Table of Content

01 December 2013, Volume 28 Issue 12

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Orginal Article

Changes and correlation study of plasma homocysteine in patients with primary hypertension

YANG Fan,XU Bingxin,GUO Ping,LU Yide.

2013, 28(12):  1069-1072.  DOI: 10.3969/j.issn.1673-8640.2013.12.002

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Objective　 To study the levels of plasma homocysteine(Hcy) in patients with primary hypertension,and to investigate the relationship of Hcy with cardiovascular risk factors and subclinical damage. Methods　 According to the echocardiogram, carotid artery ultrasonic inspection and isotope glomerular filtration rate (iGFR) Chinese Guidelines for the Management of Hypertension, the target damage was analyzed and evaluated (if patients had 2 or more than 2 target damages, they were classified into the groups below and analyzed statistically), and 106 primary hypertension patients were classified into the simple hypertension group (44 patients), the hypertension with left ventricular hypertrophy group (18 patients), the hypertension with carotid atherosclerosis group (46 patients) and the hypertension with decreasing iGFR group (27 patients). Plasma Hcy,fasting plasma glucose (FPG), lipid and creatinine (Cr) were measured, and the results were compared and analyzed by Spearman correlation analysis and multi-logistic regression analysis. Results　 Hcy was positively correlated with subclinical damage and some traditional cardiovascular risk factors [sex, age, smoking, systolic blood pressure (SBP), FPG and Cr](P<0.05).The Hcy levels of the simple hypertension group [(12.18±3.30) μmol/L] were lower than those of the hypertension with decreasing iGFR group[(19.05±8.58) μmol/L], the hypertension with left ventricular hypertrophy group [(18.63±7.99) μmol/L] and the hypertension with carotid atherosclerosis group [(16.80±6.34) μmol/L] (P<0.05). Age,Hcy,FPG and iGFR had statistical significance between the simple hypertension group and the hypertension with carotid atherosclerosis group(P<0.05). Non-carotid atherosclerosis was as dependent variable, and age, FPG, iGFR and Hcy were as independent variables. Multi-logistic regression analysis showed that age was independently associated with carotid atherosclerosis. If only defined Hcy as normal and increasing with the abnormal cut-off value of Hcy >15 μmol/L according to the American Heart Association,the risk of carotid atherosclerosis in hypertension patients with increasing Hcy was 2.74 times higher than than in subjects with normal Hcy. Conclusions　 Age is an important independent risk factor for carotid atherosclerosis in primary hypertension patients. The increasing of Hcy is closely related with subclinical damage,and has a high risk for carotid atherosclerosis.

Analysis on the changes of serum angiotensin-converting enzyme and C reactive protein in liver cirrhosis patients

YAN Feng,XU Weizhen,JIE Weixia,REN Zhenhuan.

2013, 28(12):  1073-1076.  DOI: 10.3969/j.issn.1673-8640.2013.12.003

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Objective　 To investigate the changes of serum angiotensin-converting enzyme (ACE) and C reactive protein (CRP) in liver cirrhosis patients. Methods　 A total of 275 patients with liver cirrhosis and 241 healthy subjects (control group) were enrolled, and their serum ACE activities and CRP levels were determined. The patients with liver cirrhosis were classified into Class A (96 cases), Class B (118 cases) and Class C (61 cases) groups according to liver function Child-Pugh classification standard. ACE activity>65 U/L and CRP level >10 mg/L were as positive judgment values, and the positive rates between the 2 groups were compared. All the subjects were classified into low CRP (≤10 mg/L) group (306 cases) and high CRP (>10 mg/L) group (210 cases), and the prevalence rates between the 2 groups were compared. The 275 patients with liver cirrhosis were classified into high ACE (≥ 65 U/L) group (158 cases) and low ACE (<65 U/L) group (117 cases), and the CRP levels between the 2 groups were compared. Serum ACE activities and CRP levels were analyzed by linear correlation analysis. Results　 The serum ACE activities and CRP levels were higher in Class A, B and C groups than in the control group (P<0.01), and the serum ACE activities and CRP levels increased gradually in Class A, B and C groups(P<0.01). For serum ACE activity>65 U/L as positive judgment value, the positive rate was 74.5% in the liver cirrhosis group, and the positive rate was 5.4% in the control group. For CRP level >10 mg/L as positive judgment value, the positive rate was 68.7% in the liver cirrhosis group, and the positive rate was 8.7% in the control group. The prevalence rate in the high CRP group was 90.0% (189/210), and was 28.1% (86/306) in the low CRP group. The former prevalence rate was 3.2 times than the latter prevalence rate. When the low CRP group was as the reference group, the risk odds ratio (OR) in the high CRP group was 7.937 [95% confidence interval (CI): 6.132-10.530, P<0.01]. The CRP level in the high ACE group [28.6(14.8-86.3) mg/L] was significantly higher than that in the low ACE group [15.5(4.3-42.7) mg/L](P<0.01). Serum ACE activities and CRP levels were positively correlated (r=0.468, P<0.01). Conclusions　 The occurrence and development of liver cirrhosis associate with the changes of ACE and CRP. Inflammation and high ACE status play important roles in occurrence and development of liver cirrhosis.

