Table of Content

01 November 2014, Volume 29 Issue 11

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Orignal Article

Challenge on the clinical examination for gravidas and children

FU Qihua, YANG Haiou.

2014, 29(11):  1081-1083.  DOI: 10.3969/j.issn.1673-8640.2014.11.001

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With the rapid development in medicine and related fields, advanced determination technologies and novel clinical indicators are emerging rapidly. As for the distinct characteristics of gravidas and children, the demands for clinical indicators and determination technologies are different from those in adults. There will be an important challenge on the clinical examination for gravidas and children, such as normal reference interval establishment and the standardization and clinical rational explanation for novel clinical indicators and advanced determination technologies.

The diagnostic significance of neutrophil gelatinase-associated lipocalin in acute renal injure after cardiopulmonary bypass in pediatric cardiac surgery

XU Jiayin, XU Jin.

2014, 29(11):  1084-1088.  DOI: 10.3969/j.issn.1673-8640.2014.11.002

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The review summarizes the diagnostic characteristic of neutrophil gelatinase-associated lipocalin and its diagnostic significance in acute renal injure after cardiopulmonary bypass in pediatric cardiac surgery compared to other diagnostic markers.

Application of multiplex polymerase chain reaction for detecting the serotypes of Streptococcus pneumonia

PAN Fen, ZHANG Hong

2014, 29(11):  1089-1093.  DOI: 10.3969/j.issn.1673-8640.2014.11.003

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Streptococcus pneumoniae is one of common pathogens causing pediatric infections. Owing to the characteristic of capsular polysaccharide, Streptococcus pneumoniae can be differentiated into 40 serogroups and more than 90 serotypes by conventional capsular quellung reaction. Recently, with the development of molecular biological techniques, multiplex polymerase chain reaction (MP-PCR) gradually replaces the conventional capsular quellung reaction, which is used to detect the serotypes of Streptococcus pneumoniae. Furthermore, MP-PCR has the advantages of easy and rapid operation and low costing, which could provide the basis for monitoring the prevalent serotypes of Streptococcus pneumoniae in clinic.

An investigation on the reference range of glycosylated hemoglobin A_1c in neonatal umbilical cord blood

ZHANG Li, YU Jing, WANG Jieyun, GU Qin, LI Haichuan, HOU Yaping, GONG Bo

2014, 29(11):  1094-1096.  DOI: 10.3969/j.issn.1673-8640.2014.11.004

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Objective　To investigate the reference range of neonatal glycosylated hemoglobin A_1c (HbA_1c). Methods　A total of 1 003 healthy infants were enrolled, and their blood samples from umbilical cord were collected. HbA_1c levels were determined by affinity chromatography, and were calculated according to P_2.5-P_97.5. Results　The HbA_1c levels showed a positive skewness distribution. The median of HbA_1c levels was 3.80%. The 95% reference ranges for HbA_1c levels were 3.46%- 4.25%. There was no significant influence of infants' sex and delivery ways on HbA_1c levels (P>0.05). There was a positive correlation between HbA_1c levels and the blood glucose of infants (r=0.118, P=0.028) and no correlation with the weight of infants (r=0.064, P=0.168). Conclusions　HbA_1c levels can help for monitoring the blood glucose of infants, and the reference range has an important significance in clinic.

Preliminary investigation on the establishment of biological reference intervals about the main parameters of blood cells among pregnant women

GE Yafang, SHEN Yunyue, TANG Zhenhua

2014, 29(11):  1097-1100.  DOI: 10.3969/j.issn.1673-8640.2014.11.005

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Objective　To establish the biological reference intervals about the main parameters of blood cells among pregnant women in our laboratory. Methods　A total of 331 pregnant women who had normal and term delivery were enrolled. According to their medical records, their 2 reports of pregnancy routine blood cell tests were obtained. The results were analyzed statistically, and the biological reference intervals were calculated by SAS 8.1 software. Results　There was statistical significance in all parameters of pregnancy routine blood cell tests between the first and third trimesters (P<0.05). The results of biological reference intervals showed that the 2 sets of data about platelet (PLT), monocyte percentage (M %) and basophilic granulocyte percentage (B %) were all within current biological reference intervals, then the current biological reference intervals could be used for pregnancy. The 2 sets of data about hematocrit (HCT) and eosinophile granulocyte percentage (E %) could be combined to new biological reference intervals. The biological reference intervals for red blood cell count (RBC), hemoglobin (Hb), white blood cell count (WBC), neutrophile granulocyte percentage (N %) and lymphocyte percentage (L %) should be established according to different trimesters. Conclusions　The newly established biological reference intervals are appropriate to reflect the physiological changes during pregnancy, and can guide the clinical diagnosis and treatment effectively.

