Table of Content

30 August 2013, Volume 28 Issue 8

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Analysis on the results of atypical lymphocyte determined by PENTRA MS60 hematology analyzer

XU Peng,XUE Bingrong,WU Yan, YANG YUwei, XUE Li, YE Ping

2013, 28(8):  659-661.  DOI: 10.3969/j.issn.1673-8640.2013.08.001

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Objective To evaluate the reliability of atypical lymphocyte (ALY)determination results by PENTRA MS60 hematology analyzer (PMS60), and to provide the reference for the review criteria of PMS60. Methods The 193 cases of positive ALY and 215 cases of negative ALY from fresh whole blood samples were detected by PMS60 and prepared for blood smear, and microscopy classification was performed after Wright′s staining. The 408 fresh whole blood samples were analyzed by SYSMEX XS800i hematology analyzer (XS800i) simultaneously. The ALY determination results of the 3 methods were analyzed statistically. Results Compared the results of PMS60 and XS800i, the coincidence rate of positive ALY was 89.7%, and the Kappa value was 0.793. With microscopy classification as the standard, the Kappa value of PMS60 was 0.792 with coincidence, the positive ALY sensitivity was 96.3%, the specificity was 85.3%, the accuracy was 89.7%, and the Yudon index was 0.82. After verification, the accuracy of PMS60 was 94.6%. Conclusions The ALY determination results of PMS60 are in high reliability, and it should perform microscopy classification when ALY proportion>2.5% and / or ALY absolute value>0.25×10⁹/L.

Application on the detection of procalcitonin combined C reactive protein in the early diagnosis of bloodstream infection

TANG Jin,XU Jing,WANG Jianqiang,CHEN Yu,LI Niya,GAO Feng.

2013, 28(8):  662-665.  DOI: 10.3969/j.issn.1673-8640.2013.08.002

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Objective To evaluate the signification of procalcitonin(PCT) combined C reactive protein(CRP) detection in the early diagnosis of bloodstream infection. Methods PCT,CRP,white blood cell(WBC) count and erythrocyte sedimentation rate (ESR) were determined in 156 patients with bloodstream infection,and 105 patients with negative bloodstream infection were as controls.The receiver operating characteristic(ROC)curve was analyzed for PCT,CRP,WBC count and ESR by SPSS 19.0 software.The optimal values were obtained by calculating the area under ROC curve(AUC).The sensitivity and specificity of PCT combined CRP detection in the diagnosis of bloodstream infection were calculated. Results The AUC of PCT,CRP,WBC count and ESR were 0.890,0.714,0.712 and 0.545,respectively.The diagnosis criteria at 0.307 of the ROC curve resulted in the greatest Youden index with the sensitivity of 67.6% and the specificity of 88.6%. The positive and negative predictive values were 82.1% and 78.0%,and the positive and negative likelihood ratio were 5.93 and 0.37. The concordance rate was 79.5%,and Kappa value was 57.4%. When the cut-off value was 0.307 ng/mL, there were statistical significances between the 2 groups(χ²=52.807,P<0.001). The sensitivity of 91.1% and the specificity of 51.7% were in parallel detection of PCT and CRP,and the sensitivity of 55.5% and the specificity of 96.6% were in serial detection. Conclusions PCT is a valuable predictor for the early diagnosis of bloodstream infection.PCT combined CRP detection could improve the sensitivity and specificity.

Evaluation on the performance of VITEK 2C for the susceptibility of Acinetobacter baumannii to amikacin

HUANG Xiaochun,CHEN Yan,LI Yuan, XU Yu, QIN Yanghua

2013, 28(8):  666-670.  DOI: 10.3969/j.issn.1673-8640.2013.08.003

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Objective To evaluate the performance of VITEK^ 2 COMPACT automatic bacterial identification system(VITEK 2C) plus AST GN-13 for the susceptibility of Acinetobacter baumannii to amikacin. Methods Three methods [broth microdilution(BMD), Kirby-Bauer(KB), and VITEK 2C] were used to determine the susceptibility of amikacin to 38 clinical isolates of Acinetobacter baumannii, and then the performances of VITEK 2C and KB were evaluated by using BMD as a reference method. Genes encoding aminoglycoside-modifying enzymes were amplified by multiple polymerase chain reaction(PCR). Results Comparing BMD as the reference method, VITEK 2C plus AST-GN13 yielded major errors in 19 isolates (50.0%) and minor errors in 2 isolates (5.3%). Multiple PCR results of aminoglycoside-modifying enzyme gene were consistent with the results of BMD. Conclusions The VITEK 2C has limitations, which requires that KB should be used for the susceptibility of Acinetobacter baumannii to amikacin.

