Table of Content

30 October 2013, Volume 28 Issue 10

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Orignal Article

The distribution and antibiotic resistance of nosocomial infection pathogens in cancer patients after radiotherapy and chemotherapy

ZHANG Jing,LU Renquan,LING Yun,GUO Lin.

2013, 28(10):  873-877.  DOI: 10.3969/j.issn.1673-8640.2013.10.002

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Objective To investigate the pathogen infection and antibiotic resistance in patients with malignant tumor undergoing radiotherapy and chemotherapy in Shanghai Cancer Center, Fudan University during recent 2 years. Methods A total of 305 strains of pathogens were identified from clinical specimens in malignant tumor patients undergoing radiotherapy and chemotherapy by VITEK-32 automatic microbiology analysis system and common methods from January 2010 to December 2011. The drug sensitivity test was detected according to the guideline of the Clinical and Laboratory Standards Institute (CLSI). Results The most common specimen sources were respiratory tract,followed by urine and blood. The main pathogens were gram negative organisms (57.7% and 46.5%),in which still Acinetobacter baumannii,Escherichia coli and Pseudomonas aeruginosa were the first 3 main kinds of pathogens during the 2 years. The prevalences of extended spectrum beta-lactamases (ESBLs) in the strains of Escherichia coli were 57.1% in 2010 and 28.6% in 2011. The following infection was fungus infection (27.0% in 2011 and 20.8% in 2010),among which Candida was the main pathogen. The gram positive organisms in 2011 were higher than those in 2010,and the percentage of methicillin-resistant Staphylococcus aureus decreased from 32.0% to 21.3% of the gram positive organisms. No strains were found resistant to vancomycin. Conclusions The main pathogens are contributed mostly by gram negative organisms in patients with malignant tumor undergoing radiotherapy and chemotherapy. Attentions should be paid to the pathogen infection determination and antibiotic resistance monitoring. It will provide the reference for the clinical dignosis.

Preliminary study on drug resistance and mutant prevention concentration of Klebsiella pneumoniae

SUN Jie, XU Weixin, SHI Jianying, JI Jian, LIU Chunhong, CAO Wenjun.

2013, 28(10):  878-881.  DOI: 10.3969/j.issn.1673-8640.2013.10.003

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Objective To study the drug resistance of Klebsiella pneumoniae to common antibiotics, and to measure the mutant prevention concentration (MPC) of fluoroquinolone (FQNs) and analyze the anti-drug resistance mutation ability. Methods The minimum inhibition concentration (MIC) was determined by micro-dilution method, and MPC was determined by agar dilution method. The drug resistance to common antibiotics was measured by K-B method. Results A total of 30 clinical isolates of Klebsiella pneumoniae had extended-spectrum beta lactamases (ESBLs) positive (15 isolates, positive group) and negative (15 isolates, negative group). The results showed that the drug resistance to common antibiotics had significant difference. The drug resistance rate of ESBLs positive group was higher than that of ESBLs negative group.The MPC, MIC anddrug resistance selection index (SI) of levofloxacin and moxifloxacin for ESBLs positive and ESBLs negative Klebsiella pneumoniae showed different distribution characteristics. Conclusions For the 30 isolates of Klebsiella pneumoniae, the drug resistance to common antibiotics of ESBLs positive isolates is serious, and MPC and SI are significantly higher than those of ESBLs negative isolates.

Correlationship of serum alpha₁-antitrypsin and C reactive protein levels with chronic obstructive pulmonary disease

PANG Guoju,LIU Huaiping.

2013, 28(10):  882-884.  DOI: 10.3969/j.issn.1673-8640.2013.10.004

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Objective To investigate serum C reactive protein(CRP) and alpha₁-antitrypsin(α₁-AT) levels in patients with chronic obstructive pulmonary disease(COPD) and the correlation with the severity of COPD. Methods Serum levels of CRP and α₁-AT were determined by immunoturbidimetry in 43 patients with acute exacerbation chronic obstructive pulmonary disease(AECOPD),47 patients with stable COPD and 40 healthy subjects (healthy control group),and the results were analyzed statistically. Results Serum levels of CRP and α₁-AT had emerge rise trend with the severity of disease. Serum levels of CRP and α₁-AT in AECOPD group and stable COPD group were higher than those in healthy control group(P<0.01) . Serum levels of CRP and α₁-AT in AECOPD group were higher than those in stable COPD group(P<0.05). Serum level of CRP was correlated positively with serum level of α₁-AT in AECOPD group (r=0.704,P<0.01). However, serum level of CRP was uncorrelated with serum level of α₁-AT in stable COPD group (r=0.232,P>0.05). Conclusions The airway inflammation is continuously existing in patients with COPD. CRP and α₁-AT are sensitive indicators for the patients with COPD, and are positively correlated with the severity of airway inflammation. CRP is more sensitive than α₁-AT for AECOPD patients, and there is no obvious increasing of α₁-AT level.

