Table of Content

20 October 2016, Volume 31 Issue 10

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Orginal Article

Liquid biopsy for monitoring EGFR-TKI drug resistance in patients with non-small-cell lung cancer

LIN Cheng, JIANG Tang

2016, 31(10):  835-843.  DOI: 10.3969/j.issn.1673-8640.2016.010.001

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As a milestone in the treatment of non-small-cell lung cancer,epidermal growth factor receptor （EGFR）-tyrosine kinase inhibitor（TKI） drugs significantly prolong the lifetime of patients with non-small-cell lung cancer and improve the patients' life quality. The frequency of EGFR mutation among non-small-cell lung cancer patients in Asia is higher than that among white men,so Asian patients have more benefits from EGFR-TKI drugs. However,one of the most important problems in targeted drugs is drug resistance. All EGFR-mutated patients who initially benefit from EGFR-TKI drugs develop progressive disease eventually,usually after approximately 1 year since treatment. Nowadays,due to drug resistance,there have been 3 generations of EGFR-TKI drugs. For the mechanisms of EGFR-TKI drug resistance,point mutation EGFR T790M and C797S are of special concern. Dynamic monitoring of drug resistance must be performed in patients who use EGFR-TKI drugs to achieve better therapeutic effect. Pathological biopsy is a gold standard for the diagnosis of cancer. However,pathological biopsy can not be competent for the dynamic monitoring of non-small-cell lung cancer patients,due to the risk of operation,the difficulty of getting tissue samples,being unable to determine in real-time,tumor heterogeneity and so on. Liquid biopsy is a new,non-invasive,convenient and highly reliable determination method. Liquid biopsy shows great potential on early diagnosis and screening,evaluation of therapeutic effect and prognosis and monitoring drug resistance by determing circulating tumor cell（CTC）,circulating tumor DNA（ctDNA）and tumor-released exosome in blood. This article reviews the progress of EGFR-TKI drugs and liquid biopsy,and assesses the potential of liquid biopsy in EGFR-TKI drug resistance of patients with non-small-lung cancer.

Molecular characteristics and drug resistance of Staphylococcus aureus isolated from patients with skin and soft tissue infections in Haian

CHEN Ye, DONG Yefeng, DONG Deping

2016, 31(10):  844-847.  DOI: 10.3969/j.issn.1673-8640.2016.010.002

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Objective To study the molecular characteristics and drug resistance of Staphylococcus aureus isolated from patients with skin and soft tissue infections（SSTI） at Nantong University Haian Hospital in 2014. Methods A total of 26 isolates of Staphylococcus aureus were collected from patients with SSTI at Nantong University Haian Hospital,and the isolates were identified by phenotypic tests. Drug susceptibility test was performed by disk diffusion method. Panton-Valentine leukocidin（pvl） gene was determined by polymerase chain reaction（PCR）. Staphylococcal cassette chromosome mec（SCCmec） typing was performed by multi-PCR,and Staphylococcus protein A（spa） gene typing and multi-locus sequence typing（MLST） were performed by PCR and sequencing.Results A total of 5 isolates of methicillin-resistant Staphylococcus aureus（MRSA） were determined. Among them,3 isolates were ST630-t4549-SCCmec-Ⅴ,1 isolate was ST59-t437-SCCmec-Ⅰ,and 1 isolate was ST5-t311-SCCmec-Ⅰ. There was no resistant isolate to vancomycin,teicoplanin,sulfamethoxazole,rifampicin,linezolid and fusidic acid,and there was no pvl-positive isolate as well. Conclusions The main pathogen is methicillin-sensitive Staphylococcus aureus（MSSA）in patients with SSTI in Haian area of Jiangsu province. Community-acquired MRSA clone is found in SSTI patients.

