Table of Content

30 March 2013, Volume 28 Issue 3

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The determination and its clinical significance of serum GM-CSF，IL-6，IL-17 and TNF-α in patients with rheumatoid arthritis

TANG Ronghua，HUANG Jianjun

2013, 28(3):  173-177.  DOI: 10.3969/j.issn.1673-8640.2013.03.001

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Objective　To determine the concentrations of serum granulocyte-macrophage colony stimulating factor(GM-CSF)，interleukin 6(IL-6)，interleukin 17(IL-17) and tumor necrosis factor-alpha(TNF-α)，and investigate their significance in the pathogenesis of rheumatoid arthritis(RA). Methods　A total of 81 RA outpatients were enrolled，including 36 inactive RA patients and 45 active RA patients，and 50 healthy subjects were as controls. Disease activity score 28(DAS28) of RA patients was evaluated，and the concentrations of serum GM-CSF，IL-6，IL-17 and TNF-α were determined by enzyme-linked immunosorbent assay(ELISA). The results were expressed as median(inter-quartile range). Results　The concentrations of serum GM-CSF were 12.7(9.7) pg/mL in inactive RA group，27.4(34.8) pg/mL in active RA group and 6.2(5.2) pg/mL in control group. The concentrations of serum IL-6 were 8.5(10.5)，47.1(43.8) and 4.7(5.3) pg/mL in the 3 groups，respectively. The concentrations of serum IL-17 were 19.3(16.4)，23.4(19.5) and 20.8(16.1) pg/mL in the 3 groups，respectively. The concentrations of serum TNF-α were 42.7(37.2)，76.2(42.4) and 34.1(21.9) pg/mL in the 3 groups，respectively. The concentrations of serum GM-CSF，IL-6 and TNF-α were higher in active RA group than in inactive RA group and control group(P＜0.01). The concentrations of serum GM-CSF，IL-6 and TNF-α were higher in inactive RA group than in control group(P＜0.05). IL-17 showed no statistical significance among the 3 groups(P＞0.05). GM-CSF was significantly correlated with IL-6 and TNF-α (r＝0.637 and 0.738 ，P＜0.001). There was no correlation between GM-CSF and IL-17 (r＝0.171，P＝0.426). In RA patients，there were correlations of DAS28 with GM-CSF，IL-6 and TNF-α (r＝0.583，0.656 and 0.758，P＜0.001). Conclusions　The concentrations of serum GM-CSF，IL-6 and TNF-α play important roles in RA pathogenesis. GM-CSF is a potential role in the regulation of IL-6 and TNF-α in RA patients.

The changes and significance of clinical routine laboratory test results in hepatitis B patients before and after undergoing liver transplantation

JI Qiang，GU Xing，ZHOU Jun，CHEN Dongmei，YIN Yuepeng，GAO Chunfang

2013, 28(3):  178-182.  DOI: 10.3969/j.issn.1673-8640.2013.03.002

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Objective　To analyze the changes of clinical routine laboratory test results in patients with hepatitis B before and after undergoing liver transplantation，and investigate the regularity and characteristics before and after undergoing liver transplantation and in the period of recovery.  Methods　A retrospective study was performed in 44 patients with preoperative antiviral therapy undergoing liver transplantation. Alanine aminotransferase (ALT)，aspartate aminotransferase (AST)，total bilirubin (TBil)，direct bilirubin (DBil)，total bile acid (TBA)，alkaline phosphatase (ALP)，gamma-glutamyltransferase (GGT)，total protein (TP)，albumin (Alb)，prealbumin (PAlb)，glucose (GLU)，prothrombin time (PT)，activated partial thromboplastin time (APTT)，platelet (PLT) and hepatitis B virus (HBV) DNA load and other clinical routine laboratory tests were determined before and 1，10，20，30 d after undergoing liver transplantation. The results were expressed as median(range).  Results　Before the operation，AST and ALT were 70.2(15.5-539.6) IU/L and 44.1(5.8-255.5) IU/L，and they increased sharply to 1 506.4(172.4-5 195.3) IU/L and 749.1(142.2-2 874.2)IU/L after 1d，and then quickly dropped to 117.3(17.2-900.4) IU/L and 135.3(27.5-1 237.1) IU/L after 10 d，and ended up with the normal levels after 20 d. GGT and ALP decreased slightly after 1 d，and they increased after 10 d，and then remained high after 30 d. TBil and DBil mostly continued to decline，and came back to normal after 30 d. TP and Alb slightly decreased after 1 d，and returned to be normal after 10 d. PAlb continued to increase after the operation. TBA decreased after the operation. PT and APTT were prolonged after the operation，and returned to be normal after 10 d. PLT declined after the operation，and returned to be normal after 10 d. Except for 2 patients who had no change in serum hepatitis B markers and HBV DNA loads between preoperation and postoperation，the remaining 42 patients became negative in serum hepatitis B surface antigen (HBsAb) and turned positive in serum anti-hepatitis B surface antigen antibody (anti-HBs antibody)，and their HBV DNA loads became lower than the limit of detection. Conclusions　Liver transplantation combined with comprehensive antiviral and immunological treatments can induce the seroconversion of HBsAg and hepatitis B e antigen(HBeAg)，and has significant reduction of HBV DNA detection rate and hepatitis B recurrence rate. The variations and characteristics of perioperative routine laboratory test results in patients undergoing liver transplantation may perform as a basis for clinical condition judgments.

