Table of Content

15 May 2013, Volume 28 Issue 5

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The metallo-beta-lactamase detection and antibiotic resistance analysis of imipenem-resistant Pseudomonas aeruginosa

HE Lizhi，HUANG Luping，LI Zhifang，PENG Aihong

2013, 28(5):  353-356.  DOI: 10.3969/j.issn.1673-8640.2013.05.001

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Objective　To analyze the antibiotic susceptibility of metallo-beta-lactamase (MBL) of imipenem- resistant Pseudomonas aeruginosa(PA)，and provide the reference for nosocomial infection prevention. Methods　A total of 460 PA were isolated and collected from 2009 to 2011，and the imipenem-resistant isolates were screened. With the combination of disk diffusion method，the MBL phenotype of imipenem-resistant isolates was detected. The MBL genotype was detected by polymerase chain reaction (PCR). The positive isolates amplified by PCR were sequenced. The antibiotic resistance of MBL-producing isolates of imipenem-resistant PA was compared with that of non MBL-producing isolates. The antibiotic resistance characteristics of the different genotypes of MBL-producing isolates were analyzed. Results　In 460 PA，28 MBL-producing isolates were identified. The 19 isolates were IMP-1 type，and 2 isolates were VIM-2 type by PCR. The 1 isolate of IMP-1 type and 1 isolate of VIM-2 type were selected randomly and sequenced. GenBank accessions were No. AY1686359 and AF191564，respectively. The resistance rate to meropenem of MBL-producing isolates was 95.2%，which was higher than that of non MBL-producing isolates(73.7%， P＜0.05). The resistance rates to other antibiotics of MBL-producing isolates were higher than those of non MBL-producing isolates，but the difference was not statistically significant (P＞0.05). The genotypes of MBL-producing isolates in imipenem-resistant PA were different，and the antibiotic resistances were also different. Conclusions　The main genotype of MBL-producing isolates in PA is IMP-1 type. These isolates have strong antibiotic resistance to a lot of antibiotics commonly used in clinic. The epidemiology surveillance of the MBL-producing isolates must be strengthened to prevent and control the resistance spread. Simultaneously，clinical antibiotics should be used according to the antibiotic susceptibility tests.

Genotyping of staphylococcal cassette chromosome mec and antimicrobial resistance of 65 isolates of methicillin-resistant Staphylococcus aureus

WANG Ailing 1 ， JI Bing 1 ， SUN Wanju 2 ，MENG Wei 1， AN Xinye 1， SU Zhenguo 1， WANG Lianwen 1， ZHANG Yumei 2

2013, 28(5):  357-361.  DOI: 10.3969/j.issn.1673-8640.2013.05.003

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Objective　To investigate the genotype distribution characteristics of staphylococcal cassette chromosome mec(SCCmec) and antimicrobial resistance of methicillin-resistant Staphylococcus aureus(MRSA). Methods　Multiplex polymerase chain reaction(PCR) was used to detect SCCmec genotype. Four antimicrobial resistant genes were amplified by traditional PCR. Antimicrobial susceptibility of 19 antibiotics was detected by disk diffusion method. Results　The 60 isolates of MRSA were SCCmec Ⅲ， only 4 isolates were SCCmec Ⅳa， and 1 isolate had no genotype.  All of the 65 isolates of MRSA were sensitive to linezolid， quinupristin/dalfopristin， vancomycin and teicoplanin， and were low resistant to chloramphenicol (4.62%). The positive rates of aac(6′)/aph(2′′)， aph(3′)Ⅲ， tetM and ant(4′) were 93.85%，52.31%，92.31% and 9.23%. Conclusions　The main SCCmec genotype of MRSA is SCCmecⅢ. MRSA has multiple resistances to commonly used antibiotics．There are high percentages of aminoglycoside and tetracycline．The multiplex PCR can be applied into genotyping study of MRSA SCCmec effectively.