The etiological survey and epidemiology research of diagrrheagenic Escherichia coli in Xuhui district from 2011 to 2012

ZHAO Xuetao,GAO Kun,ZHANG Chunhua

2013, 28(8):  671-675.  DOI: 10.3969/j.issn.1673-8640.2013.08.004

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Objective To survey the infection situation of diagrrheagenic Escherichia coli(DEC) in intestinal infectious disease, and to analyze the molecular epidemiology characteristics of DEC. Methods The etiological survey was performed continuously through enrolling the cases with gastric and intestinal infectious diseases. The primer probe segements of 10 pairs of virulent genes were determined. The epidemiology distribution characteristics of DEC were analyzed. Results The positive rates of DEC were 9%[including enteroinvasive Escherichia coli(EIEC), enteropathogenic Escherichia coli(EPEC), enterotoxigenic Escherichia coli(ETEC) and enterohemorrhagic Escherichia coli(EHEC)],12%[including enteroaggregative Escherichia coli(EAEC)],17%[including EAEC and diffusively adherent Escherichia coli(DAEC)] and 23%(including EAEC and stable toxin positive). EAEC and DAEC had aging change, and ETEC and DAEC had seasonal fluctuation. DEC was independent of sex . Conclusions The DEC has aging change and seasonal fluctuation. Strengthening the etiological survey of DEC is the foundation of investigating the DEC molecular epidemiology.

Research on serum expression levels of VEGF, TGF-β 1 and endostatin in patients with NSCLC and their correlation

WANG Min

2013, 28(8):  676-679.  DOI: 10.3969/j.issn.1673-8640.2013.08.005

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Objective To determine the serum expression levels of vascular endothelial growth factor (VEGF), transforming growth factor-beta 1(TGF-β1) and endostatin in patients with non-small cell lung cancer(NSCLC), and to investigate the relationship of them with NSCLC clinical and pathological character and the relationship of them to each other. Methods Serum levels of VEGF,TGF-β1 and endostatin were determined by double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) in 63 patients with NSCLC before surgery, and the results were compared with those in 26 patients with benign pulmonary lesion and 28 healthy controls. Results The serum levels of VEGF, TGF-β1 and endostatin in NSCLC group were significantly higher than those in benign pulmonary lesion group and healthy control group ( P<0.01 ). The serum levels of VEGF, TGF-β1 and endostatin were closely correlated with cell differentiation, tumor node metastasis (TNM) stage and metastasis ( P<0.05 ), while they were not correlated with histological type, lymph node metastasis, age, sex and smoking or not( P>0.05 ). The serum levels of TGF-β1 and VEGF were positively correlated in NSCLC group (r=0.456, P<0.01 ), and were negatively correlated between endostatin and VEGF and between endostatin and TGF-β1(r=-0.363 and -0.329,P<0.05). Conclusions VEGF, TGF-β1 and endostatin are closely related with NSCLC angiogenesis and poor cell differentiation. For the formation, development and transfer of NSCLC, VEGF, TGF-β1 and endostatin may be useful indicators for the prediction of lung cancer.

Expressions of SALL4 and Nanog in patients with myelodysplastic syndrome and their clinical significance