Analysis of gut microbiota in obese population with type 2 diabetes mellitus

ZENG Yipeng, HU Yan, WU Ping, FENG Xin'ge, GU Chengying, GUO Yafang

2016, 31(10):  848-853.  DOI: 10.3969/j.issn.1673-8640.2016.010.003

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Objective To investigate the characteristics of gut microbiota in obese population with type 2 diabetes mellitus. Methods A total of 21 obese patients with type 2 diabetes mellitus（study group） and 21 obese subjects with normal glucose（control group） were enrolled. The fecal samples were collected and determined by Illumina Miseq high-throughput sequencing for the V3 region of 16S rRNA gene. The abundance and distribution were analyzed by cluster analysis. Gut microbiota was analyzed by real-time quantitation polymerase chain reaction（PCR）.Results The analysis of DNA sequence was successfully performed. The rare faction curves showed that adequate sequencing depth was achieved,and the coverage indices were >0.99. The Ace and Chao1 indices of study group were lower than those of control group（P<0.05）. The Shannon index or Simpson index of study group was lower or higher than that of control group（P<0.05）. The results of real-time quantitation PCR showed that the quantities of Actinobacteria and Firmicutes were lower（P<0.05）,while that of Bacteroidetes was higher in study group than that in control group（P<0.05）. The quantities of Proteobacteria and Fusobacteria had no statistical significance between the 2 groups（P>0.05）. The quantities ofVeillonella,Clostridium Ⅳ and Bacteroides were higher,and the quantities of Lactobacillus,Blautia coccoides-Eubacterium rectale,Prevotella and Bifidobacterium were lower in study group compared with those in control group（P<0.05）. Conclusions The obese population with type 2 diabetes mellitus has low abundance and diversity of gut microbiota. The quantitation analysis of the composition of gut microbiota has a certain significance.

Distribution of pathogens causing bloodstream infection and in vitro drug susceptibility test result analysis

SHEN Zhenhua, LIU Xinghui, ZHU Hui, YAO Xiaolan

2016, 31(10):  854-857.  DOI: 10.3969/j.issn.1673-8640.2016.010.004

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Objective To understand the distribution characteristic and drug resistance of pathogens in patients with bloodstream infection from Shanghai Pudong Gongli Hospital,and to provide a reference on the choice of antimicrobial agents for treatment. Methods A total of 380 pathogens were isolated from inpatients with bloodstream infection in Shanghai Pudong Gongli Hospital from July 2012 to June 2015,and were identified using VITEK2 Compact automated microorganism analysis system. Drug susceptibility test was performed by VITEK2 Compact. The data were analyzed by WHONET 5.6 software.Results There were 157 isolates of Gram-positive cocci,accounted for 41.3%. Coagulase-negative Staphylococcus,Staphylococcus aureus andEnterococcus were main Gram-positive cocci. A total of 203 isolates of Gram-negative bacilli were isolated and identified,accounted for 53.4%,and Escherichia coli,Klebsiella pneumoniae andPseudomonas aeruginosa were main Gram-negative bacilli. There were 20 isolates of Candida,and Candida albicans was common. The drug resistance rate of Staphylococcus aureus to oxacillin was 47.6%. The drug resistance rates of Pseudomonas aeruginosa and Acinetobacter baumannii to imipenem were 23.1% and 36.4%,and those of Escherichia coli and Klebsiella pneumoniae to ertapenem were 0.0% and 11.1%. The drug resistance rate of Candida albicans to fluconazole was 30.8%. Conclusions Coagulase-negative Staphylococcus and Escherichia coli are the most common pathogens causing bloodstream infection with high drug resistance rates. It should pay more attention to the rational use of antimicrobial agents.

Drug resistance of Helicobacter pylori among children in Shanghai from 2012 to 2014

FU Pan, HE Leiyan, WANG Aimin, SONG Jianming, XUE Jianchang, WANG Chuanqing

2016, 31(10):  858-862.  DOI: 10.3969/j.issn.1673-8640.2016.010.005

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Objective To investigate the drug resistance of Helicobacter pylori （Hp） to 5 antimicrobial agents,which are commonly used in eradication therapy among children in Shanghai.Methods Gastric mucosa biopsy specimens were collected from pediatric patients who had upper gastrointestinal symptoms,and were diagnosed as having Hp infection by rapid urease test. Hp was determined for drug susceptibility by Kirby-Bauer method. All pediatric patients were classified according to sex and age [preschool-age（0-6 years old）and school-age（7-15 years old）],and the drug resistance rates of Hp to metronidazole,clarithromycin,amoxicillin,levofloxacin and moxifloxacin among these groups were compared.Results A total of 359 isolates of Hp were isolated from 743 pediatric patients. The positive rate was 48.3%. The drug resistance rates to metronidazole,clarithromycin,amoxicillin,levofloxacin and moxifloxacin were 49.9%,36.8%,5.3%,5.6% and 7.8%,respectively. No statistical significance in drug resistance rates to the 5 antimicrobial agents was found between boys and girls（P>0.05）. The drug resistance rates of isolates from preschool-age group to metronidazole and amoxicillin（74.1% and 11.1%） were higher than those from school-age group（42.8% and 3.6%,P=0.000 and 0.008）,while isolates against clarithromycin showed an inverse pattern （22.2% in preschool-age group and 41.0% in school-age group,P=0.002）. Conclusions The drug resistance rates of Hp among children in Shanghai are high to metronidazole and clarithromycin. However,the drug resistance rates are low to amoxicillin,levofloxacin and moxifloxacin. Therefore,drug susceptibility should be determined before the 1st time therapy．