Influence of maternal weight on MoM and screening performance in second trimester prenatal triple marker screening

ZHANG Bin, LIU Xiaoyan.

2014, 29(11):  1101-1106.  DOI: 10.3969/j.issn.1673-8640.2014.11.006

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Objective　To verify the validity of maternal weight adjustment, and to study the influence of maternal weight adjustment on second trimester prenatal triple marker screening in the Obstetrics and Gynecology Hospital of Fudan University. Methods　A total of 28 577 second trimester pregnant women were enrolled, and their serum samples were determined by chemiluminescence immunoassay for Down syndrome second trimester prenatal screening with triple markers, alpha fetoprotein (AFP), beta subunit of human chorionic gonadotropin (β-hCG) and unconjugated estriol (uE3). After regressing the maternal weight adjustment parameters, the MoM of maternal weight adjustment was obtained, and the risks of Down syndrome, Edward syndrome and open neural tube disease were calculated. Results　Reciprocal-linear was the best fitting model. The reciprocal-linear model detected 2 cases of Down syndrome, thus the detection rate increased from 65% to 76%, comparing to that of no maternal weight adjustment. Conclusions　 Maternal weight adjustment can increase the detection rate of Down syndrome as well as diminish the influence of maternal weight on serum marker concentrations. Every laboratory should establish a laboratory-specific maternal weight adjustment to report accurate risk assessment for Down syndrome, Edward syndrome and open neural tube disease.

Analysis of variants with unknown significance indicating 9p23 as a candidate locus for intellectual development impairment

LIU Hongjing, YAO Ru'en, YU Yongguo, SUN Fenyong

2014, 29(11):  1107-1111.  DOI: 10.3969/j.issn.1673-8640.2014.11.007

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Objective　To find out copy number variants as candidate loci by analyzing variants with unknown significance (VUS) and phenotypes in chromosomal microarray analysis (CMA). Methods　The VUS and phenotypes in chromosomal microarray for 38 patients were analyzed statistically and classified into different groups (intellectual level, development level, facial abnormality and head circumference abnormality) to evaluate the relation between VUS and phenotypes. A total of 45 healthy controls were used in the study to validate polymorphism copy number variants. Results　Deletions on 9p23 region had significantly higher frequency in intellectual development impairment patients than patients with normal intellectual level with statistical significance (P<0.01), and 9p23 region change had not been detected in 45 healthy controls, indicating this locus to be related with children's intellectual development impairment. Conclusions　The analysis of VUS can offer the reference between phenotypes and copy number variants. The confirmation of VUS can help with enhancing the diagnostic ability of chromosomal microarray and find new pathogenic chromosomal locus.

The correlation analysis between fibrinogen and preeclampsia

SUN Li, YU Qiongyan, CHEN Ying, LI Dashuai, YU Jing, GONG Bo

2014, 29(11):  1112-1114.  DOI: 10.3969/j.issn.1673-8640.2014.11.008

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Objective　To investigate the pathogenetic role and clinical significance of fibrinogen in preeclampsia. Methods　The medical records of 85 healthy pregnant women, 47 mild preeclampsia pregnant patients, 27 severe preeclampsia pregnant patients and 55 healthy women without pregnancy reviewed retrospectively. The predictive value of fibrinogen levels was assessed by receiver operating characteristic (ROC) curve. Pearson method was used to analyze the correlation between fibrinogen and the time of preeclampsia onset. Results　Preeclampsia pregnant patients had higher plasma fibrinogen, compared with either healthy pregnant women or healthy women without pregnancy (P<0.01). The areas under ROC curve of fibrinogen for mild and severe preeclampsia [95% confidence interval (CI)]were 0.70 (0.62-0.78) and 0.70 (0.58-0.82). Negative correlation was observed between plasma fibrinogen and the time of preeclampsia onset (R＝－0.33，P<0.01). Conclusions　Plasma fibrinogen is helpful for preeclampsia prediction.

Establishment and evaluation of fluorescence isothermal amplification assay for the detection of Enterovirus 71

CAO Lingfeng, SU Liyun, DONG Niuniu, XU Jin.