DING Liumei,HUANG Hongmei,GAO Song,WU Yangjiong,LI Ying,HUA Fanli

2013, 28(8):  681-684.  DOI: 10.3969/j.issn.1673-8640.2013.08.007

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Objective To investigate the expressions of SALL4 mRNA and Nanog mRNA in peripheral blood and bone marrow in patients with myelodysplastic syndrome(MDS) and their clinical significance. Methods The reverse transcription polymerase chain reaction (PCR) was used to detect the expressions of SALL4 mRNA and Nanog mRNA in the peripheral blood and bone marrow of 75 patients with MDS and the peripheral blood of 30 healthy controls. The difference of the expressions between MDS subtypes was analyzed. Results The expression of SALL4 mRNA in peripheral blood and bone marrow of patients with MDS was significantly higher than that in healthy controls(χ²=22.92,P<0.01). The higher expression of SALL4 mRNA was found in bone marrow than peripheral blood(Z=5.60, P<0.01). The expression of Nanog mRNA was higher in patients with MDS than healthy controls(F=88.78, P<0.01), but there was no difference between peripheral blood and bone marrow(t=0.898, P=0.371). There were statistical significance of SALL4 and Nanog mRNA among MDS subtypes(P<0.01), especially RAEB-Ⅰ and RAEB-Ⅱ. In 7 patients who evolved into acute myeloblastic leukemia,the expressions of SALL4 mRNA(peripheral blood t=2.27, P<0.05; bone marrow t=2.85, P<0.05) and Nanog mRNA(peripheral blood t=5.20,P<0.01; bone marrow t=5.51, P<0.01) were significantly higher than the others. Conclusions SALL4 mRNA and Nanog mRNA are over-expressed in patients with MDS,and the expression correlates with the clinical outcomes.

The analysis of related bone metabolism indicators in postmenopausal osteoporosis

YANG Lishun,YUAN Haisheng,SHEN Xingya,KANG Jianhua,WANG Aiyun.

2013, 28(8):  685-689.  DOI: 10.3969/j.issn.1673-8640.2013.08.008

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Objective To observe the changes of the main factor for regulating osteoclast activity and bone resorption and bone formation markers in vivo of patients with postmenopausal osteoporosis(PMOP), and provide the reference for clinical early prevention of osteoporosis. Methods The levels of serum osteoprotegerin(OPG),nuclear factor-k beta receptor activator ligand(RANKL),bone alkaline phosphatase(BAP),serum tartrate-resistant acid phosphatase(TRACP)-5b,estradiol (E2), plasma homocysteine(Hcy) were determined in 38 patients with PMOP, 50 patients with bone mass reducing and 94 subjects with normal bone mass(control group). Partial correlation analysis was used for some of the indicators excluding age and E2. Independent risk factor analysis and binary Logistic regression analysis were used for all indicators. Receiver operating characteristic (ROC) curve evaluation was used for the diagnosis accuracy. Results A statistical significance in E2,Hcy,OPG,RANKL,TRACP-5b and BAP levels was observed in patient groups and control group(P<0.05, P<0.01, P<0.001). BAP levels of PMOP group were higher than those of bone mass reducing and control groups(P<0.001). However, E2 levels were lower than those of bone mass reducing group(P<0.001) and control group(P<0.05). There was no significant difference in E2 and BAP levels between bone mass reducing group and control group(P>0.05). There was a negative correlation between E2 level with TRACP-5b level(r=-0.198, P=0.007)and BAP level (r=-0.157, P=0.035) excluding age. Excluding age and E2, there was a positive correlation between Hcy with RANKL, TRACP-5b and BAP(r=0.368, P<0.001; r=0.193, P=0.009 and r=0.224, P=0.002) and a negative correlation between Hcy with OPG(r=-0.330, P<0.001). There was a negative correlation between OPG and TRACP-5b(r=-0.458, P=0.008), and there was a positive correlation between RANKL and BAP (r=0.228, P=0.044), excluding age and E2. The areas under the ROC curve of E2, Hcy, OPG, RANKL, TRACP-5b and BAP for the diagnosis of osteoporosis were 0.819, 0.686, 0.803, 0.776, 0.731 and 0.719, respectively. The Logistic regression analysis showed that E2, OPG, TRACP-5b and Hcy had evident effect to osteoporosis excluding age, which the odd ratios(OR) were 0.901, 0.046, 4.165 and 9.112, respectively. Conclusions TRACP-5b, Hcy and BAP are better for the diagnosis of osteoporosis, and they can be used as the reference for the clinical early prevention of PMOP.

The investigation on contents and supplementation situation of trace elements in the whole blood of children

QIN Shan,WANG Yan,YANG Dagui,SONG Kangwen,WANG Kao,ZHANG Qingyi,ZENG Deling.