Establishment of the reference range of fibronectin for children aged 0-14 years old in Shaoxing region

GE Guoxing，ZHONG Yaping，LIANG Meichun

2013, 28(3):  183-185.  DOI: 10.3969/j.issn.1673-8640.2013.03.003

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[Abstract] Objective To study the value of fibronectin (Fn ) among healthy children (aged 0-14 years) in native,and to establish the children's medical reference range. Methods The 353 children aged 0-14 years whose liver, kidney, heart and lung functions were normal were selected as subjects,in which there were 229 males and 124 females. The sample was divided into three groups: 0-1 years,1-3 years and 4-14 years. Fn was detected in the ABBOTT Aeroset full-automatic biochemical Analyzer ( American company Abbott ) with immune turbidimetry principle and with biochemical reagents producted by Shanghai North biochemical reagents company,and the detection results were processed according to CLSIC28-A2 (clinical laboratory reference ranges are defined and confirmed-2nd Edition). Results The fibronectin reference range in healthy children was 122.8-250.2 μg/ml, 125.5-270.7 μg/ml, 128.9-279.4 μg/ml in male and 125.2-289.5μg/ml,130.3-280.0μg/ml, 132.6-262.1μg/ml in female. Conclusions The laboratory is necessary to establish its own reference range according to the CLSIC28-A2 documents,in order to provide reference to different clinical departments.

Analysis on the results of cervical disease liquid-based cytology and human papillomavirus detection in 708 migrant female workers in Fengxian District，Shanghai

GU Qilong 1，FENG Minhua 2

2013, 28(3):  186-188.  DOI: 10.3969/j.issn.1673-8640.2013.03.004

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Objective　To investigate the prevalence of cervical diseases and high-risk human papillomavirus (HR-HPV) infection in migrant female workers in Fengxian District，Shanghai.  Methods　The results of liquid-based cytology and HR-HPV detection of 708 migrant female workers seeking medical advice for possible cervical diseases in Fengxian District，Shanghai were analyzed retrospectively. The results of biopsy and loop electrosurgical excisional procedure (LEEP) from a part of these patients were also analyzed.  Results　Among these migrant female workers，the prevalences of atypical squamous cell (ASC)，low-grade squamous intraepithelial lesion (LSIL) and high-grade squamous intraepithelial lesion (HSIL) were 31.6%，23.2% and 11.9%，respectively. The prevalence of HR-HPV was 68.4%. With the increase of age，the prevalences of LSIL and HR-HPV decreased，but the prevalence of HSIL increased. Cervical intraepithelial neoplasia or worse (CIN2+) results were found by biopsy in 35.7% of HR-HPV positive ASC patients and 37.8% of HR-HPV positive LSIL patients，respectively. Among HSIL patients，85.7% were CIN2+ by biopsy or LEEP.  Conclusions　The prevalence of cervical diseases is high among migrant female workers in Fengxian District，Shanghai. Some of them have high-grade lesions.