2014, 29(11):  1115-1119.  DOI: 10.3969/j.issn.1673-8640.2014.11.009

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Objective　To establish a rapid and reliable fluorescence isothermal amplification assay (SAT) for the detection of Enterovirus 71 (EV71). Methods　By designing specific primers and optimal probe and using M-MLV reverse transcriptase and T7 RNA polymerase, the SAT was developed for detecting EV71 based on isothermal RNA amplification and fluorescence quantitation polymerase chain reaction (PCR). A total of 199 stool specimens from the children with hand, foot, mouth disease from Children's Hospital of Fudan University were determined. Using EV71 PCR fluorescence diagnostic kits as reference kit and DNA sequencing as the third party verification method, the Kappa analysis was performed on the study data for consistency. Results　A total of 119 stool specimens were positive by SAT, and there were 21 specimens being negative by PCR-fluorescence probe assay, but 20 specimens were confirmed to be positive by DNA sequencing. The Kappa value between SAT and PCR-fluorescence probe assay was 0.789. The sensitivity and specificity of SAT were 100% and 98.77%, respectively. Conclusions　This study indicates that RNA SAT for the detection of EV71 is a sensitive, specific, accurate, rapid and reliable method, and it should be a potential method for the rapid detection of EV71 infection.

The analysis of IgE and eosinophil determinations in acute and chronic urticaria children

CAI Defeng, LU Yuanshan, YUAN Yan, YANG Xiaolin, WU Yueping, MA Dongli

2014, 29(11):  1120-1123.  DOI: 10.3969/j.issn.1673-8640.2014.11.010

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Objective　To investigate IgE levels, eosinophil change and their clinical significance in acute and chronic urticaria children. Methods　Respectively by chemiluminescence, enzyme-linked immunofluorescence and volume, conductivity and scattering (VCS) principle, serum total IgE levels, food, mites and plant-specific IgE levels and eosinophil percentages were determined, and the samples were collected from 77 acute and 46 chronic urticaria children. The differences between acute and chronic urticaria children were analyzed comparatively. Results　There was no statistical significance in total IgE levels and positive rates to food, mites and plant-specific IgE levels between acute and chronic urticaria children with other allergic diseases(P>0.05), but there was a significantly higher total IgE level in chronic group than that in acute group in acute and chronic urticaria children without other allergic diseases (P<0.05). Boys had significantly higher serum total IgE levels than girls. The total IgE levels had a trend of increasing with age. There was a higher positive rate to mites-specific IgE than food and plant-specific IgE(P<0.05). The eosinophil percentages of acute and chronic urticaria children had no statistical significance (P>0.05). Conclusions　Eosinophil percentage is not a good index for differential diagnosis in acute and chronic urticaria children. IgE has a certain significance for diagnosing acute and chronic urticaria without other allergic diseases. However, in other cases, serum level of total IgE is not a good index for differential diagnosis in acute and chronic urticaria children. Integrating case history, course of disease and clinical symptom can help with making a correct diagnosis.

Clinical significance of serum pepsinogen and anti-Hp IgG antibody in the diagnosis of peptic ulcer

ZHAO Lanjing, LIU Chunxing, AN Xianyuan.

2014, 29(11):  1124-1127.  DOI: 10.3969/j.issn.1673-8640.2014.11.011

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Objective　 To investigate the clinical significance of serum pepsinogen (PG) and anti-Helicobacter pylori (Hp) IgG antibody in the diagnosis of peptic ulcer. Methods　A total of 115 peptic ulcer patients (peptic ulcer group) were found by attending physical examination from October 2007 to December 2012 in Huadong Sanatorium of Shanghai, including 65 cases of gastric ulcer and 50 cases of duodenal ulcer. A total of 90 healthy subjects were enrolled as controls, and the sex and age ratios were the same with those in peptic ulcer group. The levels of serum PGⅠ and PGⅡ were measured by latex-enhanced immunoturbidimetry, and the PGⅠ/Ⅱratio(PGR)was calculated. The serum anti-Hp IgG antibody was determined by enzyme-linked immunosorbent assay (ELISA). Results　In peptic ulcer group, the serum levels of PGⅠand PGⅡand PGR were (180.14±20.56) ng/mL, (24.98±10.14) ng/mL and 7.87±1.29, respectively, which were significantly higher than those in control group (P<0.05).The positive rate of anti-Hp IgG antibody infection in peptic ulcer group was 77.4%, and that in control group was 23.3%, with statistical significance(P<0.05).Serum PGⅠ and PG Ⅱlevels and PGR in HP positive group were (165.35±22.14) ng/mL, (20.75±11.26) ng/mL and 7.64±2.03, respectively, which were significantly higher than those in HP negative group (P<0.05). Conclusions　The serum PG and anti-Hp IgG antibody are related closely with peptic ulcer changes, and they can be used for the early screening of peptic ulcer.