Influence of preoperative neutrophil-to-lymphocyte ratio on early recurrence after curative resection in patients with hepatocellular carcinoma

MA Xiaolu, WU Jiong, ZHOU Yan, WANG Beili, ZHANG Chunyan, GUO Wei, PAN Baishen

2016, 31(10):  863-868.  DOI: 10.3969/j.issn.1673-8640.2016.010.006

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Objective To investigate the influence of preoperative neutrophil-to-lymphocyte ratio（NLR） on early recurrence after curative resection in patients with hepatocellular carcinoma（HCC）. Methods The clinical data of 195 HCC patients undergoing curative resection were analyzed retrospectively,and NLR was calculated according to preoperative neutrophil and lymphocyte counts. X-tile software was used to calculate the optimal cut-off value of NLR for tumor recurrence prediction,and the patients were classified into low NLR and high NLR groups. The recurrence prediction values of NLR in 195 patients and low-risk recurrence subgroups were evaluated by Kaplan-Meier curve and log-rank test. Univariate Cox regression model was used to determine the influence of preoperative clinical parameters on predicting recurrence,and multivariate Cox regression analysis was performed to investigate the preoperative clinical parameters with statistical significance.Results For 195 patients,the median follow-up time was 17.30 months,and 43.59% patients（85/195） suffered from tumor recurrence. The optimal cut-off value for NLR was 2.60,and HCC patients were classified into high NLR（≥2.60）group（40 cases）and low NLR（<2.60）group（155 cases）. Univariate analysis showed that preoperative alpha fetoprotein（AFP） >400 ng/mL（P<0.01）,tumor diameter >5 cm（P<0.01）,vascular invasion（P=0.03）,advanced Barcelona Clinic Liver Cancer（BCLC） stage（P=0.01） and high NLR（P<0.01）were influence factors for tumor recurrence free survival rate. Multivariate analysis showed that NLR was the strongest independent factor for tumor recurrence [OR（95%CI）=1.95（1.20-3.19）,P<0.01]. High NLR retained its prognostic value in AFP ≤400 ng/mL,single tumor,no vascular invasion and BCLC stage of 0+A（P<0.05）. High NLR was correlated with low differentiation,big tumor diameter and advanced BCLC stage（P<0.05）. Conclusions Preoperative NLR is a convenient,powerful,simple and low-cost parameter for HCC predicting early recurrence after curative resection. The cut-off value of 2.60 for NLR might be optimal for HCC patients undergoing curative resection in China. The determination of NLR can help with making effective and comprehensive treatment for HCC patients.

Correlations of Lp-PLA2 activity and gene polymorphism with ischemic cerebral stroke

LI Suliang, YE Yun

2016, 31(10):  869-873.  DOI: 10.3969/j.issn.1673-8640.2016.010.007

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Objective To investigate the correlations of lipoprotein-associated phospholipase A2（Lp-PLA2）activity and its gene locus rs1051931（Ala379Val） polymorphism with ischemic cerebral stroke（ICS）. Methods Polymerase chain reaction（PCR） and direct sequencing assay were used to determine rs1051931 polymorphism in 480 patients with ICS and 630 healthy subjects（healthy control group）. Plasma Lp-PLA2 activity,total cholesterol（TC）,triglyceride（TG）,high-density lipoprotein cholesterol（HDL-C）,low-density lipoprotein cholesterol（LDL-C） and glucose（Glu） were determined simultaneously.Results The proportion of males,the proportion of hypertension,Lp-PLA2 activity,TG,TC,HDL-C,LDL-C and Glu in ICS group were higher than those in healthy control group（P<0.05）. The differences of ages and body mass index（BMI） between the 2 groups were not statistically significant（P>0.05）. ICS group had high levels of traditional vascular risk factors,including relatively-high blood pressure and LDL-C and relatively-low HDL-C. After the calibration of traditional vascular risk factors,the highest and lowest quartiles of Lp-PLA2 activity were analyzed {odds ratio（OR）[95% confidence interval（CI）] =2.25（1.23-9.96）}. Lp-PLA2 activities [median（quartile）] were 56.44（27.56-87.84）nmol/（mL·min） in genotype CC,43.26（26.73-78.51）nmol/（mL·min） in genotype CT and 38.24（22.98-62.84）nmol/（mL·min） in genotype TT（P<0.05）. By Binary Logistic regression analysis,Lp-PLA2 genotype was used as an independent variable,the frequency of genotype CC in ICS group was high [OR（95%CI）=1.707（1.055-2.761）]. Conclusions Lp-PLA2 activity in ICS group is higher than that in healthy control group,and its polymorphism is correlated with ICS. Genotype CC is a risk factor for ICS.