Detection of erm genes and epidemiology research of Staphylococcus aureus in patients with bloodstream infection

WANG Lixin 1，HU Yiming 2，HU Zhidong 1，TIAN Bin 1，LI Jing 1，WANG Fengxia 1，YANG Hua 1

2013, 28(3):  189-193.  DOI: 10.3969/j.issn.1673-8640.2013.03.005

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Objective　To investigate the distribution and epidemic situation of erythromycin ribosome methylase（erm）genes in erythromycin-resistant Staphylococcus aureus in patients with bloodstream infection from January 2006 to August 2011. Methods　A total of 39 isolates of erythromycin-resistant Staphylococcus aureus were collected from January 2006 to August 2011 among patients with bloodstream infection. The bacterium identification and antimicrobial susceptibility test were conducted by VITEK 2 Compact automatic system，matched identification card and antimicrobial susceptibility card. Methicillin-resistant Staphylococcus aureus (MRSA) was identified by disk diffusion method with cefoxitin. Phenotypic expression of inducible resistance was assessed by D test. Polymerase chain reaction (PCR) was used to detect ermA，ermB and ermC genes. The epidemic situation of ermA-producing Staphylococcus aureus was analyzed by pulsed-field gel electrophoresis (PFGE) and Staphylococcal cassette chromosome mec(SCCmec)typing. Results　The resistant rates of 39 erythromycin-resistant Staphylococcus aureus to clindamycin，penicillin and gentamicin were high，and were 100.0%，92.3% and 53.8%，respectively. A total of 8(20.5%) isolates of Staphylococcus aureus with discordant resistance pattern (erythromycin resistant and clindamycin sensitive) were all positive in D test. The positive rates of ermA，ermB and ermC were 25.6%（10/39），46.2%（18/39） and 46.2%（18/39），respectively. A total of 75.0% (6/8) and 25.0% (2/8) isolates with phenotypic expression of inducible resistance by D test harbored ermC and ermA genes. PFGE analysis of the all 10 MRSA isolates harboring ermA gene produced 6 distinct pulsotypes，8 MRSA isolates were SCCmecⅢ genotype，and 2 MRSA isolates could not be determined by SCCmec typing. Conclusions　The positive rate of erm genes in erythromycin-resistant Staphylococcus aureus in patients with bloodstream infection is relatively high. The ermA and ermB genes are mainly harbored by MRSA and methicillin-sensitive Staphylococcus aureus(MSSA)，respectively. The isolates with erythromycin inducible to clindamycin resistance mainly harbored ermC gene. A total of 10 isolates harboring ermA gene are all MRSA ，and are multidrug-resistant .There is scattered cloning spread of MRSA belonging to SCCmecⅢ genotype with ermA gene.

Analysis on the drug resistance and homology of pandrug-resistant Acinetobacter baumannii

ZHAO Fuju 1，LIU Huayong 2，ZHOU Lifang 1，FANG Yi 1，PANG Lifeng 1，LIU Wenjian 1，ZHAO Hu 1

2013, 28(3):  194-198.  DOI: 10.3969/j.issn.1673-8640.2013.04.006

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Objective　To investigate the drug resistance and homology of pandrug-resistant Acinetobacter baumannii (PDRAB) and analyze the clinical characteristics and outcome of its infection，so as to provide reference for the clinical reasonable medication and the prevention of the infection. Methods　The drug resistance of Acinetobacter baumannii from inpatients of the hospital was determined by automatic microbial identification system of BioMerieux VITEK-2 Compact and Kirby-Bauer method. Retrospective investigation was performed on the clinical characteristics and therapeutic efficacy turnover of the patients with PDRAB infection. A semiquantitative plate assay was used to test the biofilm-forming ability of PDRAB. The homology of PDRAB was detected by pulsed-field gel electrophoresis. Results　A total of 138 Acinetobacter baumannii were isolated，among which 11 (7.97%) isolates were PDRAB. A total of 10 in patients with PDRAB infection were enrolled in the study. The PDRAB infection occurred most frequently in respiratory intensive care unit，7 (70%) isolates were from respiratory intensive care unit，2 (20%) isolates were from surgical unit，and 1 (10%) isolate was from intensive care unit. The polymyxin sensitivity of all 10 PDRAB was 100%，but they were resistant to all other antibiotics in routine clinical use，and the resistance rate was 100%. There were 7 (5.07%) biofilm-positive isolates among the 138 Acinetobacter baumannii，2 isolates were PDRAB，and 5 isolates were non-PDRAB. All of 10 PDRAB were classified into 7 pulsotypes（A，B，C，D，E，F and G），the homology of 2 isolates of Pulsotype D and Pulsotype E was high，and the others were sporadic isolates. Conclusions　Pandrug-resistance is serious in respiratory intensive care unit of the hospital. Pandrug-resistance is closely related to long-term use of broad-spectrum antibiotics，invasive treatment，and patients with multiple underlying diseases. A variety of antimicrobial agents used in combination use have a certain effect on the control of PDRAB infection. It is necessary to strengthen the monitoring of drug resistant and biofilm-positive isolates，especially in respiratory intensive care unit.