The detection result analysis on the immune functions in cirrhosis patients with liver cancer and patients with liver cancer alone

ZHANG Yichao, XIA Jun, LI Xiong

2014, 29(11):  1128-1131.  DOI: 10.3969/j.issn.1673-8640.2014.11.012

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Objective　To investigate the changes of cellular and humoral immune functions in cirrhosis patients with liver cancer and patients with liver cancer alone. Methods　A total of 40 cirrhosis patients with liver cancer and 40 patients with liver cancer alone were enrolled. At the same period， 40 healthy subjects with no liver disease history were enrolled as controls. Their peripheral blood samples were extracted， and their cellular immune level changes were determined by flow cytometry， including T lymphocyte subset（CD3， CD4， CD8 and CD4 / CD8） values. The concentrations of humoral immunity（IgA， IgG and IgM） were determined by immunoturbidimetry. Results　The CD3， CD4 and CD4/CD8 significantly decreased in liver cancer patients than in controls（P＜0.05）， and the CD3， CD4 and CD4/CD8 in cirrhosis patients with liver cancer decreased obviously compared with those in patients with liver cancer alone（P＜0.05）. Compared with those in controls， the IgA， IgG and IgM increased obviously in liver cancer patients， and the IgA， lgG and lgM increased obviously in cirrhosis patients with liver cancer compared with those in patients with liver cancer alone（P＜0.05）. Conclusions　The immune functions of cirrhosis patients with liver cancer decrease more significantly than those of patients with liver cancer alone. Therefore， improving the immune functions in patients with liver cancer and rebuilding the defense system are so important， and monitoring the changes of immune functions has a positive role for estimating prognosis and guiding clinical treatment.

Monitoring serum bactericidal activity determination in patients with renal insufficiency undergoing antibacterials

DONG Chenglin, XU Ling, SHAO Yan, CUI Bangquan, LIU Zhihui, HONG Huifeng

2014, 29(11):  1132-1135.  DOI: 10.3969/j.issn.1673-8640.2014.11.013

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Objective　To monitor serum bactericidal activity（SBA） determination in renal insufficiency patients during antibiotics treatment， to adjust the antibiotics dosage and dosing interval， to formulate individualized dosing regimen， and to improve the treatment and decrease the side effects. Methods　A total of 48 renal insufficiency [glomerular filtration rate（GFR）＜50 mL/min] patients undergoing cephalosporin treatment were enrolled， including 36 cases with middle renal insufficiency and 12 cases with severe renal insufficiency， their antibiotics dosage and dosing interval were adjusted according to the guideline of antibiotics treatment for renal insufficiency patients， and the peak and valley SBA values were measured by the method we designed. Results　In 36 patients with middle renal insufficiency， the peak SBA value was 1∶32-1∶128（median 1∶64）， and the valley SBA value was 1∶8-1∶64（median 1∶32）. In 12 patients with severe renal insufficiency， the peak SBA value was 1∶64-1∶512（median 1∶128）， and the valley SBA value was 1∶16-1∶128（median 1∶64）. The SBA values in the 2 groups were all higher than reasonable suggestion value（P＜0.01）. Conclusions　In patients with renal insufficiency undergoing antibiotics treatment mainly metabolized by kidney， there will increase the side effects and renal insufficiency. It should monitor SBA value， when using antibiotics mainly metabolized by kidney in patients with renal insufficiency.

Drug suspectibility and homology analysis of carbapenem-resistant Klebsiella spp

SUN Kangde, CHEN Xu, YU Zhongmin, CHEN Fuxiang

2014, 29(11):  1136-1140.  DOI: 10.3969/j.issn.1673-8640.2014.11.014

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Objective　To investigate the resistance mechanism and epidemiology of carbapenem-resistant Klebsiella spp， and to establish the pulsed-field gel electrophoresis（PFGE） typing database of carbapenem-resistant Klebsiella spp， in order to provide the reference for rapidly source-tracking and controlling the bacterium spread at the outbreak of nosocomial infection. Methods　Carbapenem-resistant Klebsiella spp.were collected，and the phenotypic and genetic features were detected by modified Hodge test and polymerase chain reaction（PCR）， respectively. PFGE typing was performed for homology analysis. Results　The main resistance mechanism of carbapenem-resistant Klebsiella spp. was Klebsiella pneumoniae carbapenemase（KPC）-producing， and there were 2 genotypes by PFGE typing， including genotype A（4 isolates） and genotype B（4 isolates）. Conclusions　KPC-producing is the main reason for carbapenem-resistant Klebsiella spp. in the hospital， and it is necessary to monitor the resistance and to strengthen the infection control.