2013, 28(8):  690-692.  DOI: 10.3969/j.issn.1673-8640.2013.08.009

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Objective To investigate the contents of trace elements and the supplementation situation of trace elements of children in Dujiangyan city, and to guide the parents to add reasonable trace elements to their children. Methods The trace element test results of 3 769 cases of 1 month-old-12 year-old outpatient children were analyzed statistically and reviewed retrospectively,and 200 cases were selected randomly and conducted questionnaire survey. Results The deficiency rates of zinc(30.0%, 1 113 cases), iron(14.7%, 553 cases) and calcium (9.5%, 358 cases) were high. The excessive rates of calcium(25.4%, 964 cases) and zinc(3.9%, 149 cases)were high. The 46% of the 200 cases had been supplemented trace elements by parents[calcium supplementation(85%),zinc supplementation (41%) and iron supplementation (18%)]. Trace element supplementation guided by the media accounted for 51.0%,guided by their own experience accounted for 34.8%, and guided by medical advice accounted for only 18.5%. Conclusions The trace element supplementation of children in Dujiangyan city is not optimistic, and the lack of zinc,iron and calcium is popular. The children do not be supplemented trace elements reasonablely. Children should have regular testing for trace elements, and the diet structure should be adjusted rationally, in order to intensify the children healthcare.

Research on the drug resistance to rifampicin and isoniazid in the detection of Mycobacterium tuberculosis by visible microplate chip

LI Yongjin,OU Qingye,LIU Chunxiao,SHI Lei, ZHAO Chunzhong, GU Libing, XU Yunqing, GU Dayong

2013, 28(8):  693-696.  DOI: 10.3969/j.issn.1673-8640.2013.08.010

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Objective To establish a method based on visible microplate chip for the detection of rifampicin and isoniazid resistance to Mycobacterium tuberculosis. Methods Gene chip was constructed by immobilizing default mode probes against genes associated with rifampicin and isoniazid resistance to Mycobacterium tuberculosis on the surface of gene chip. Drug resistance was identified by the prepared gene chip to detect the relative gene mutation, and the positive results,a color dot on the surface of gene chip,can be observed by naked eye. Results The prepared visible microplate chip showed stable detection results. The 40 of 50 (80%) strains were resistant to isoniazid with a high mutation site of 315 codon within katG gene, and 15 of 20 (75%) strains were resistant to rifampicin with a high mutation site of 531 and 526 codons within rpoB gene. The results from the gene chip detection were consistent with the polymerase chain reaction(PCR)sequencing analysis. Conclusions The prepared visible microplate chip with good specificity and sensitivity is a fast, accurate and convenient mothod, and is promising for the detection of rifampicin and isoniazid resistance to Mycobacterium tuberculosis.

Methodology comparison of serum creatinine determined by isotope dilution liquid chromatography tandem mass spectrometry,enzymatic method and Jaffe method

SONG Yunxiao,OU Meixian,LI Shuijun,ZHANG Haichen,YU Chen.

2013, 28(8):  698-703.  DOI: 10.3969/j.issn.1673-8640.2013.08.012

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Objective To compare the results of the serum creatinine determined by isotope dilution liquid chromatography tandem mass spectrometry (ID-LC-MS/MS), sarcosine oxidase enzymatic method (enzymatic method) and Jaffe method, and provide the reference for concordance from different determination systems. Methods Serum creatinine levels in 200 clinical serum samples, 50 hemolyzed samples, 50 lipidic samples (triglyceride 1.88-17.60 mmol/L) and 6 standard serum samples were determined by ID-LC-MS/MS, enzymatic method and Jaffe method. The precisions, recoveries, relative biases and interferences to hemolysis and lipid were compared among the 3 methods. Results Adding 50 μmol/L and 100 μmol/L creatinine, the coefficients of variation (CV) were < 1.14%, < 2.39% and < 3.84%, the recoveries were 84.9%, 82.2%; 74.4%, 70.8% and 96.1%, 96.3% for enzymatic method, Jaffe method and ID-LC-MS/MS, respectively. Compared to ID-LC-MS/MS, the linearity regression equations of enzymatic method and Jaffe method were Y_{enzymatic method}=0.964X_ID-LC-MS/MS+0.385(r=0.994) and Y_{Jaffe method}=0.955X_ID-LC-MS/MS+13.14(r=0.979). Compared to ID-LC-MS/MS, the average relative bias of serum creatinine were -2.93% by enzymatic method and 13.9% by Jaffe method. Significant negative interferences were observed in hemolyzed and lipidic samples. Conclusions There is a good comparability on serum creatinine between enzymatic method and ID-LC-MS/MS. The serum creatinine level can be overestimated by Jaffe method. The serum creatinine levels can be underestimated in hemolyzed and lipidic samples by enzymatic method and Jaffe method.