Evaluation on the application of 6 creatinine-based equations of estimated glomerular filtration rate in healthy population

JIA Keke,YANG Shuo,QIAO Rui,CUI Liyan,ZHANG Jie.

2013, 28(12):  1077-1082.  DOI: 10.3969/j.issn.1673-8640.2013.12.004

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Objective　 To evaluate the application of 6 creatinine-based equations of estimated glomerular filtration rate (eGFR) by enzymatic method and kinetic alkaline picric acid rate method in healthy population. Methods　 Serum creatinine was measured by enzymatic method and kinetic alkaline picric acid rate method in 694 healthy subjects. The eGFR values were calculated and compared by 6 equations [Cockcroft-Gault (C-G) equation, simplified modification of diet in renal disease (MDRD) equation, MDRD-Chinese equation, isotopic dilution mass spectrometry (IDMS)-MDRD equation, chronic kidney disease epidemiology collaboration (CKD-EPI) equation and CKD-EPI for Asian (EPI-Asian) equation]. Results　 Serum creatinine results were(65.8 ± 13.3)μmol/L[(36-117)μmol/L) for enzymatic method,(83.0 ± 12.7) μmol/L[(57-131)μmol/L] for kinetic alkaline picric acid rate method. According to eGFR reference intervals: 85-125mL/min for males and 75-115mL/min for females,if using enzymatic method to detect creatinine,the applicability in healthy subjects of 6 equations were CKD-EPI equation (72.3%)＞IDMS-MDRD equation(69.9%)＞simplified MDRD equation(61.3%)＞EPI-Asian equation (60.7%)＞C-G equation(54.8%)＞MDRD-Chinese people equation (27.3%)(P<0.05),and IDMS-MDRD equation showed the smallest difference between males and females (only 1.6%). If using kinetic alkaline picric acid rate method,the applicabilities in healthy subjects of 4 equations were MDRD-Chinese people equation(80.0%)＞EPI-Asian equation (70.1%)＞CKD-EPI equation (63.8%) ＞ simplified MDRD equation(59.1%)＞C-G equation(52.4%)＞IDMS-MDRD equation(40.7%)(P<0.05),and MDRD-Chinese people equation showed the smallest difference between males and females (only 1%). Conclusions　 There are significant differences among 6 equations. To evaluate eGFR in healthy subjects in North China,CKD-EPI equation and IDMS-MDRD equation are more applicable if using enzymatic mehod traced to IDMS to detect serum creatnine,while MDRD-Chinese people equation is more applicable if using kinetic alkaline picric acid rate method to detect serum creatnine.

The changes of serum CD44v6 and integrin beta 1 before and after cryosurgery in patients with pancreatic cancer and the significance in prognosis evaluation

HUANG Hongli,ZHOU Gang,NIU Lizhi,SHI Hongxia,KONG Xiaofeng,WANG Feng,XU Kecheng.