Clinical distribution and drug resistance analysis for 63 isolates of Stenotrophomonas maltophilia

XU Fei, LIU Xuemei, CHI Fuli, TAN Hua, CHANG Yong, CAO Bo

2014, 29(11):  1141-1143.  DOI: 10.3969/j.issn.1673-8640.2014.11.015

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Objective　To investigate the clinical distribution and drug resistance of Stenotrophomonas maltophilia. Methods　The clinical distribution and drug resistance analysis for 63 isolates of Stenotrophomonas maltophilia（K-B） method. The results were analyzed statistically. Results　The isolation rate was the highest in respiratory specimens . There was a highest isolation rate of 87.3%. The samples were mainly obtained from the wards of Neonatology Department（17.4%） and Thoracic Surgery（17.4%）， followed by Respiratory Department（4.3%） and Intensive Care Unit（ICU）（14.3%）. The 63 isolates of Stenotrophomonas maltophilia， and the resistance rate was 100.0%. The resistance rates to levofloxacin， minocyline and triethoprim-sulfamethoxazole were 1.6%， 3.2% and 3.2%. Conclusions　It should enhance drug resistance monitoring of Stenotrophomonas maltophilia. Treatment based on drug susceptibility test should be adapted as soon as possible. were highly resistant to imipenem were performed by VITEK-2 COMPACT system and Kirby-Bauer.

Liquid chromatography-tandem mass spectrometry for the monitoring of theophylline concentration in serum and its application to external quality assessment

SUN Xuying, ZHANG Meiwei, LI Shuijun

2014, 29(11):  1144-1150.  DOI: 10.3969/j.issn.1673-8640.2014.11.016

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Objective　To perform the methodology evaluation of a liquid chromatography-tandem mass spectrometry（LC-MS/MS） for the determination of theophylline in serum， and to investigate the application significance in the therapeutic drug monitoring（TDM） for theophylline. Methods　After being added with radionuclide theophylline-D6 as internal standard， serum samples were treated with protein precipitation and determined by LC-MS/MS. A reverse phase chromatographic separation was performed on Capcell C18 MG Ⅲ analytical column（100 mm×2.0 mm， 5 μm） by using 0.1% formic acid in acetonitrile and 0.1% formic acid in water [20∶80（v/v）] as mobile phase. The flow rate was 0.3 mL/min. Theophylline and internal standard were monitored by a positive electrospray ion-tandem mass spectrometry system. The proficiency performance of LC-MS/MS was tested by participation to external quality assessment of theophylline TDM for the National Center for Clinical Laboratory. Results　The linear range of LC-MS/MS for the determination of theophylline was 1-50 μg/mL. The accuracy was 94.14%-104.00% with within-run and between-run precisions of 2.26%-6.65% and 4.70%-6.84%. The run time was 3.5 min per sample. Stabilities were good under 3 cycles of frozen-thaw for 3 times from -30 ℃ to room temperature， room temperature for 24 h， autosampler for 24 h and -30℃ for 28 d. The external quality assessment proficiency test showed the bias of 2.75%， regression slope of 1.04 and correlation coefficient（r²） of 0.983. Conclusions　The radionuclide internal standard-dilution LC-MS/MS is simple， rapid， specific and sensitive for the determination of serum theophylline. It is comparable with the external quality assessment proficiency test for consecutive 6 years， and it is suitable for theophylline TDM in clinical practice.

Significance of HbA_1c and GA in initial screening of gestational diabetes mellitus

ZHANG Yexin, WANG Haijun, HU Hao, ZHU Xiangyu.

2014, 29(11):  1151-1153.  DOI: 10.3969/j.issn.1673-8640.2014.11.017

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Objective 　To investigate the significance of glycated albumin（GA） and glycosylated hemoglobin A_1c（HbA_1c） in screening gestational diabetes mellitus（GDM）. Methods　There were 289 women with pregnancy for 22-28 weeks. A total of 202 cases were enrolled in normal blood glucose group， and 87 cases were enrolled in GDM group. Hexokinase method was used to detect glucose， and high pressure liquid chromatography was used to detect HbA_1c，and enzyme method was used to detect GA. Results　HbA_1c of GDM group was 5.20%±0.09%， which was significantly higher than that of normal blood glucose group（5.03%±0.02%，P＜0.01）. GA of GDM group was 13.48%±0.28%， that of normal blood glucose group was 13.39%±0.09%， and the difference had no statistical significance（P＞0.05）. Pearson correlation analysis showed that glucose was positively correlated with HbA_1c（r=0.203，P＜0.01）， and there was no correlation between glucose and GA（r=0.114，P＞0.05）. There was no correlation between HbA_1c and GA（r=0.041，P＞0.05）. Using HbA_1c 95% value（5.50%） in normal blood glucose group as the judgment basis for screening GDM， the specificity was 94.55%， and the sensitivity was 86.21%. Conclusions　HbA_1c， other than GA， is a good indicator for screening GDM initially.