Study on the new sensor chip-differential potentiometric stripping analysis in the detection of blood lead

TANG Yin,SHENG Qingsong,ZHANG Tongjun,XIN Junwei.

2013, 28(8):  704-706.  DOI: 10.3969/j.issn.1673-8640.2013.08.013

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Objective To evaluate the accuracy and repeatability of clinical blood lead detection by sensor chip-differential potentiometric stripping analysis and compare with hydride generation atomic fluorescence spectrometry, and to provide the reference for clinical application. Methods The 2 methods were used to analyze the blood samples and the certified reference materials (national blood lead detection external quality assessment samples in 2011). The accuracy, repeatability, recovery rate,acceptable range and coincidence rate of sensor chip-differential potentiometric stripping analysis were evaluated. Results All of the results of 6 certified reference materials were in the allowable error range(target value ?10%), and the accuracy was good. The relative standard deviations (RSD) of the low,medium and high blood lead level blood samples were 6.98%,4.09% and 2.78%, respectively.The average recovery rate was 98.1%, and the system ratio error was 1.9%. Compared the results of sensor chip-differential potentiometric stripping analysis and hydride generation atomic fluorescence spectrometry, the results had no statistical significance(P>0.05), and the correlation coefficient of linear regression analysis(r²) was 0.964. The coincidence rate of the 2 methods was 96.4%. Conclusions sensor chip-differential potentiometric stripping analysis has good accuracy, repeatability and precision, and is suitable for the clinical application of detecting blood lead.

Assessment of new iodide ion-selective electrode for the determination of urine iodine

WANG Jun, ZHENG Jingyu.

2013, 28(8):  707-710.  DOI: 10.3969/j.issn.1673-8640.2013.08.014

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Objective To assess the application significance of the new iodide ion-selective electrode for the determination of urine iodine. Methods Performance parameters and selectivity coefficients of the new iodide ion-selective electrode were measured according to the national mechanical industry standard JB/T9362-1999 Technical Specifications for the Ion-Selective Electrode. National standard reference materials(GBW09108-GBW09110) were performed for the new iodide ion-selective electrode to evaluate the accuracy. Urine iodine ion concentrations of 126 urine samples were determined by the new method, and the results were compared with the national health care industry standard method-arsenic cerium catalytic spectrophotometric determination(WS/T107-2006). Results The detection range of the new iodide ion-selective electrode was 50-690 μg/L with a slope ≥ 55 mV, the time to response < 2 min, the applicable pH range 2-10, the internal resistance < 500 kΩ, repeatability error < 2 mV, stability <±8 mV/24 h and the range of applicable temperature 5-60 ℃. The selectivity coefficients of the new iodide ion-selective electrode to Cl^-, S^2- and etc. reached 10^-7 grade. The interference of co-existing organic materials was eliminated. The determination results of the national standard reference materials were within the standard value range.There was no statistical significance between the results of urine iodine ion concentrations of 126 urine samples by the new iodide ion-selective electrode and standard methods(t=0.527,P=0.604). There was a good correlation between the 2 methods, and the linear regression equation was Y=0.972 9X＋4.484 5(r=0.978 7). Conclusions The new iodide ion-selective electrode can be used for the determination of urine iodine ion.

The study of rate method to detect the activity of diamine oxidase with double-reagent

HU Jinfang,WU Yan,WANG Baishi,WANG Xu.