The relationship of plasma Lp-PLA2 level with the stability of carotid artery atherosclerotic plaque and the severity of neurological impairment in patients with acute ischemic stroke

LONG Lu,WANG Zhongming,CHEN Zhen,TAO Ya,WANG Kun, YI Bin.

2013, 28(10):  885-889.  DOI: 10.3969/j.issn.1673-8640.2013.10.005

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Objective To investigate the relationship of plasma lipoprotein-associated phospholipase A2(Lp-PLA2) level with the stability of carotid artery atherosclerotic plaque and the severity of neurological impairment in patients with acute ischemic stroke. Methods According to the results of cervical vascular color Doppler ultrasound examination,106 patients with acute ischemic stroke were classified into unstable plaque group(44 cases),stable plaque group(38 cases)and non-plaque group(24 cases). A total of 40 healthy subjects were enrolled as healthy controls. Plasma Lp-PLA2 levels were determined by enzyme-linked immunosorbent assay (ELISA),and the severity of neurological impairment were assessed by the National Institute of Health Stroke Scale(NIHSS) score. Results Compared with the healthy controls,the level of plasma Lp-PLA2 was significantly higher in patients with acute ischemic stroke (Z=-6.995,P<0.05). Plasma levels of Lp-PLA2 in stable plaque group and unstable plaque group were significantly higher than those in non-plaque group with statistical significance (Z=-5.670,P<0.05). Plasma levels of Lp-PLA2 in unstable plaque group were significantly higher than those in stable plaque group with statistical significance (Z=-6.185,P<0.05). According to the results of receiver operating characteristic (ROC) curve,the best cut-off value of Lp-PLA2 was 137.00 μg/L. The sensitivity was 81.8%, and the specificity was 95.1%. The severity of neurological impairment increased with the levels of plasma Lp- PLA2(χ²=74.233,P<0.05). The NIHSS score was positively correlated with the levels of plasma Lp-PLA2(r_s=0.861,P<0.05). Conclusions The level of plasma Lp-PLA2 is related to the stability of carotid artery atherosclerotic plaque and the severity of neurological impairment in patients with acute ischemic stroke. Plasma Lp-PLA2 can serve as a predictive indicator of carotid artery atherosclerotic plaque stability.Meanwhile,it plays an important role in monitoring the conditions of patients with acute ischemic stroke.

Significance of activin A in the diagnosis of suspected ectopic pregnancy

YANG Xiaoning,HU Hua,FU Xiufang,GU Jiashi.

2013, 28(10):  890-892.  DOI: 10.3969/j.issn.1673-8640.2013.10.006

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Objective To investigate the significance of activin A in the early diagnosis of ectopic pregnancy(EP). Methods A total of 109 patients with suspected EP were classified into 2 groups according to the follow-up results:62 patients with EP and 47 patients with intrauterine pregnancy (IUP). The serum levels of activin A and beta-human chorionic gonadotropin (β-HCG) were determined in 109 patients with suspected EP by enzyme-linked immunosorbent assay (ELISA),and the receiver operating characteristic(ROC) curves were established. The diagnosis significance of activin A and β-HCG for EP was analyzed. Results The levels of activin A and β-HCG in EP group were significantly lower than those in IUP group(P<0.05). The area under ROC curves of activin A (0.827) was higher than that of β-HCG(0.710). In activin A＜260 pg/mL for the diagnosis of EP,the sensitivity and specificity were 85.5% and 72.3%, respectively. Conclusions The determination of serum activin A can be considered as an early diagnosis parameter of EP.

Clinical significance on the determinations of IL-17 and IL-23 from bronchoalveolar lavage fluid in patients with chronic obstructive pulmonary disease

MOU Jiaoyue,YE Tingjun,FAN Qishi.