Correlation analysis of serum osteopontin and liver fibrosis in non-alcoholic fatty liver disease patients

QIAN Niandong, HUANG Ping, QI Jin, DENG Lianfu.

2014, 29(11):  1154-1157.  DOI: 10.3969/j.issn.1673-8640.2014.11.018

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Objective　To investigate the significance of serum osteopontin（OPN） level in the diagnosis of liver fibrosis in non-alcoholic fatty liver disease（NAFLD）. Methods　A total of 86 NAFLD patients and 60 healthy controls（healthy control group） were enrolled. Liver biopsy was carried out in NAFLD patients to evaluate liver fibrosis stages. Enzyme-linked immunosorbent assay（ELISA） was used to detect serum levels of OPN in NAFLD patients and healthy controls， and the correlations of OPN level with aspartate aminotransferase（AST）， alanine aminotransferase（ALT）， total cholesterol（TC）， high-density lipoprotein cholesterol（HDL-C）， low-density lipoprotein cholesterol（LDL-C）， triglyceride（TG） and high-sensitivity C reaction protein（hs-CRP） were analyzed. Results　Compared with healthy controls， body mass index（BMI）， systolic pressure， diastolic pressure， AST， ALT， homeostasis model assessment for insulin resistance（HOMA-IR）， TC， LDL-C and TG in NAFLD patients significantly increased， and the proportion of diabetes mellitus and metabolism syndrome was higher（P＜0.01）. Serum OPN level was significantly higher in NAFLD patients and showed positive correlation with AST， ALT， hs-CRP and liver fibrosis stages（r=0.224， 0.265， 0.192 and 0.219， P＜0.001， ＜0.001， =0.003 and ＜0.001， respectively）. There were negative correlations with sex and HDL-C（r=-0.213 and -0.191， P＜0.01 and =0.041， respectively）. In addition， for liver fibrosis stage ≥3 in NAFLD patients， when the cut-off value was 32 μg/L， the area under receiver operating characteristic（ROC） curve was 0.897， and the sensitivity and specificity were 0.821 and 0.855. Conclusions　As a pro-fibrotic factor， OPN can not only predict NAFLD fibrosis stages， but also plays a role in pathogenesis of NAFLD.

The significance of comprehensive analysis in the diagnosis of leukemia

HUANG Jingqin, XU Shanshan, LI Zhi, ZHENG Te, WENG Wenhao, WANG Jiayi

2014, 29(11):  1158-1163.  DOI: 10.3969/j.issn.1673-8640.2014.11.019

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Objective　To testify the significance of comprehensive analysis in the diagnosis of leukemia. Methods　A total of 6 cases of leukemia， including acute promyelocytic leukemia M3a， large granular lymphocyte leukemia， hairy cell leukemia（HCL）， acute myeloblastic leukemia with significant maturation M2a， chronic lymphocyte leukemia（CLL） and acute myeloblastic leukemia without cytologic maturation M0 were diagnosed by routine blood testing， bone marrow morphology， cellular chemical dyeing， flow cytometry， cellular genetics， molecular biology analysis and bone marrow pathological diagnosis. Results　Acute promyelocytic leukemia was confirmed through bone marrow morphology and cellular chemical dyeing. Large granular lymphocyte leukemia was confirmed through bone marrow morphology，cellular chemical dyeing and flow cytometry. HCL was confirmed through bone marrow morphology， cellular chemical dyeing， flow cytometry and bone marrow pathological diagnosis. However， the diagnosis of acute myeloblastic leukemia with significant maturation manifested that it was still hard to make the decision through bone marrow morphology， cellular chemical dyeing and flow cytometry. The confirmation was made in aid of chromosome examination and molecular biology analysis. CLL was confirmed through bone marrow morphology and flow cytometry. Acute myeloblastic leukemia without cytologic maturation was confirmed through bone marrow morphology， cellular chemical dyeing， flow cytometry and chromosome examination. Conclusions　The comprehensive analysis of multiple laboratory items makes critical clinical importance in the confirmation of untypical leukemia as the results of environmental changes，aging of population and so on.

The changes of Zn, Fe and blood routine test in children with infection and anemia

CAI Mei, TANG Zhenhua, YU Wen.