2013, 28(12):  1083-1087.  DOI: 10.3969/j.issn.1673-8640.2013.12.005

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Objective　 To investigate the changes of the expression levels of serum CD44v6 and integrin beta 1 in patients with pancreatic cancer before and after cryosurgery and the correlation between the expression levels and prognosis. Methods　 Cryosurgery was conducted in 44 patients with pancreatic cancer. Serum samples were collected from the patients before and after cryosurgery and 20 healthy subjects (control group). Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression levels of serum CD44v6 and integrin beta 1. Results　 Compared with the control group,the expression levels of CD44v6 and integrin beta 1 in patients before and after cryosurgery for 10 d were higher (P<0.05). No statistical significance was found between the patients after cryosurgery for 1 month and the control group (P>0.05). The CD44v6 and integrin beta 1 expression levels decreased gradually after cryosurgery. The expression levels of CD44v6 and integrin beta 1 in patients after cryosurgery for 1 month were significantly lower than those in patients before cryosurgery (P<0.05). Both CD44v6 and integrin beta 1 expression levels were significantly correlated with tumor size,tumor differentiation,clinical stage,lymph node metastasis and liver metastasis (P<0.05),while they were not correlated with sex, age and tumor location (P>0.05). The median survival times of patients with low CD44v6 and integrin beta 1 expressions were 14.5 months and 14.2 months,while those of patients with high CD44v6 and intergrin beta 1 expressions were only 9.5 months and 9.8 months. There were statistical significances (P<0.05). Conclusions　 Serum CD44v6 and integrin beta 1 expression levels are closely correlated to the development,invasion,metastasis and prognosis of pancreatic cancer.

Research on the expression of peripheral blood mononuclear cell HLA-G in patients with hepatocellular carcinoma

LIU Yang,HU Mei,YU Hongbo,LI Jian.

2013, 28(12):  1088-1091.  DOI: 10.3969/j.issn.1673-8640.2013.12.006

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Objective 　To investigate the expression level of peripheral blood mononuclear cell (PBMC) human leucocyte antigen G (HLA-G) in patients with hepatocellular carcinoma (HCC). Methods　 The HLA-G mRNA and HLA-G protein expression levels on PBMC from 38 patients with HCC,25 patients with liver cirrhosis and 40 healthy subjects were determined by reverse transcription-polymerase chain reaction (PCR) and flow cytometry (FCM). Results　 The HLA-G mRNA expression rates in HCC group,cirrhosis group and healthy control group were 94.7%,84.0% and 87.5%, respectively,and there was no statistical significance (P=0.359). The HLA-G protein expression rate in HCC group was significantly higher than those in cirrhosis and healthy control groups with statistical significance (P=0.002). In HCC group, the HLA-G protein was mainly expressed on CD4⁺ cells. Conclusions　 The PBMC HLA-G protein expression rate in HCC patients increases,and its mechanism and clinical significance need to be further researched.

The influence of hepatitis B virus on the expression of insulin-like growth factor-binding protein 7 and its clinical significance

SONG Hui,LIU Xinghui,ZHU Chengliang, ZHOANG Jida,LI Yi.

2013, 28(12):  1092-1094.  DOI: 10.3969/j.issn.1673-8640.2013.12.007

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Objective　 To investigate the influence of hepatitis B virus(HBV) on the expression of insulin-like growth factor-binding protein 7(IGFBP7) and its clinical significance. Methods　 The mRNA expression of IGFBP7 was measured by reverse transcription-polymerase chain reaction (PCR), and serum levels of IGFBP7 were measured by enzyme-linked immunosorbent assay(ELISA). The different expressions of IGFBP7 among patients with chronic hepatitis B(CHB), liver cirrhosis(LC) and hepatocellular carcinoma(HCC) were analyzed. Results　 The expression of IGFBP7 mRNA was higher in HepG2.2.15 cells than in HepG2 cells. Compared with healthy controls, serum IGFBP7 levels were much higher in HBV patients (P<0.05), and the serum levels of IGFBP7 were detected at lower levels in patients with LC and HCC than those with CHB(P<0.05). Conclusions　 HBV can upregulate the expression of IGFBP7, which is negatively correlated with the disease progression.

The significance of bone marrow smear microscopy in the diagnosis of AIDS complicated with disseminated Penicillium marneffei

WEI Shanqiu, LUO Xiaolu, LIANG Juanying, FANG Fenghua.