2013, 28(10):  893-896.  DOI: 10.3969/j.issn.1673-8640.2013.10.007

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Objective To determine the changes of interleukin 17 (IL-17) and interleukin 23 (IL-23) from bronchoalveolar lavage fluid (BALF) in patients with chronic obstructive pulmonary disease(COPD), and to provide the reference for diagnosing and understanding the inflammation development of COPD. Methods On the basis of cytological classification for BALF, clinical data and forced expiratory volume in first second to forced vital capacity ratio(FEV1%), acute exacerbation COPD group, stable phase COPD group and non-COPD control group were determined, respectively. The IL-17 and IL-23 levels were determined by enzyme-linked immunosorbent assay (ELISA). Results The IL-17 and IL-23 levels and FEV1% increased more significantly in acute exacerbation COPD group than in stable phase COPD group and control group (P<0.01) . There was a positive linear correlation between BALF IL-17 and IL-23 in COPD groups (r²=0.698 5,P<0.001), however, there was a negative linear correlation of BALF IL-17 and IL-23 with FEV1% in COPD groups (r²=-0.527 9 and -0.541 0, P<0.001). Conclusions The IL-17 and IL-23 levels with local lung inflammatory have correlations with the changes of COPD and the clinical manifestations,especially for IL-17,which may lead to persistent inflammation. Overall, the determinations of IL-17 and IL-23 are useful for the diagnosis and surveillance of COPD.

Research on the correlation of the acetaldehyde dehydrogenase ALDH7A1 gene poly morphism and type 2 diabetes mellitus

LIU Guozheng, LIU Fangfang, YAN Wenqiang, LIU Shuang, XIE Mingshui, XU Chengkai, JIANG Congqing.

2013, 28(10):  897-900.  DOI: 10.3969/j.issn.1673-8640.2013.10.008

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Objective To study the correlation of ALDH7A1 gene polymorphism locus rs2306619, rs2306618, rs12514417, rs2306617 and rs4379190 with type 2 diabetes mellitus among Chinese Hubei Suizhou Han population. Methods A total of 220 healthy controls and 240 unrelated patients with type 2 diabetes mellitus were enrolled, and all of them were from Han urban residents in Hubei Suizhou. The DNA was extracted from white blood cell of peripheral blood by standard method. The 5 polymorphism loci (rs2306619, rs2306618, rs12514417, rs2306617 and rs4379190) were amplified through designed sequence primer, and the sequence analysis was performed. The single nucleotide polymorphism and linkage disequilibrium of ALDH7A1 gene 5 polymorphism loci were analyzed statistically by SHEsis software. The differences and haplotypes of 5 locus allele frequencies and genotype frequencies were compared between type 2 diabetes mellitus group and control group. Results Five locus minimum allele frequencies were 0.492, 0.084, 0.124, 0.277 and 0.259. Linkage disequilibrium test showed 5 loci with strong linkage disequilibrium. The ALDH7A1 gene 5 locus allele frequencies and genotype frequencies in the 2 groups had no statistical significance. Haplotype analysis showed C-A-A-A-T, T- A- A- A-T, T-A-A-G-C, T-A-C-A-T and T-G-A-G-C haplotypes, and the 5 haplotypes intype 2 diabetes mellitus group and control group had no statistical significance. Conclusions The 5 polymorphism loci of ALDH7A1 gene may be the susceptibility genes for the patients with type 2 diabetes mellitus in Chinese Hubei Suizhou Han population. The results of the present study are needed to validate in larger populations.

Clinical significance of peripheral blood CD3⁺, CD4⁺ and CD8⁺ T lymphocyte subset determination in patients with tumor

YAN Jian, YUAN Yongming, ZHANG Shu, PENG Xiancong

2013, 28(10):  901-903.  DOI: 10.3969/j.issn.1673-8640.2013.10.009

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Objective To investigate the changes of peripheral blood T lymphocyte subsets (CD3⁺, CD4⁺, CD8⁺ and CD4⁺/CD8⁺) in patients with tumor and the clinical significance. Methods A total of 100 patients with tumor (including 40 patients with lung cancer, 40 patients with gastrointestinal cancer and 20 patients with other tumors) and 90 healthy controls were enrolled. Peripheral blood T lymphocyte subsets′ absolute counts and immunoglobulin (IgG, IgA and IgM) were determined. The differences between the 2 groups were analyzed statistically. Results T lymphocyte subsets′ absolute counts were CD3⁺ [(1 041±358)/μL], CD4⁺[(689±263)/μL], CD4⁺/CD8⁺ ratio (1.15±0.40) in tumor group, which were significantly lower than those in control group (P<0.05), and CD8⁺[(602±223)/μL] increased slightly compared with that in control group (P>0.05). IgG, IgA and IgM were higher in tumor group than in control group. Conclusions Tumor patients have immune dysfunction, and the changes of T lymphocyte subsets′ counts have significance on the monitoring and prognosis in patients with tumor.