2014, 29(11):  1164-1168.  DOI: 10.3969/j.issn.1673-8640.2014.11.020

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Objective　To investigate the relationship of trace element zinc（Zn）， iron（Fe） and blood routine test with infection and anemia in children in order to determine and perform trace element supplementary reasonably. Methods　A total of 1 289 children aged 1 to 5 years were enrolled. The 287 children with infection were selected as infection group， and 303 children with anemia were selected as anemia group. The 699 healthy children were selected as control group. The peripheral blood levels of trace elements and blood routine test were determined. Results　The level of peripheral blood Zn in infection group [（63.8±5.6）μmol/L] was lower than that in control group [（69.6±6.3）μmol/L] with statistical significance（P＜0.05）. The level of peripheral blood Fe in infection group [（7.74±0.26）mmol/L] had no statistical significance with that in control group（P＞0.05）. The levels of white blood cell（WBC） [（10.9±2.4）×10⁹/L] and 5% neutrophilic granulocyte（NEUT） [（37.9±14.5）%] in infection group were higher than those in control group . The level of peripheral blood Fe in anemia group [（7.68±0.23）mmol/L] was lower than that in control group [（7.80±0.26）mmol/L] with statistical significance（P＜0.01）， and the level of peripheral blood Zn in anemia group [（65.1±6.3）μmol/L] had no statistical significance with that in control group（P＞0.05）. Hemoglobin（Hb） in anemia group[（110.5±6.8）g/L] was lower than that in control group [（127.4±5.6）g/L， P＜0.05]. The contents of red blood cell（RBC）， erythrocyte mean corpuscular volume（MCV）， mean corpuscular hemoglobin（MCH） and mean corpusular hemoglobin concentration（MCHC） in anemia group were lower than those in control group with statistical significance（P＜0.01）. The level of Fe in anemia group was lower than that in infection group with statistical significance（P＜0.05）. Conclusions　The changes of trace elements in peripheral blood of children with infection and anemia are significant， especially Zn and Fe， with the realtionship of blood routine test.

Optimization and application of performance evaluation protocol for syphilis serologic assays

GU Weiming, YANG Yang, WU Lei, GUO Wei, SONG Binbin, DENG Anmei, WANG Qingzhong

2014, 29(11):  1169-1174.  DOI: 10.3969/j.issn.1673-8640.2014.11.021

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Objective　To design and optimize performance evaluation protocol for syphilis serologic assays， and to investigate the significance of new protocol for discovering the characteristics of assay performance. Methods　Technical parameters， such as lower limit of detection， interference and stability were introduced firstly into the optimized performance evaluation protocol， and 6 domestic and imported assay reagents for syphilis serology were evaluated according to the protocol by multi-center joint study. Results　The qualitative test for 456 definite specimens showed 100% agreement result between point-of-care testing and clinical diagnostic test， and exhibited various degrees of false positive and false negative results among the other 4 screening assays， although their sensitivity and specificity had no statistical difference. The lower limits of detection of the reagents descended from high to low in the order of screening assays were treponema pallidum particle agglutination assay， point-of-care testing and enzyme-linked immunosorbent assay/chemiluminescence immunoassay. Two brands of procoagulant vacuum blood collection tubes interfered with non-specific antibody detection reagent for syphilis in a 96.88% proportion. Conclusions　The study shows that the performance of domestic syphilis serologic assays is comparable with that of imported ones. The result by point-of-care testing exhibits a high agreement with clinical diagnostic test result， and its sensitivity is better than that of the traditional method， which lays a foundation for spreading in clinical application. The optimized performance evaluation protocol for syphilis serologic assays would reflect assay performance better.

Application and evaluation of Vitek 2 Compact system to identify clinical rare bacteria

HUANG Lin, GU Chi, TANG Huqiang

2014, 29(11):  1175-1177.  DOI: 10.3969/j.issn.1673-8640.2014.11.022

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Objective　To retrospectively analyze the identification process of strain 1304 in external quality assessment, and to evaluate the application of Vitek 2 Compact automatic microorganism analyzer system（Vitek 2 Compact system） to identify clinical rare bacteria. Methods　Strain 1304 was identified comprehensively by traditional microscopic morphology, manual biochemical reaction and Vitek 2 Compact system, simultaneously. The final result was confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry（MALDI-TOF MS）. Results　Strain 1304 was Gram-negativebacilli. The result of oxidase test was positive.The first result wasBordetella bronchiseptica（identification probability was 99%）. The second result was low discrimination（51%Alcaligenes faecalis and 49%Pseudomonas pseudoalcaligenes） by Vitek 2 Compact system. The result of supplementary test（NO₃） was negative. The final identification result wasAlcaligenes faecalis, and MALDI-TOF MS got the same result with a score value of 2.158.Conclusions　It exists some limitations in the identification of rare bacteria by Vitek 2 Compact system. Comprehend means should be used to decrease wrong identification rate.