2013, 28(8):  711-715.  DOI: 10.3969/j.issn.1673-8640.2013.08.015

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Objective To establish a continuous monitoring method to detect the activity of diamine oxidase(DAO) with double-reagent, and to evaluate methodologically. Methods The method was used to identify the optimum reaction conditions about the specific absorption peak,the optimum detection time,a reasonable ratio of reagents,the optimum substrate concentration, the best pH value of reaction buffer system and the concentration of the buffer. The sensitivity,accuracy and anti-interference capability were evaluated by linear range,precision,recovery, interference test and stability of reagents and samples. The reference interval and diagnostic accuracy were identified by detecting the activity of DAO in 150 healthy subjects and 30 patients who were severely burnt and combined with gastrointestinal mucosa barrier dysfunction or failure. Results The final reaction buffer system was based on the 100 mmol/L Tris(hydroxy-methyl)-aminomethane hydrochloride (Tris-HCl) buffer (pH 8.0). The optimum substrate was 3.2 mmol/L 1,4-tetranethylenediamine, and the ratio of reagent Ⅰ and reagent Ⅱ was 4∶1 . The activity of DAO was calculated by the continuous monitoring of the lowering speed of nicotinamide adenine dinucleotide(NADH) absorbance under the wavelength of 340 nm during 200-300 s. The linear range was 0-100 U/L, the imprecision was < 5%, the recovery rate was 95.8% and 98.0%,the method was not interfered when bilirubin＜150 μmol/L,hemoglobin＜4 g/L,lipid＜4 g/L,NH₄⁺＜60 μmol/L and monoamine oxidase(MAO)＜120 U/L. The reagents were stable in 2 weeks. The normal activity of serum DAO was (3.90 ± 2.96)U/L,and according to 95% confidence interval, the diagnostic reference range was is 0 - 9.7 U/L. The area under receiver operating characteristic(ROC) curve was 0.960, and the diagnostic accuracy was good. Conclusions The continuous monitoring method to detect the activity of DAO with double-reagent is sensitive and reproducible,and has strong ability of anti-interference and good diagnostic accuracy. The stability of reagent can meet the daily needs of automatic biochemical analyzer for detection. It may become a routine clinical detection method for the diagnosis of intestinal mucosa injury.

Determination and its significance of DNMT mRNA expression in Hep3B,HepG2 and hepatocellular cancer tissues

GAO Bo,LI Haiping,YU Zongtao,Li Jun,ZHANG Jicai.

2013, 28(8):  716-719.  DOI: 10.3969/j.issn.1673-8640.2013.08.016

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Objective To investigate the expression of DNA methyltransferase(DNMT) mRNA in Hep3B, HepG2 and hepatocellular cancer tissues,and to analyze the relationship between hepatitis B virus(HBV) infection and DNMT mRNA expression. Methods Fluorescence quantitative polymerase chain reaction (PCR) was used to determine the mRNA expressions of DNMT1, DNMT2, DNMT3A, DNMT3B mRNA and GAPDH genes in Hep3B, HepG2 and hepatocellular cancer tissues. Results In HBV-associated cancerous tissues,the mRNA expression levels of DNMT1,DNMT 3A and DNMT 3B mRNA were higher than those in non-HBV-associated cancerous tissues (P<0.05). The expressions of DNMT1, DNMT2, DNMT3A and DNMT3B mRNA were 1.26?0.15, 0.76?0.15, 1.26?0.20 and 0.66?0.13 in Hep3B and 0.64?0.11, 0.62?0.12, 0.48?0.12 and 0.36?0.10 in HepG2. The expressions of DNMT1, DNMT3A and DNMT3B mRNA in Hep3B were higher than those in HepG2(P<0.01). Conclusions Overexpression of DNMT mRNA is detected in HBV-associated Hep3B, HepG2 and hepatocellular cancer tissues. The HBV infection can stimulate the expression of DNMT mRNA,and involve the methylation of tumor-suppressor genes.

Continuous evaluation on the unpassed items in proficiency testing and improvement on the determination quality of outpatient department and emergency department

WANG Jie,Li Yuan

2013, 28(8):  720-722.  DOI: 10.3969/j.issn.1673-8640.2013.08.017

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Objective To evaluate continuously the unpassed items in proficiency testing(PT), improve the determination quality of outpatient and emergency departments, and provide clinical accurate report. Methods The College of American Pathologists(CAP) PT reports were evaluated, and the unpassed items were analyzed and improved into the quality management plan. Results The determination numbers of items in PT and unpassed rates from 2007 to 2011 were 2 440, 2.6%; 2 593, 1.1%; 2 355, 0.6%; 1 894, 0.5%; and 1 799, 0.4%. The differences of unpassed rates from 2007 to 2011 had statistical significance (P< 0.05). The main factors for unpass were methodological, technical and clerical errors. Conclusions During the last 5 years, the PT unpassed rates decrease significantly. CAP accreditation is beneficial for the continuous quality improvement.