Separation and purification of HbA_1c and HbA₀

TU Guohua,WANG Bijun,ZHU Yunjiao,QIN Ying,JIANG Xugan.

2013, 28(10):  904-907.  DOI: 10.3969/j.issn.1673-8640.2013.10.010

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Objective To separate and purify hemoglobin A_1c(HbA_1c) and hemoglobin A₀(HbA₀) from red blood cells of healthy subjects. Methods Collecting whole blood from healthy subjects, the red blood cells were separated and prepared for hemolysis. HbA_1c and HbA₀ were separated by weak acid positive ion exchange chromatography and purified by boric acid agarose affinity chromatography respectively. Results Hand-loaded column with weak acid positive ion exchange chromatography was used to separate and collect the peaks of HbA_1c and HbA₀ from healthy subjects′ red blood cell hemolysis, and the purities were 79.78% and 97.42% by high performance liquid chromatography (HPLC) respectively. After being purified by boric acid agarose affinity chromatography,the final purities of HbA_1c and HbA₀ were 94.3% and 99.7% respectively. Conclusions The single ion exchange chromatography for separating the glycosylated hemoglobin is not ideal,and it must be combined with affinity chromatography to obtain a high purities of HbA_1c and HbA₀.

Study on the determination of pregnancy associated plasma protein A by ELISA

JIN Baoying,ZENG Zhaowei,LI Huiqiang.

2013, 28(10):  908-912.  DOI: 10.3969/j.issn.1673-8640.2013.10.011

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Objective To establish a enzyme-linked immunosorbent assay (ELISA) for the diagnosis method of pregnancy associated plasma protein(PAPP-A). Methods Sera from full-term pregnant women were purified by molecular sieve and ion exchange chromatography,and then the biotin labeling PAPP-A and enzyme-labelled PAPP-A antibody were prepared. Traditional ELISA was used to detect the samples,and the sensitivity,specificity,recovery and stability were observed. The results of 30 samples from first trimester pregnant women were kit from DRG of USA. Results The result of the purification by molecular sieve and ion exchange chromatography was good. The range of standard curve was 1-50 μg/mL. The sensitivity was 0.35 μg/mL,the recovery was>100%,and the stability and specificity were good. The method had good correlation with the kit from DRG of USA,and the regression equation was Y=2.286 1+1.026 9X(r=0.982, P<0.01). Conclusions Biotin labeling PAPP-A is made, and the connection ability of biotin and avidin can be reflected in the ELISA. The ELISA is meaningful in the determination of PAPP-A.

Establishment of double antibody sandwich ELISA for the determination of human ubiquitous mitochondrial creatine kinase

ZHANG Jian,JIAN Minhua,JIANG Lingli,GE Danhong.

2013, 28(10):  913-916.  DOI: 10.3969/j.issn.1673-8640.2013.10.012

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Objective To establish double antibody sandwich enzyme-linked immunosorbent assay (ELISA) for the determination of ubiquitous mitochondrial creatine kinase(uMtCK) in human serum. Methods Seven strains of monoclonal antibodies against human uMtCK were prepared with prokaryotic expression recombinant proteins from rats, and the best combination of double antibodies was selected by checkerboard titration method. Double antibody sandwich ELISA for the determination of human uMtCK was established by using HRP-labeled McAb 19F11 as a detection antibody and McAb 3C1 as a capture antibody. The methodology performance was evaluated. The serum concentrations of uMtCK in 50 healthy subjects and 85 patients with positive tumor markers [alpha fetal protein (AFP) and carcinoembryonic antigen (CEA)] were determined. Results The double antibody sandwich ELISA for the determination of human uMtCK was established by selecting the combination of double antibodies. The detection limit was 28.8ng/mL, and the within-run and between-run coefficients of variation (CV) were 4.5%-7.7％ and 8.2%-13. 5％, respectively.The average recovery was 97.1％. The serum concentrations of uMtCK in 50 healthy subjects were negative. The 15 of 85 patients with positive tumor markers had positive results of uMtCK, and the concentration was 52.8-1 442.2 ng/mL. Conclusions The established double antibody sandwich ELISA can be used to determine uMtCK in human serum.

Comparison on the performance verification of automated hematology analyzer Sysmex XE-2100,Beckman-Couler LH750 and MINDRAY BC-5800

QIAO Rui,CUI Yanmei,LI Jing,PENG Jinhong,YANG Shuo,WANG Hongya,ZHANG Jie.