A study of plasmid-mediated aac(6′)-Ib gene determination and quinolone resistance

ZHAO Qian，WANG Yaping，YING Chunmei，ZHENG Bing，ZHANG Haomin，YANG Jun

2013, 28(3):  199-202.  DOI: 10.3969/j.issn.1673-8640.2013.03.007

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Objective　To investigate the quinolone resistance to Escherichia coli isolates，and to identify the relationship of existence of plasmid-mediated aac (6′)-Ib genes with the resistance of quinolones.  Methods　Kirby-Bauer(k-B) method was performed for 121 Escherichia coli isolates from urine samples collected from March 2010 to March 2011 in Renji Hospital. The aac (6′)-Ib genes in Escherichia coli isolates were determined by polymerase chain reaction(PCR). The amplification fragment of positive isolates were selected to be sequenced，and the genotypes were determined.  Results　Escherichia coli isolates′ resistance rates of nalidixic acid，ciprofloxacin，norfloxacin and levofloxacin were all higher than 75%. The determination rate of aac(6′)-Ib gene was 14.0% (17/121)，and the mutation rate was 82.4%(14/17). The aac (6′)-Ib gene positive isolates had higher resistance rates of ampicillin-sulbactam，piperacillin-tazobactam，cefazolin，cefaclor，cefuroxime，amikacin，ciprofloxacin，norfloxacin and levofloxacin than the negative isolates. There was statistical significance(P<0.05).  Conclusions　Quinolone resistance to Escherichia coli isolates in Renji Hospital is serious. With the plasmid-mediated aac (6′)-Ib genes，the Escherichia coli isolates may have an increasing resistance rates to quinolones and decreasing susceptibility to beta-lactamase inhibitors and cephalosporins antibiotics.

Investigation on the reference intervals of blood cadmium and urine mercury in Jiangsu，Zhejiang，Shanghai and Anhui regions among 18-22-year-old males

PAN Chang 1，LIN Weidong 1，ZHOU Gang 1，CHEN Xiangfang 2

2013, 28(3):  203-206.  DOI: 10.3969/j.issn.1673-8640.2013.03.008

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Objective　To investigate the reference intervals of blood cadmium and urine mercury in Jiangsu，Zhejiang，Shanghai and Anhui regions among 18-22-year-old males，and to provide realistic reference standards for firefighter occupational health examination of heavy metal screening.  Methods　The atomic absorption spectrophotometry was used for blood cadmium determination in 1 683 new firefighters. The acidic stannous chloride reduction method was used for the determination of urine mercury，and the results were calibrated by creatinine（Cr）. The blood cadmium and urine mercury values were analyzed statistically. The results were classified and analyzed comparatively according to regions and smoking with reference intervals calculated by P₀-P₉₅.  Results　In Jiangsu，Zhejiang，Shanghai and Anhui regions among 18-22-year-old males，the blood cadmium values were positively skewed distribution，and the reference interval was 0-6.70 μg/L. Urine mercury values were positively skewed distribution，and the reference interval was 0-3.00 μg/g Cr. The regional comparison had no statistical significance（P＞0.05）. The comparison between smoking group and non-smoking group had statistical significance（P＜0.05）.  Conclusions　The reference intervals of blood cadmium and urine mercury should be established and classified between different regions and different ages.