Relationship between plasma PCT level and cardiac function in AECOPD patients with heart failure

JI Weixing, ZHOU Xinghui, WANG Xinhua, JIN Huiying, ZHENG Yao, CHEN Xiao

2016, 31(10):  874-877.  DOI: 10.3969/j.issn.1673-8640.2016.010.008

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Objective To investigate the relationship between plasma procalcitonin（PCT） level and cardiac function in acute exacerbation of chronic obstructive pulmonary disease（AECOPD） patients with heart failure（HF）. Methods A total of 40 patients with AECOPD,40 patients with AECOPD combined with HF,40 patients with stable-phase chronic obstructive pulmonary disease（COPD） and 40 healthy subjects were enrolled. AECOPD+HF group was classified into 3 subgroups by New York Heart Association（NYHA） cardiac function classification,including Ⅱ,Ⅲ and Ⅳ subgroups. The levels of plasma PCT and B-type natriuretic peptide（BNP）were determined. The correlation of plasma PCT level and Tei-index was analyzed.Results Plasma BNP and PCT levels from high to low were in AECOPD+HF group,AECOPD group,stable-phase COPD group and healthy control group. There was statistical significance among the groups （P<0.05）. The levels of plasma PCT and BNP in AECOPD+HF group increased with NYHA cardiac function classification,and there was statistical significance among different grades（P<0.05）. Plasma PCT level was positively correlated with BNP and Tei-index（r=0.48 and 0.67,P<0.05）. Multivariate regression analysis showed that PCT was a risk factor for AECOPD {odds ratio（OR）[95% confidence interval（CI）]=1.541（1.361-2.012）}. PCT and BNP were risk factors for AECOPD combined with HF [OR（95%CI）=1.337（1.129-1.534） and 1.656（1.412-2.192）]. Conclusions Plasma PCT level could reflect the severity of cardiac function damage in patients with AECOPD combined with HF.

Change of urinary microalbumin to creatinine ratio in patients with diabetic retinopathy and its significance

JIN Hao, FU Qingping, JU Huixiang, SUN Mingzhong, ZHU Rong, JI Yuqiao, ZHU Hui, JI Jianwei, ZHOU Zhongwei

2016, 31(10):  878-882.  DOI: 10.3969/j.issn.1673-8640.2016.010.009

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Objective To investigate the change of urinary microalbumin（mAlb）to creatinine（Cr）ratio in patients with diabetic retinopathy（DR） and its significance. Methods A total of 173 type 2 diabetes mellitus（T2DM）patients were enrolled,and 40 healthy subjects were enrolled as healthy controls. T2DM patients were classified into 3 groups,non-DR（NDR） group（76 cases）,non-proliferative DR（NPDR） group（58 cases） and proliferative DR（PDR） group（39 cases）. Fasting plasma glucose（FPG）,total cholesterol（TC）,2 h postprandial plasma glucose（2 hPG）,triglyceride（TG）,high-density lipoprotein cholesterol（HDL-C）,low-density lipoprotein cholesterol（LDL-C）,C-reactive protein （CRP）,glycated hemoglobin A_1c（HbA_1c）,urinary mAlb and Cr were determined,and urinary mAlb/Cr ratio was calculated. Simultaneously,related clinical data were collected,including height,weight,waist circumference,hip circumference,the duration of diabetes mellitus and so on. The relationship between urinary mAlb/Cr ratio and other parameters was analyzed by Spearman's rank correlation analysis and multiple linear regression analysis. Logistic regression analysis was used to identify the risk factors of DR.Results Urinary mAlb/Cr ratio increased progressively from NDR,NPDR to PDR groups,and there was statistical significance among the groups（P<0.05）. Urinary mAlb/Cr ratio in T2DM patients was positively correlated with the duration of diabetes mellitus,FPG,2 hPG,LDL-C,HbA_1c and CRP（r=0.372,0.227,0.276,0.231,0.294 and 0.308,P<0.05）. Urinary mAlb/Cr ratio was correlated independently with the duration of diabetes mellitus,CRP and HbA_1c in multiple linear regression analysis（β=0.194,0.169 and 0.183,P=0.007,0.018 and 0.013）. Logistic regression analysis showed that urinary mAlb/Cr ratio,the duration of diabetes mellitus,HbA_1c and CRP were independent risk factors for DR {odds ratio（OR）[95% confidence interval（CI）] 1.212（1.083-1.417）,1.036（1.012-1.063）,1.469（1.140-1.892） and 1.330（1.011-1.273）}. Conclusions Urinary mAlb/Cr ratio is a risk factor for DR,and it is closely related with the development of DR. CRP and HbA_1c may be involved in the development and pathogenesis of DR through damaging renal function.