2013, 28(12):  1095-1098.  DOI: 10.3969/j.issn.1673-8640.2013.12.008

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Objective 　To investigate the bone marrow smear microscopy in the diagnosis of acquired immuno-deficiency syndrome (AIDS) complicated with disseminated Penicillium marneffei(PM). Methods　 A total of 257 AIDS patients were enrolled. Bone marrow cell analysis and the peripheral blood and bone marrow fungal dimorphic culture were performed, and the results were analyzed retrospectively. Results　 PM infection was identified in 74 patients by peripheral blood and bone marrow fungal dimorphic culture, and 36 cases were identified as PM infection by bone marrow smear microscopy,among which 3 cases showed negative in fungal dimorphic culture but positive during therapeutic diagnosis. Taking positive fungal dimorphic culture as the standard for PM diagnosis,the specificity of PM by bone marrow smear microscopy was 98.4%, and the sensitivity was 44.6%. The morphology of the bone marrow cell showed that platelet reduced significantly,histiocytosis was reactive, histiocytes were active in phagocytosis,neutrophils developed abnormally or had toxic change,and erythrocytes were morphologically abnormal. The distribution of PM was significantly correlated with the hyperplasia degree of nucleated cell in bone marrow. In the significantly hyperplastic cases,PM were phagocytosed by huge reticulocyte and/or distributed extracellularly. In cases that were hyperplastic,reticulocytes were active in phagocytosis but might not increase,PM could be seen inside or outside of the cells. As for the reduced hyperplastic cases,a small number of PM could be extracellular mainly. The typical characters of PM included sausage shape,elongated cell and a translucent transverse in the middle of the cells before division,and seldom appeared in these cases. Conclusions　 Bone marrow smear microscopy is a highly specific method for the diagnosis of AIDS complicated with disseminated PM. It could facilitate the therapy on time and,sometimes,make up for the omission from the detection of fungal dimorphic culture. It is suitable for the chinical application of AIDS complicated with disseminated PM.

Evaluation of time to positivity for predicting Staphylococci in blood culture

HUANG Lei,SUN Liying,YAN Yan.

2013, 28(12):  1099-1101.  DOI: 10.3969/j.issn.1673-8640.2013.12.009

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Objective　 To investigate the significance of time to positivity (TTP) for predicting Staphylococcus aureus (SA), coagulase negative Staphylococcus (CoNS) and methicillin-resistant Staphylococcus aureus (MRSA) in blood culture. Methods　 The TTP for predicting SA and CoNS in blood culture from 2008 to 2012 in Beijing University First Hospital were collected. MRSA and methicillin-susceptible Staphylococcus aureus (MSSA) were judged by their susceptibility to oxacillin. Only the first isolate was included in this study for those repetitively isolated from the same patients. TTP between SA and CoNS and TTP between MRSA and MSSA were compared statistically. Receiver operating characteristic (ROC) curve was used to analyze the diagnosis significance of TTP. Results　 A total of 101 isolates of SA (51 isolates of MRSA and 50 isolates of MSSA) and 253 isolates of CoNS were collected. TTP of SA and CoNS were (16.63±7.47) h and (24.61±11.09) h with statistical significance (P<0.05), and TTP of MRSA and MSSA were (18.22±8.15) h and (15.01±6.40) h with statistical significance (P<0.05). ROC curve showed that TTP≤16.90 h was the cut-off point to differentiate SA and CoNS, and the area under the ROC curve (AUC) was 0.758. TTP>12.58 h was the cut-off point to differentiate MRSA and MSSA, and the AUC was 0.630. Conclusions　 The accuracy of TTP for predicting SA and CoNS is moderate, and the accuracy of TTP for predicting MRSA and MSSA is low.

Analysis on the resistant characterization, phylogenetic grouping and genetic relatedness of Escherichia coli in urinary tract infection

ZHENG Qisi, CAO Xiaoli, ZHOU Aiping, ZHANG Zhifeng, SHEN Han, ZHANG Kui.

2013, 28(12):  1102-1105.  DOI: 10.3969/j.issn.1673-8640.2013.12.010

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Objective　 To analyze the resistant characterization,phylogenetic grouping and genetic relatedness of Escherichia coli in urinary tract infection, and compare the differences of these factors among the hospital and community infections. Methods　 A total of 185 non-duplicate Escherichia coli in urinary tract infection were analyzed for sensitivities, extended spectrum beta-lactamases (ESBLs) screening, phylogenetic grouping and genetic relatedness, and the results were analyzed statistically. Results　 The resistance rates to multiple antibiotics were>49.0%. A total of 185 isolates displayed a high diversity, and the prevalence of ESBLs was 78.4%. Phylogroup D Escherichia coli was the most frequent(49.7%), and the correlation between the production rate of ESBLs and resistance to cefotaxime was observed (P<0.05). Additionally, there was no statistical significant on the distribution of the factors between the hospital and community infections (P>0.05). Conclusions　 Urinary tract infection is mainly caused by the phylogroup D Escherichia coli which displays a high diversity of genetic relatedness. High-produced ESBLs are multiple resistant to antibiotics.