Changes of urine conductivity in patients with urinary stones

ZENG Tingting，HUANG Yuxia，WANG Dengchao，HUANG Qian，CHEN Jiao，SU Jun

2013, 28(3):  207-210.  DOI: Department of Clinical Laboratory，Huaxi Hospital，Sichuan University，Sichuan Chengdu 610041，China

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Objective　To investigate the changes of urine conductivity in healthy subjects and patients with urinary stones and the clinical application of urine conductivity.  Methods　A total of 436 healthy subjects and 348 patients with kidney and ureter stones diagnosed by ultrasound were enrolled. UF1000i automatic urine sediment analyzer was used to determine urine conductivity.  Results　Random urine conductivity showed normal distribution in healthy subjects. Urine conductivities of healthy subjects who were older than 40 years old were lower than those of subjects who were 40 years old or younger (P＜ 0.001). Urine conductivities of patients with kidney and ureter stones were lower than those of healthy subjects (P＜ 0.001). There was no difference in urine conductivities between patients with different-size stones. Receiver operating characteristic (ROC) curves were drawn by making urine conductivity as a diagnostic test of stone disease，and the areas under ROC curves were＜0.3.  Conclusions　Urine conductivities in patients with urinary stones are lower than those in healthy subjects，and the size of urinary stone does not influence the urine conductivity. Urine conductivity can not be used as a diagnostic test of urinary stones，but can be used as a screening experiment to remind doctors doing ultrasound to detect urinary stones combined with the other urine parameters.

Application evaluation of liquid MGIT culture method in detection of Mycobacterium tuberculosis

WU Xuebing 1，LU Bin 2，GUI Xiaohong 3，JIANG Yuan 3，ZHANG Guoying 2，WANG Hong 2

2013, 28(3):  211-214.  DOI: 10.3969/j.issn.1673-8640.2013.03.011

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Objective　To use liquid Mycobacterium growth indicator tube (MGIT) culture method，solid Roche(L-J) culture method and sputum direct smear acid-fast staining test for the detection of Mycobacterium tuberculosis，and to evaluate the application of liquid MGIT culture method in the diagnosis of tuberculosis.  Methods　From November 2010 to August 2011 in Shanghai Songjiang District Central Hospital，1 598 sputum samples were collected from the suspected and confirmed tuberculosis inpatients and outpatients，who were diagnosed again after treatment (828 suspected newly cases and 770 cases from the follow-up patients being diagnosed again)，and all cases were detected by liquid MGIT culture method，solid L-J culture method and sputum direct smear acid-fast staining test.  Results　By liquid MGIT culture method，the reporting positive time of suspected newly cases was (13.63±7.14) d，and the reporting positive time by solid L-J culture method was (28.67±10.04) d，with statistical significance (P＜0.05) . The reporting positive time of the being diagnosed again cases by liquid MGIT culture method was (22.94±9.55) d，and was (28.53±10.40) d by solid L-J culture method with statistical significance (P＜0.05 ). The positive rate of suspected newly patients by liquid MGIT culture method was 32.00% (265/828)，which was higher than those by sputum direct smear acid-fast staining test [14.80% (123/828)] and solid L-J culture method [17.90% (148/828)] with statistical significance (P＜0.05). The positive rate of being diagnosed again cases by liquid MGIT culture method was 9.30% (72/770) ，and was 6.10% (47/770) by sputum direct smear acid-fast staining test without statistical significance (P＞0.05 )，but that by liquid MGIT culture method was higher than that by solid L-J culture method [2.60% (20/770)] (P＜ 0.05). The contamination rate of liquid MGIT culture method [6.57% (105/1 598)] was higher than that by solid L-J method [4.00% (64/1 598)] (P＜ 0.05).  Conclusions　The liquid MGIT culture method in terms of increasing the positive rate and shortening the reporting positive time is better than traditional detection methods，and it is easy to operate without using expensive equipment and suitable to use in the primary hospital.