Research on the mechanism of less susceptibility to cefepime than to ceftazidime in clinical isolates of Pseudomonas aeruginosa

ZENG Zhangrui, WANG Weiping, HUANG Mei, SHAO Haifeng

2014, 29(11):  1178-1183.  DOI: 10.3969/j.issn.1673-8640.2014.11.023

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Objective　To investigate the reasons of less susceptibility mechanism to cefepime (FEP) than to ceftazidime (CAZ) in clinical isolates of Pseudomonas aeruginosa. Methods　A total of 60 isolates ofPseudomonas aeruginosa which had been tested being less susceptible to FEP than to CAZ for minimal inhibitory concentration (MIC) by Vitek 2 Compact automatic microorganism analyzer system were collected. Agar dilution method was used to reexamine the MIC of FEP and CAZ. The resistance genes were amplified by polymerase chain reaction (PCR). The expressions of efflux system were analyzed by real-time fluorescence quantitation PCR. Results　The 5% of error rate was showed between agar dilution method and Vitek 2 Compact system. There were isolates with KPC and PSE-1 resistance genes by PCR amplification. There were mainly the expressions of mexB and mexD in efflux system. The sequencing of regulatory genes showed that there were 3 isolates for mexB which the Gly70(GGG) ofnalC was substitute for Glu(GAG)(GGG→GAG) and 2 isolates for mexD which the Gly109(GGC) of nfxB was substitute for Val (GTC)(GGC→GTC). Conclusions　Less susceptibility to FEP than to CAZ in clinical isolates ofPseudomonas aeruginosa is due to the production of KPC and PSE-1 resistance genes and the increasing expression levels of mexAB-OprM and mexCD-OprJ.

Research on the inhibition on Klebsiella pneumoniae clinical isolates by antimicrobial peptide human β defensin 3

ZHANG Lei, YIN Shuwei, ZHANG Ting, QI Xin, LÜ Huiying, LIN Ping

2014, 29(11):  1184-1187.  DOI: 10.3969/j.issn.1673-8640.2014.11.024

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Objective　To analyze the inhibition effects of antimicrobial peptide human β defensin 3（hBD3） against Klebsiella pneumoniae clinical isolates. Methods　Antimicrobial peptide hBD3 was synthesized. The direct inhibition on 20 Klebsiella pneumoniae clinical isolates were detected by minimal inhibitory concentration（MIC） test， antibacterial activity test and the analysis on some important functional genes. The antimicrobial peptide was combined with amoxicillin， ceftazidime and ciprofloxacin for observing the effects on antibiotic 50% MIC（MIC50） and 90% MIC（MIC90）. Results　The MIC of hBD3 against Klebsiella pneumoniae clinical isolates was（22.3±6.6）μg/mL， the bactericidal activity was observed while hBD3 concentration was as high as 8 μg/mL. The hBD3 could improve the expression of ompC and reduce the expression of yojL.When 5 μg/mL hBD3 was added ， the MIC50 and MIC90 against Klebsiella pneumoniae clinical isolates of ceftazidime and ciprofloxacin were dropped down. Conclusions 　There is a considerable antibacterial activity of antimicrobial peptide hBD3 against Klebsiella pneumoniae clinical isolates. It could be used to inhibit Klebsiella pneumoniae infection combined with antibiotics.

Research on lysyl oxidase and pelvic organ prolapse

WU Huiheng, CHEN Xinliang, TONG Xiaowen

2014, 29(11):  1191-1194.  DOI: 10.3969/j.issn.1673-8640.2014.11.026

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Pelvic organ prolapse (POP) is a common gynecopathy, but the pathogenesis of POP is obscure up to now. Recent studies have shown an association between POP and the destroy of structural and functional integrity of pelvic connective tissue. The main proteins composing pelvic connective tissue extracellular matrix (ECM) are elastin and collagen. Elastin and collagen contents and structural changes can cause the decreasing of tissue elasticity and the relaxation of pelvic floor structure. Lysyl oxidase (LOX) can catalyze the polymerization of elastic fibers forming covalent crosslinking to increase the stability of EMC, so it is supposed that the change of LOX expression may make a certain contribution to the occurrence and development of POP.

Experience exchange of quality improvement in common coagulation function determinations

WANG Beili, PAN Baishen

2014, 29(11):  1195-1198.  DOI: 10.3969/j.issn.1673-8640.2014.11.027

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Coagulation function determinations are usually used to analyze the causes of bleeding disorders, evaluate pre operation coagulation status and support the therapies of oral coagulation drugs. China National Accreditation Service for Conformity Assessment, College of American Pathologists and Clinical and Laboratory Standards Institute published the terms and documents of quality assurance in coagulation function determinations. Considering the account of coagulation specimens, clinical laboratories may ignore some quality requirements, affect clinical diagnosis and treatment, and influence on patients' safety of life. The review mainly focuses on the common problems and their solutions of clinical coagulation function determinations.