Performance evaluation of seven cystatin C analysis systems

AN Chongwen,LI Haixia.

2013, 28(12):  1106-1111.  DOI: 10.3969/j.issn.1673-8640.2013.12.011

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Objective　 To evaluate the analysis performance for 7 cystatin C (Cys C ) analysis systems. Methods　 A total of 5 particle-enhanced turbidimetric immunoassay (PETIA) systems and 1 sol particle immunoassay (SPIA) system were evaluated according to the documents of the Clinical and Laboratory Standards Institute (CLSI),and their correlation with particle-enhanced nephelometric immunoassay (PENIA) system from the SIEMENS BNⅡ system (BNⅡ system), precision,accuracy,limit of blank (LoB) and anti-interference ability were evaluated. Results　 For 6 systems (A-F) and BNⅡ system, the total within-run coefficients of variation (CV) were all＜3.75%, and except E, the total between-run CV were all<5%, when Cys C concentration was 0.8-5.0 mg/L. For A-E and BNⅡ systems,accuracy verification showed that the absolute biases for measuring ERM-DA471 were 0.00,0.00,1.01,-0.01,-0.50 and -0.35 mg/L respectively,and the relative biases were 0%,0%,18.43%,-0.18%,-9.12% and -6.39% respectively. The measurement of CAP external quality assessment materials showed that D and BNⅡ systems had desirable results. The LoB were 0.00,0.00,0.31,0.01,0.10,0.06 and<0.05 mg/L for A-F and BNⅡ systems, respectively. When Hb≤13 g/L and TG ≤28 mmol/L,A,B and BNⅡ systems were not significantly influenced (interference＜±10% ). Other systems were influenced with different levels. The A-F systems correlated well with BNⅡ system, and the correlation coefficients (r) were all＞0.975 (P< 0.01). Compared with BN Ⅱ system,the average deviations were -0.02,-0.10,-0.31,0.05,0.04 and 0.36 mg/L for A-F systems, respectively. Conclusions　 The maximum bias can reach 1 mg/L when measuring the reference materials for Cys C analysis systems,and the maximum relative bias is 18.43%. The results of PENIA system are lower than those of PETIA system when measuring ERM-DA471 and CAP external quality assessment materials. In a part of analysis systems, analysis performance is not good.

Evaluation on the reagent performance for the determination of m-AST by enzyme inhibition method and the clinical significance of m-AST in patients with liver disease

LI Jun,XIE Jingsong,ZANG Guizhen.

2013, 28(12):  1112-1115.  DOI: 10.3969/j.issn.1673-8640.2013.12.012

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Objective　 To evaluate the performance of mitochondrial aspartate aminotransferase (m-AST) by enzyme inhibition method, and to investigate the clinical significance of m-AST in patients with liver disease. Methods 　The m-AST activity was determined by enzyme inhibition method, and the activity of cytosolic aspartate aminotransferase (c-AST) was inhibited by protease, and was determined by rate method. The results of enzyme inhibition method were compared with those of immune suppression method. A total of 259 patients with liver disease (43 patients with acute hepatitis, 95 patients with chronic viral hepatitis, 20 patients with liver failure, 11 patients with serious hepatitis, 40 patients with compensated liver cirrhosis, 9 patients with decompensated liver cirrhosis and 41 patients with primary liver carcinoma) and 220 healthy subjects (healthy control group) were enrolled. Serum m-AST activity was measured by enzyme inhibition method and evaluated. The m-AST activities were compared with aspartate aminotransferase (AST) activities. Results　 The within-run coefficient of variation (CV) of enzyme inhibition method was 0.59%-2.23%. The between-run CV was 5.24%-6.23%. The recovery rates were 101.6%-108.0% (the average recovery rate was 104.97%). The linear equation was Y=0.997X-1.51, r=0.999 9. The linearity was up to 450 U/L. The activity of 1500 U/L concentration of c-AST was inhibited completely. The data was positively correlated between enzyme inhibition method and immune suppression method (r=0.999 8). Hemolysis had interference to the m-AST determination results. According to 95% confidence interval (±1.96s), the reference range of m-AST in males was 3.1-9.5 U/L, and that in females was 2.5-8.7 U/L. The activity of m-AST in liver disease group was higher than that in healthy control group (P<0.05), and was positively correlated with AST activity. The ratio of m-AST to AST in healthy control group was 0.30±0.07, and the ratio in liver disease group was lower than that in healthy control group (P<0.05). The decrease degree of m-AST/AST ratio was lower than that of m-AST. Conclusions　 Enzyme inhibition method for m-AST activity had the advantages of high automation, reliability and wide linear range. The m-AST activity can reflect the degree of liver cell injury. It has significance on clinical classification and prognosis of liver disease.