The intracellular localization and clinical significance of HNF4α in tissues of colorectal cancer

ZHANG Liang 1，YUAN Huixiong 1，HUANG Yongzhi 2

2013, 28(3):  215-217.  DOI: 10.3969/j.issn.1673-8640.2013.03.012

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Objective　To detect the intracellular localization and expression of hepatocyte nuclear factor-4 alpha (HNF4α) in tissues of colorectal cancer，and to investigate its relationship with clinical pathological parameters in patients with colorectal cancer.  Methods　Immunohistochemistry was used to detect the intracellular localization and expression of HNF4α in 132 specimens of colorectal cancer tissues and adjacent normal intestinal mucosa.  Results　The intracellular localization of HNF4α in colorectal cancer tissues was located in the cell plasma or in the cell nuclear. The positive rate of HNF4α expression in the cell plasma was significantly higher in colorectal cancer tissues than that in adjacent normal intestinal mucosa (76.5% and 3.8%，P＜0.05). The positive rate of HNF4α expression in the cell nuclear was significantly lower in colorectal cancer tissues than that in adjacent normal intestinal mucosa (12.9% and 64.4%，P＜0.05). The expression of HNF4α in the cell plasma of colorectal cancer tissues was related to tissue differentiation degree and Dukes staging(P＝0.02，P＝0.03)，but not to sex，age，tumor location，or lymph node metastasis(P＞0.05).  Conclusions　The high expression of HNF4α in the cell plasma or low expression in the cell nuclear of colorectal cancer tissues is related with the development of colorectal cancer. The abnormal localization of HNF4α in colorectal cancer can serve as an objective index for the diagnosis and prognosis of colorectal cancer.

Significance investigation of neutralization confirmatory test for the weak reactive result of HBsAg determination by electrochemiluminescence immunoassay

HU Yao，LIU Weiwei，HUANG Zhiji

2013, 28(3):  218-220.  DOI: 10.3969/j.issn.1673-8640.2013.03.013

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Objective　To use neutralization confirmatory test to confirm and analyze the results of weak reactive hepatitis B surface antigen (HBsAg) determination by electrochemiluminescence immunoassay (ECLIA).  Methods　A total of 100 weak reactive HBsAg positive samples (1.00＜ COI ＜50.00) were detected by ECLIA，and confirmed by neutralization confirmatory test. The results were analyzed.  Results　Among the 100 weak reacitve HBsAg positive samples，87 samples (87.0％) were confirmed，10 samples (10％) were negative，and 3 samples (3.0％) were uncertain．According to the receiver operating characteristic (ROC) curve，when the COI was 1.97，the specificity was 100.0％，and the sensitivity was 83.9％. Conclusions　The neutralization confirmatory test can be used as the further means to confirm the samples with weak reactive HBsAg. ECLIA shows high sensitivity and specificity on weak reactive HBsAg positive samples．When COI＞2.00，generally it is unnecessary to do the neutralization confirmatory test and could exclude the possibility of false-positivity

Rapid detection of Streptococcus pneumoniae by real-time fluorescence quantitation PCR

YAN Shanhuo，SUN Lei，FU Kepeng，ZHUO Yongguang，YANG Shanye，FAN Zuqian，HUANG Yongxia

2013, 28(3):  221-224.  DOI: 10.3969/j.issn.1673-8640.2013.03.014

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Objective　To establish real-time fluorescence quantitation polymerase chain reaction (PCR) for the detection and epidemiological studies of Streptococcus pneumoniae.  Methods　The autolysin (lyt) gene and hemolysin (ply) gene sequences were selected to develop primers and probe. A total of 10 strains of Streptococcus pneumoniae，13 strains of non-Streptococcus pneumoniae DNA and the different concentration strains of Streptococcus pneumoniae DNA were detected by real-time fluorescence quantitation PCR，and 200 clinical specimens were detected by primers and probe. The specificity and sensitivity were also analyzed.  Results　The 10 strains of Streptococcus pneumoniae were measured to obtain amplification products. However，13 strains of non-Streptococcus pneumoniae DNA had no obvious amplification，and the sensitivity was 100fg. Among the 200 clinical specimens，real-time fluorescence quantitation PCR detected 42 cases of Streptococcus pneumoniae-positive (the positive rate was 21%)，while the culture method detected 16 cases of Streptococcus pneumoniae-positive (the positive rate was 8%).  Conclusions　Real-time fluorescence quantitation PCR is a rapid，sensitive and specific assay for the detection of Streptococcus pneumoniae. It can be used for the diagnosis and epidemiological studies.