Relationships of advanced glycation end products and oxidized low-density lipoprotein in elderly diabetic patients with osteoporosis

JIANG Fan, CHEN Xiaozhuang, ZHOU Xiaoli, TONG Shan

2016, 31(10):  883-888.  DOI: 10.3969/j.issn.1673-8640.2016.010.010

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Objective To investigate the relationships of advanced glycation end products（AGE） and oxidized low-density lipoprotein（ox-LDL） in elderly diabetic patients with osteoporosis（OP）. Methods A total of 304 patients with type 2 diabetes mellitus（T2DM）were enrolled,and their ages were ≥60 years. According to the occurrence of OP,they were classified into simple T2DM group（201 cases） and T2DM with OP group（103 cases）. A total of 68 healthy subjects were enrolled as healthy control group. The bone mineral densities of lumbar spine（L1-L4）,spine（except lumbar spine）,left femoral neck（Neck）,left femoral large rotor（Troch） and Ward's triangle area were determined. AGE,ox-LDL,glycated hemoglobin A_1c（HbA_1c）,calcium（Ca）,phosphorus（P）,creatinine（Cr）,urea,fasting plasma glucose（FPG）,2 h postprandial plasma glucose（2 hPG）,uric acid（UA）/ Cr and alkaline phosphatase（ALP） were determined. The data of sex,age,the use of hypoglycemic drugs,the histories of smoking and hypertension,body mass index（BMI） were recorded. Multivariate Logistic regression analysis was used to evaluate the risk factors for T2DM with OP. Spearman correlation analysis was used to analyze the relationship of bone mineral densities with AGE and ox-LDL in T2DM with OP group. Receiver operating characteristic（ROC） curve was used to evaluate the sensitivities and specificities of ox-LDL and AGE for the diagnosis of T2DM with OP.Results HbA_1c,Cr,UA/Cr,FPG,ALP,AGE,ox-LDL and the proportion of hypertension in simple T2DM and T2DM with OP groups were higher than those in healthy control group（P<0.05）. UA/Cr ,AGE and ox-LDL in T2DM with OP group were higher than those in simple T2DM group（P<0.05）,and there was no statistical significance for the other indices（P>0.05）. The bone mineral densities of lumbar spine（L1-L4）,spine,Neck,Troch and Ward's triangle area in T2DM with OP group were lower than those in simple T2DM group（P<0.05）,and the bone mineral densities of lumbar spine（L1-L4）and spine in T2DM with OP group and simple T2DM group were lower than those in healthy control group（P<0.05）. Logistic regression analysis showed that the history of hypertension,UA/Cr,HbA_1c,AGE and ox-LDL were risk factors for T2DM with OP. Spearman correlation analysis showed that AGE and ox-LDL had correlations with the bone mineral densities of Neck,Troch and Wald's triangle area（P<0.05）and no correlation with those of lumbar spine（L1-L4） and spine（P>0.05）. ROC curve showed that the sensitivity,specificity and area under ROC curve of the combined determination of ox-LDL and AGE were higher than those of ox-LDL and AGE single determinations（P<0.05）. Conclusions AGE and ox-LDL have correlations with the incidence of T2DM with OP,and the combined determination of AGE and ox-LDL can improve efficacy for the diagnosis of T2DM with OP.