Evaluation on the performance of Destiny Max automatic coagulation analyzer

ZHENG Jiayong, XIA Shuqi, WEI Guozhong, MA Weide, WANG Hai, WANG Han, ZHANG Deting.

2013, 28(12):  1116-1120.  DOI: 10.3969/j.issn.1673-8640.2013.12.013

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Objective　 To evaluate the main performance parameters of Destiny Max automatic coagulation analyzer (Destiny Max analyzer). Methods　 According to the Clinical and Laboratory Standards Institute (CLSI) document, the precision, detection limit, linearity, contamination rate and stability of Destiny Max analyzer were evaluated, and the results were compared with those by Stago STA-R coagulation system (Stago STA-R analyzer). The evaluation parameters included prothrombin time (PT), international standardization ratio (INR), activated partial thromboplastin time (APTT), thrombin time (TT) and fibrinogen (FIB). Results 　The within-run coefficients of variation (CV) of all the results detected by Destiny Max analyzer were <3%,and the between-run CV were <1/3 total error. The detection limit of FIB was 0.50 g/L, and it had good linearity in the range of 0.90-8.97 g/L. The results of contamination rate were less than 1/4 of total error. The results were stable within 15 d after open vial reconstituted. The CV of all the results were <10%. The correlation of Destiny Max analyzer, and Stago STA-R analyzer showed that except the coefficient of correlation (r²) of APTT was 0.813 4,the results of PT,INR and FIB were in high correlation(r²>0.95). Conclusions　 The precision, linearity, detection limit and contamination rate of Destiny Max analyzer meet the requirements. The stability of reagents and performance of Destiny Max analyzer are quite good and suitable for the clinical application.

The application of adenosine triphosphate bioluminescence assay in the diagnosis of multidrug-resistant Mycobacterium tuberculosis

LIU Jun,HU Jiabo, PEI Hao, KUAI Shougang, CHEN Liyan.

2013, 28(12):  1121-1123.  DOI: 10.3969/j.issn.1673-8640.2013.12.014

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Objective　 To compare with Roche solid culture method, and to evaluate the feasibility of adenosine triphosphate bioluminescence assay for detecting multidrug-resistant Mycobacterium tuberculosis. Methods　 By Roche solid culture method and adenosine triphosphate bioluminescence assay, 149 clinical isolates of Mycobacterium tuber-culosis were determined and analyzed. Results　 The coincidence rate of adenosine triphosphate bioluminescence assay with Roche solid culture method was 92.6% (138/149), and the difference had no statistical significance (χ²=0.57, P=0.45). The detection time of adenosine triphosphate bioluminescence assay was (6.6±2.1) d, which was faster than that of Roche solid culture method (28 d, t=422.7, P<0.001). Conclusions　 Compared with the conventional detection methods, adenosine triphosphate bioluminescence assay is simple, rapid and accurate. It is helpful for detecting multidrug-resistant tuberculosis patients and controlling the prevalence of Mycobacterium tuberculosis. It is suitable for clinical laboratory.

Research on the polymorphism of EKLF/KLF1 gene in Lutheran deficient phenotype among Chinese Zhuang population

WANG Chen,JIAO Wei,LI Hailan,YE Luyi, ZHU Ziyan.

2013, 28(12):  1124-1127.  DOI: 10.3969/j.issn.1673-8640.2013.12.015

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Objective　 To reveal the molecule and genetic background of EKLF/KLF1 gene in Lu(a-b-) phenotype among Chinese Zhuang population. Methods　 A total of 22 probands of Lu (b-) phenotype were detected among 4 527 Chinese Zhuang population donors. The Lu (a-b-) individuals of their family members were determined by serologic methods. In addition, 3 exons of EKLF/KLF1 gene were amplified and sequenced. The genetic background was analyzed by sequencing. Results 　Among the 22 probands, 57 cases with Lu (a-b-) phenotype were determined. A total of 51 individuals from 19 pedigrees showed EKLF/KLF1 519-525dupCGGCGCC heterozygous mutation. The other 3 pedigrees′ Lu (a-b-) phenotype had EKLF/KLF1 (895C>G) heterozygous mutation. Conclusions 　The Lu (a-b-) in Chinese Zhuang population might be caused by specific heterozygous mutations in EKLF/KLF1 gene, especially the EKLF/KLF1 519-525dupCGGCGCC heterozygous mutation.