Comparison study on the determination of colony morphology of Ureaplasma urealyticum by two kinds of culture media

ZHOU Yunheng 1，MA Hongxia 2， CAO Guangya 1，SHI Xiaoxing 1，LIU Fenghua 1

2013, 28(5):  362-365.  DOI: 10.3969/j.issn.1673-8640.2013.05.004

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Objective The results of detecting Ureaplasma urealyticum(Uu) from different kinds of samples in the liquid medium and the selective solid medium were compared, and morphological characters of Uu colonies by the selective solid culture medium were observed. Methods The 1698 samples from urogenital tract were cultured in the liquid medium and the selective solid medium. The positive rates from the two assays and three kinds of samples were compared and analyzed. The colonies features of Uu were observed by the selective solid medium. Results The positive rate of Uu from female urogenital tract was significantly（49.9%）higher than that from semen（39.6%）and urethral discharge（19.2%）（P<0.05）. There was significant difference of the positive rate from female urogenital tract between the liquid medium and the selective solid medium（P<0.05）. However, there were no statistics significance of the positive rate from male urogenital tract between the two assays（P>0.05）. The main colonies of Uu was typical echinus colony(77.5%), fried egg colony（13.8%）, vesicle colony, tiny colony, coryneform colony and mixed colony. Conclusions The positive rate of Uu from female urogenital tract is higher than that from semen samples, and the positive rate from male urethral discharge is the lowest. The results of Uu from female specimens using two media are different obviously. There is a good consistency between the two ways about the male samples. Uu colonies have morphological heterogeneity.

Study of hyperinsulinism progression in patients with type 2 diabetes mellitus

LI Sha 1，LIU Rui 2，WU Yiqing 2，WU Lina 1

2013, 28(5):  366-369.  DOI: 10.3969/j.issn.1673-8640.2013.05.005

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Objective　To investigate the progression of hyperinsulinism(HINS) in different development phases of type 2 diabetes mellitus （T2DM）.  Methods　A total of 38 pre-T2DM patients，52 newly diagnosed T2DM patients and 92 patients who had T2DM history were enrolled. According to the course of T2DM，the patients with T2DM history were classified into 3 groups：0.5-3 years，4-7 years and 8-11 years， and 20 healthy subjects were as controls. The glycosylated hemoglobin (HbA1c)，blood glucose，insulin and C peptide were determined．Homeostasis model of insulin resistance index（HOMA2-IR） and secretion function index (HOMA2-%β) were calculated by HOMA2 calculator (V2.2) downloaded from www.ocdem. ox.ac.uk.  Results　All T2DM groups′ secretion peaks appeared in 2 h of oral glucose tolerance test(OGTT)， and all presented secretion delay. The 2 h insulin levels of the pre-T2DM and newly diagnosed T2DM groups were higher than HINS diagnosis standard，and the HINS patients accounted for 50.00% and 48.10%， respectively. With the increasing of T2DM course，the proportions of HINS patients in the T2DM history group were 28.57%，13.89% and 10.00%， respectively.  Conclusions　There is a proportion of pre-T2DM patients with HINS，and with the increasing of T2DM course，the proportion of HINS patients reduces，but HINS still exists in patients with long history of T2DM.

The relationship of immunoglobulins， complements， albumin， blood lipids with kidney pathology in patients with nephrosis

