensitivity for NRBC was 100.0％，and the specificity was 85.4％.  Conclusions　NRBC counting by XE-5000 is sensitive，accurate and reliable，which is suitable for the clinical detection of NRBC in peripheral blood.

Performance analysis on colloidal gold immunoassay for fecal occult blood test kit and its primary evaluation of clinical application

YANG Lin;LU Renquan;GUO Lin

2012, 27(8):  671-673. 

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Objective　To evaluate the performance and clinical application of the fecal occult blood test kit by colloidal gold immunoassay.   Methods　A total of 150 stool samples were collected randomly from inpatients. The comparative determination of these samples were performed by experimental kit（bioMerieux Co., Ltd.） and control kit（Shanghai Chemtron Biotech Co., Ltd.）. The results were statistically analyzed. The primary clinical application was evaluated.   Results　The experimental kit had a good sensitivity （40 ng/mL）, and its specificity and repeatability met the needs of clinical application. The determination results suggested that it could be consistent with control kit results and clinical pathogenetic condition.   Conclusions　The fecal occult blood test kit by colloidal gold immunoassay is simple, reliable, and suitable for clinical application.