Determinations of OX40 mRNA,OX40L mRNA and anti-C1q antibody and their diagnosis performance in peripheral blood of patients with lupus nephritis

ZHAO Junfang, LI Guizhen, CHEN Xuewen

2016, 31(10):  889-893.  DOI: 10.3969/j.issn.1673-8640.2016.010.011

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：Objective To investigate OX40 mRNA and OX40L mRNA expressions and anti-C1q antibody level in peripheral blood of patients with lupus nephritis（LN）,and to evaluate diagnosis performance. Methods A total of 40 patients with LN,40 patients without LN and 40 healthy controls were enrolled. OX40 mRNA and OX40L mRNA expressions in peripheral blood mononuclear cells（PBMC）were determined by fluorescence quantitation reverse transcription polymerase chain reaction（PCR）. Anti-C1q antibody level was determined by enzyme-linked immunosorbent assay（ELISA）. Systemic lupus erythematosus disease activity index（SLEDAI）,anti-dsDNA antibody,creatinine and complement were determined.Results OX40 mRNA and OX40L mRNA expressions in PBMC of LN patients were（8±4）×10⁶ copies/μL and（4±3）×10³ copies/μL,which were higher than those in patients without LN [（4±3）×10⁶ copies/μL and（2±2）×10³ copies/μL] and healthy controls [（4±2）×10⁶ copies/μL and（2±1）×10³ copies/μL]（P<0.05）. Anit-C1q antibody level in LN patients [（9.11±2.46） ng/mL] was higher than those in patients without LN[（3.12±0.97）ng/mL] and healthy controls [（1.12±0.33） ng/mL]（P<0.05）. OX40 mRNA expression of LN patients was correlated with anti-C1q antibody,creatinine and SLEDAI（r=0.69,0.75 and 0.67,P<0.01）. There was a correlation between OX40L mRNA expression and anti-C1q antibody level（r=0.58,P<0.01）. Receiver operating characteristic（ROC） curve showed that the sensitivity,specificity and the area under ROC curve（AUC） of OX40L mRNA for the diagnosis of LN were 0.933,0.733 and 0.918,which were better than those of other parameters. Conclusions OX40 mRNA expression and anti-C1q antibody level correlate with the activity of LN,and they may play roles in oncogenesis and progression of LN. OX40 mRNA may become a good parameter for diagnosing and monitoring LN.

The combined determinations of HLA-DR4,anti-CCP antibody and anti-Sa antibody for the diagnosis of rheumatoid arthritis

LI Xiaofeng, LU Xujuan

2016, 31(10):  894-897.  DOI: 10.3969/j.issn.1673-8640.2016.010.012

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Objective To investigate the determinations of human leucocyte antigen（HLA）-DR4,anti-cyclic citrullinated peptide（CCP） antibody and anti-Sa antibody for the diagnosis of rheumatoid arthritis（RA）,and to provide a reference for clinical diagnosis. Methods A total of 148 patients with RA（RA group） and 50 healthy subjects （healthy control group） were enrolled. The levels of HLA-DR4 were determined by sequence specific primer polymerase chain reaction（PCR）,and the levels of anti-CCP antibody and anti-Sa antibody were determined by enzyme-linked immunosorbent assay （ELISA）. Risk factor analysis and the evaluation of combined determinations were performed.Results The positive rates of HLA-DR4, anti-CCP antibody amd anti-Sa antibody in RA group were higher than those in healthy control group, and there were positive correlations of HLA-DR4,anti-CCP antibody and anti-Sa antibody with RA [odds ratios（OR） were 3.28,172.63 and 34.21]. The sensitivities of HLA-DR4,anti-CCP antibody and anti-Sa antibody in RA group were 45.95%,78.38% and 40.54%,and Youden indices were 0.35,0.78 and 0.41,respectively. The Youden indices of the combined determinations of anti-CCP antibody with anti-Sa antibody,HLA-DR4 with anti-CCP antibody and HLA-DR4 with anti-Sa antibody were 0.87,0.74 and 0.54. Conclusions There is a positive correlation of HLA-DR4 with the occurrence and development of RA,and the combined determination of anti-CCP antibody and anti-Sa antibody can improve the diagnosis of RA.