Research on the lipoprotein and its subclass determination by nanotechnology-based microchip capillary electrophoresis

WANG Hua 1，HAN Chongxu 2，WANG Huimin 1，JIN Qinghui 3，WANG Daxin 1，CAO Li 1，DONG Lanmei 1

2013, 28(3):  225-228.  DOI: 10.3969/j.issn.1673-8640.2013.03.015

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Objective　To establish a method for separating serum lipoprotein and its subclass by microchip capillary electrophoresis，and investigate the significance of clinical application.  Methods　The pre-stained lipoproteins with NBD C6-ceramide were separated by Tricine buffers with gold nanoparticles as an additive in quartz chip with laser induced fluorescence detector. The impact of gold nanoparticle with different sizes and concentrations on the separation of lipoprotein was investigated. The best electrophoresis condition was obtained. The detection limit，linearity and reproducibility were investigated. Serum lipoproteins of 30 patients with coronary heart disease (CHD) and 30 healthy subjects (control group) were separated by microchip capillary electrophoresis.  Results　The 20 nmol/L 5nm gold nanoparticles were used as an additive to the buffer in order to obtain the absolute separation of lipoproteins [large and buoyant low density lipoprotein (LDL) (lLDL)，small and dense LDL (sdLDL)，very low-density lipoprotein (VLDL) and high-density lipoprotein(HDL)]. Under optimized conditions，the linear ranges of lLDL，sdLDL，VLDL and HDL were 0.01-0.8，0.04-1.0，0.04-1.0 and 0.02-0.8 mg/L，and their limits of detection were 5，5，15 and 8 μg /L，respectively [signal-to-noise ratio (S/N) ＝ 3]. Relative standard deviation (RSD) values of the peak areas of lLDL，sdLDL，VLDL and HDL were 3.5%，2.2%，4.5% and 3.9%，respectively. The levels of sdLDL and VLDL increased significantly，and HDL decreased more significantly in CHD group than in control group.  Conclusions　The results show that nanotechnology-based microchip capillary electrophoresis is applicable for the rapid and convenient detection of lipoprotein and its subclass，and it has significance for evaluating the analysis of CHD risk factors and the early detection and prevention of CHD.

Comparison on the consistency of two electrophoresis methods in the detection of hemoglobin

CHEN Yongxiu 1，ZHOU Yunzhen 2，CHEN Jinhong 3，LI Shuquan 4，LONG Guifang 4

2013, 28(3):  229-232.  DOI: 10.3969/j.issn.1673-8640.2013.03.016

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Objective　To compare the results′consistency of agarose gel electrophoresis and cellulose acetate membrane electrophoresis in the detection of hemoglobin.  Methods　By Sebia Hydrasys LC automated agarose gel electrophoresis and cellulose acetate membrance electrophoresis，the hemoglobin levels of 4 126 patients with Mediterranean anaemia were detected（pH value ＝8.6）. The HbA₂，HbH，and HbBart′s contents were determined by spectrophotometry. The HbF content was determined by alkaline denaturation test. The results of the 2 methods were compared. The cases with different results by the 2 methods were confirmed by genetic analysis.  Results　A total of 4 126 cases by agarose gel electrophoresis had 686 cases of beta-Mediterranean anaemia （HbA₂ and/or HbF increasing）and 321 cases of alpha-Mediterranean anaemia（HbA₂ decreasing）. Cellulose acetate membrane electrophoresis had 683 cases of beta-Mediterranean anaemia and 323 cases of alpha-Mediterranean anaemia. Basically，the results were consistent forHbA₂ and HbF by the 2 methods. Agarose gel electrophoresis detected 103 cases of HbH，71 cases of HbBart′s and 63 cases of HbCS. Cellulose acetate membrane electrophoresis detected 141 cases of HbH，101 cases of HbBart′s and 65 cases of HbCS. After being confirmed by genetic analysis，agarose gel electrophoresis missed 38 cases of HbH，30 cases of HbBart′s and 2 cases of HbCS.  Conclusions　Agarose gel electrophoresis can accurately detect HbA2 and HbF，and it has deficiencies for HbH and HbBart′s. While HbA₂ decreasing，isopropyl alcohol-test positive and erythrocyte fragility test （one tube method） positive，cellulose acetate membrane electrophoresis should be used to the quantitive determination of HbH and HbBart′s. Electrophoresis can be used in the screening and general investigation of Mediterranean anaemina，and genetic analysis should be used in clinical confirmation and antepartum diagnosis.