2013, 28(10):  917-920.  DOI: 10.3969/j.issn.1673-8640.2013.10.013

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Objective To verify the performance of automated hematology analyzers in clinical laboratories, and provide the reference for choosing the referred analyzer of a clinical laboratory. Methods A total of 15 fresh whole blood specimens assigned values and being provided by the National Center for Clinical Laboratory were used to evaluate trueness, 3 levels of quality control materials were used to evaluate intra-day and inter-day precisions, 5 dilutions were assayed to verify linearity, and the score of each analyzer was adopted to determine the referred analyzer. Results Correalation coefficient R values of white blood cell (WBC), red blood cell (RBC), hemoglobin (Hb), hematocrit (HCT) and platelet(PLT) of each analyzer were 0.953-0.998. However,R values of RBC mean corpuscular volume (MCV) of each analyzer were 0.751-0.821. Except PLT of XE-2100,the other parameters of each analyzer showed that the 95% confidence intervals of slope included the value 1, and the 95% confidence intervals of Y-intercepts included the value 0. Except PLT inter-day precision of BC-5800, the intra-day and inter-day precisions of the other analyzers were complied with the specification of each analyzer. Analytical measuring intervals of WBC and PLT of LH750 and BC-5800 were wider than those of XE-2100. Sorted from high to low,the rates of general performance of each analyzer were LH750,BC-5800 and XE-2100 in sequence. Conclusions The performance verification of XE-2100, LH750 and BC-5800 shows roughly satisfactory results. Meanwhile,LH750 is the most qualified as the referred hematology analyzer in the clinical laboratory.

Application evaluation of UF-1000i urine formed element analyzer in the screening of urinary tract infection

DING Zhixiang,ZHANG Hong,JIN Wentao,QIAN Gaochao.

2013, 28(10):  921-924.  DOI: 10.3969/j.issn.1673-8640.2013.10.014

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Objective To evaluate the application significance of UF-1000i urine formed element analyzer in the screening of urinary tract infection(UTI). Methods Medistream urine samples were collected from 148 patients with suspected UTI. Bacterium (BACT) counts and white blood cell (WBC) counts were determined by UF-1000i urine formed element analyzer. By comparing with quantitative urine culture as the golden standard of UTI,the positive cut-off criteria for UF-1000i was set through receiver operating characteristic (ROC) curve. The sensitivity,specificity,positive predictive value,negative predictive value and accuracy were evaluated. Results The cut-off values for BACT and WBC counts were 325/μL and 48/μL,respectively. The sensitivity,specificity,positive predictive value,negative predictive value and accuracy of BACT and WBC counts were 78.3%,92.2%,81.8%,90.4%,87.8% and 73.9%,81.4%,64.2%,87.4%,79.1%, respectively. The combined determination of BACT and WBC counts by UF-1000i showed that the sensitivity, specificity, positive and negative predictive values and accuracy were 95.7%,70.6%,59.5%,97.3% and 78.4%, respectively. Conclusions UF-1000i urine formed element analyzer may be a screening tool for UTI.

Performance evaluation of TOSOH HLC-723 G8 automatic glycohemoglobin analyzer

CHEN Jian

2013, 28(10):  925-927.  DOI: 10.3969/j.issn.1673-8640.2013.10.015

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Objective　To evaluate the performance of TOSOH HLC-723 G8 automatic glycohemoglobin analyzer (HLC-723 G8) for the determination of glycosylated hemoglobin A_1c (HbA_1c) with the principle of ion-exchange high-performance liquid chromatography (HPLC). Methods　The precision,correlation,linearity and carryover of HLC-723 G8 were evaluated. The reference interval of HbA_1c was verified among 54 healthy subjects. Results　The within-run and between-run coefficients of variation (CV) of both low and high levels of HbA_1c were 0.00%,0.47% and 0.76%, 0.31%, respectively. HLC-723 G8 and HLC-723 G7 for the determination of HbA_1c showed good correlation (r²=0.999,P<0.001). The expected bias was 0.06,the relative bias was 0.83%,and the bias coincidence was 100%. An excellent linearity was found for HbA_1c in the range of 5%-12% (r²=0.999 6,P<0.001).There was no obvious carryover in both low and high level samples, and the carryover rate was 1.12%. The levels of HbA_1c among the 54 healthy subjects were 4.7%-6.3% in the range provided by manufacturers. Conclusions　HLC-723 G8 has a good performance, and can be used in clinical application.