Accuracy of VITEK 2 Compact for the determination of imipenem-resistant Acinetobacter baumannii to amikacin

YANG Yinmei, CHEN Bin, PENG Yingyi, YE Huifen, CHEN Huiling, HUANG Xiaoyuan

2016, 31(10):  907-910.  DOI: 10.3969/j.issn.1673-8640.2016.010.016

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Objective To evaluate the accuracy of VITEK 2 Compact automated microorganism analysis system for the determination of imipenem-resistant Acinetobacter baumannii to amikacin,and to investigate the existence of 16S rRNA methylase gene. Methods The drug susceptibilities of 72 isolates of imipenem-resistant Acinetobacter baumannii and 15 isolates of imipenem-sensitive Acinetobacter baumannii to amikacin were determined by VITEK 2 Compact,Kirby-Bauer method and E-test. 16S rRNA methylase armA gene was determined by polymerase chain reaction（PCR）.Results Among the 72 isolates of imipenem-resistant Acinetobacter baumannii,the drug susceptibility results were consistent between Kirby-Bauer method and E-test. The minimal inhibitory concentration （MIC）of 69 isolates was 4-≥64 μg/mL by VITEK 2 Compact,while the results of Kirby-Bauer method and E-test showed being resistant. The MIC of 3 isolates were ≤2 μg/mL by VITEK 2 Compact,while there were 2 isolates being sensitive by Kirby-Bauer method and E-test. The resistant double-ring phenomenon was found in 59 isolates. The positive rate of carrying armA gene was 92.9% （65/70）by Kirby-Bauer method and E-test. Among the 15 isolates of imipenem-sensitive Acinetobacter baumannii,the MIC to amikacin was ≤2 μg/mL,and the results were in accordance with those of Kirby-Bauer method and E-test. Conclusions VITEK 2 Compact is not reliable for the determination of imipenem-resistantAcinetobacter baumannii to amikacin. The resistant double-ring phenomenon might be related with carrying armA gene.

Application of flow cytometry for the rapid determination of erythrocyte osmotic fragility

WANG Weiwei, YUAN Xiangliang, FEI Qili, SHEN Lisong

2016, 31(10):  911-916.  DOI: 10.3969/j.issn.1673-8640.2016.010.017

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Objective To establish a rapid determination of erythrocyte osmotic fragility by flow cytometry,and to evaluate the feasibility of determining erythrocyte osmotic fragility by flow cytometry. Methods A total of 30 anticoagulation peripheral blood specimens were collected,and each specimen was divided into 2 parts,one group collected within 2 h as normal control group and another group incubated at 37 ℃ for 24 h as positive control group. A total of 3 patients diagnosed as hereditary spherocytosis were enrolled as disease group. The erythrocyte osmotic fragility was determined by flow cytometry. The residue percentage of red blood cells was analyzed by flow cytometry. The optimal volume ratio of normal saline and sterile water was identified. The influence of ethylene diamine tetraacetic acid（EDTA） and heparin anticoagulants was evaluated. A total of 14 cases of heparin anticoagulation specimens were collected,and the red blood cell residue percentage was determined by improved Sanford and flow cytometry simultaneously. The characterization of determining erythrocyte osmotic fragility by flow cytometry and improved Sanford was evaluated.Results By flow cytometry,red blood cell residue percentages were 72.03% ± 5.68% in normal control group and 46.47% ± 15.83% in positive control group（P<0.05）. The disease group showed a significant reduction in red blood cell residue percentage（5.76% ± 3.54%） compared to that in normal control group（P<0.001）. When the volume ratio of normal saline to sterile water was 1.2∶0.8,there was significant difference of red blood cell residue percentage between the 2 groups（P<0.05）. Compared with EDTA anticoagulant,heparin anticoagulant was more suitable for the determination of erythrocyte osmotic fragility. There was a good consistency between flow cytometry and improved Sanford in the determination of erythrocyte osmotic fragility,but flow cytometry had better sensitivity than improved Sanford. Conclusions Flow cytometry has the characteristics of operation simplicity and sensitivity,and has a practical application feasibility in the determination of erythrocyte osmotic fragility.

Research progress on microRNAs in glioma

CHEN Qiudan, LIN Yong

2016, 31(10):  917-921.  DOI: 10.3969/j.issn.1673-8640.2016.010.018

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Glioma is a common malignant tumor in central nervous system with a high lethality. MicroRNAs are known to play vital roles in glioma through regulating the downstream signaling pathways via targets. Therefore,a deep understanding on the roles of microRNAs may provide new insights and theoretical reference for the diagnosis and treatment of glioma in the future.