Analysis on the frequency of weak B phenotype and molecular genetics on ABO gene

CHI Quan,ZHANG Ai,REN Benchun.

2013, 28(12):  1128-1131.  DOI: 10.3969/j.issn.1673-8640.2013.12.016

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Objective　 To study the frequency of weak B phenotype and analyze the molecular genetics on ABO gene, and to analyze the serological and genetic characteristics. Methods　 The samples with B phenotypes (moderately agglutinate by anti-B) were identified, screened and collected by the serological techniques in routine ABO blood group. The sequences of exon 6,exon 7and intron 6 of ABO gene were analyzed by direct DNA sequencing. Pedigree study was performed in traceable family. Results 　A total of 13 weak B phenotypes were observed in 241 952 cases (9 cases of blood group B and 4 cases of blood group AB). Enhanced agglutination with anti-H was observed in all the sample′s red cell. The ABO genotypes of A102/B_w12,B101/B101,B101/O02,A102/B101, B101/O01 were detected in 1,2,3,3 and 4 of the 13 cases. A same weak B phenotype was detected in a B101/O01 family (father) study. Conclusions 　The frequency of weak B phenotype in Fujian population with B blood group is about 1∶7 000. A B allele with a nucleotide 278C>T mutation (B_w12) is detected, and no mutation is detected among the others for exon 6, exon 7 and intron 6. Variations outside the sequence of glycosyltransferase catalytic domain may be the factors for weak B phenotype.

Application of six sigma for the performance evaluation of emergency blood parameters at each test phase

WU Xiaohua, SU Lingyun, LI Qiuchen, XIAO Dai, LI Li.

2013, 28(12):  1132-1136.  DOI: 10.3969/j.issn.1673-8640.2013.12.017

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Objective　 To apply six sigma (6σ) for the performance evaluation of emergency blood parameters at each test phase, in order to find out the problems, develop solutions and guide quality improvement. Methods　 The performance evaluation parameters before phase, after phase and during whole process of performance evaluation were determined, and the results from January to December in 2012 were analyzed. A total of 301 125 emergency specimens′ qualification, critical value reporting, report error condition and specimen turn around time (TAT) were analyzed statistically. The sigma (σ) value was calculated. The project performance evaluation was performed in the analysis phase, and the internal quality control data and external quality control data from January to December in 2012 were collected. The imprecision (CV%) and percentage difference were calculated. According to the allowable total error (TEa%), σ value and quality goal index (QGI) were calculated. The parameters unreaching 6σ were analyzed to find out the reasons and guide the quality improvement. Results　 Before, in and after phase and during whole process, the σ values were 5.4, 5.3, 6.1 and 5.4. Conclusions　 6σ can effectively evaluate the emergency blood parameters at each test phase, and manifest the laboratory′s performance level at present and help with finding problems and promoting the continuous improvement of the emergency laboratory quality.

A strain of RSV-A isolated from the monitoring specimens of avian influenza A (H7N9) virus

LOU Yiqun,CHU Wei,SONG Lili,WANG Yu,LAO Baofa.

2013, 28(12):  1137-1139.  DOI: 10.3969/j.issn.1673-8640.2013.12.018

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Objective　 To detect and diagnose respiratory pathogens in the monitoring specimens of avian influenza A (H7N9) virus. Methods　 Nucleic acids of 22 kinds of respiratory pathogens were determined by multiple reverse transcription polymerase chain reaction (RT-PCR). Relative respiratory pathogens were isolated and indentified. Results 　Among the monitoring specimens of avian influenza A (H7N9) virus, nucleic acid of respiratory syncytial virus A (RSV-A) was determined from a severe pneumonia illness patient. The specimen was isolated by MRC-5, and 1 strain of RSV-A was isolated successfully. Conclusions 　Multiple RT-PCR could test 22 kinds of respiratory pathogens simultaneously and contributes to diagnose and control the epidemic situation. The isolation of RSV-A would provide a foothold for the further research of respiratory syncytial virus.