WU Chunlin

2013, 28(5):  370-373.  DOI: 10.3969/j.issn.1673-8640.2013.05.006

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Objective　To investigate the relationship of serum immunoglobulins （IgG， IgA）， complements（C3， C4）， albumin（Alb）， 24 h urinary protein， blood lipids[total cholesterol(TC) and triglyceride(TG)] with kidney pathology in patients with nephrosis.  Methods　A total of 251 patients with nephrosis were enrolled. The correlations of serum IgG， IgA， C3， C4 and glomerular mesangial cell proliferation score with Alb， 24 h urinary protein， TC， TG， and the scores of IgG， IgA and C3 immune fluorescence intensities were analyzed. There were IgA nephropathy（IgAN， 106 cases）， membranous nephropathy（MN， 19 cases）， focal segmental glomerulosclerosis（FSGS， 14 cases）， minimal change disease (MCD， 7 cases)， lupus nephritis（LN， 7 cases）， hepatitis B virus-associated glomerulonephritis（HBV-GN， 5 cases）， hypertensive renal damage（8 cases）and healthy controls（56 cases）. Their levels of IgG， IgA， C3 and C4 were compared. Results　Serum IgG， IgA and Alb were positively correlated with IgA and C3 immune fluorescence intensities（P＜0.05）. Serum C3 and C4 had positive correlation with TC and TG（P＜0.05）， and had negative correlation with IgG， IgA and C3 （P＜0.05）. The score of glomerular mesangial cell proliferation with serum IgA， TG and the immune fluorescence intensities of IgA and C3 showed a positive correlation（P＜0.05）， and it with serum C3 showed a negative correlation（P＜0.05）. Serum IgG level of MCD group was the lowest， and the comparison of MCD group with IgAN， FSGS， LN， HBV-GN， hypertensive renal damage and healthy control groups was statistically significant（P＜0.05）， but the comparison of MCD group with MN group was not statistically significant（P＞0.05）. Serum IgA level of IgAN group was the highest， and the comparison of IgAN group with MN， FSGS， MCD and healthy control groups was statistically significant（P＜0.05）， but the comparison of IgAN group with LN， HBV-GN， hypertensive renal damage groups was not statistically significant（P＞0.05）. Serum C3 and C4 levels of LN group was the lowest， and the comparison of LN group with the other groups was statistically significant（P＜0.01）. Conclusions　Immunoglobulins， complements， urinary protein， Alb， blood lipids for the diagnosis， treatment and prognosis assessment of kidney disease have important guiding significance. However， it has a lot of influencing factors， and the diagnosis and treatment of kidney disease can not only rely on the changes of one or several index levels， but also should refer to whether there are influencing factors， combined with more clinical indices and kidney pathology in making comprehensive judgments.

Clinical significance of urinary NGAL determination in tumor patients with renal tubular injury by cisplatin chemotherapy

HU Leiguang 1，ZHU Liyue 1，XIE Fang 2，CHEN Jian 1，WANG Xinlei 1

2013, 28(5):  374-378.  DOI: 10.3969/j.issn.1673-8640.2013.05.007

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Objective　To investigate the application significance of urinary neutrophil gelatinase-associated lipocalin (NGAL) determination in tumor patients with renal tubular injury by chemotherapy. Methods　The random urine specimens were collected within 24 h before and after cisplatin(DDP) chemotherapy in 58 tumor inpatients (including lung， breast， cervical， parotid gland cancers and so on) who were treated by DDP chemotherapy. There were 57 urine specimens that were collected from patients before chemotherapy. They were classified into 2 groups， 25 specimens for the patients without chemotherapy and 32 specimens for the patients with once chemotherapy at least before admission. A total of 53 of 54 urine specimens after chemotherapy paired with ones before chemotherapy. A total of 51 healthy subjects were enrolled as control group. The levels of urinary NGAL， N-acetyl-beta-D-amino glucosidase (NAG)， alpha1-microglobulin (α1-MG)， urinary microalbumin (mAlb) and urinary creatinine (Cr) were determined. According to urinary Cr(UCr)， the 4 indicators were corrected by calculating for NGAL/UCr， NAG/UCr， α1-MG/UCr and mAlb/UCr ratio. According to the NAG positive as renal tubular injury judgment index，the all objectives were classified into 2 groups of injury and non-injury. The accuracy of renal tubular injury judged by urinary NGAL/UCr was assessed by the receiver operating characteristic (ROC)curve.  Results　Comparing the results of patients with chemotherapy， patients without chemotherapy before admission and controls， there was no statistical significance (P＞0.05)， except NGAL/UCr and NAG/UCr of patients without chemotherapy were higher than those of controls (P＜0.05). Of the results between patients before chemotherapy and after chemotherapy， when the median was 25%-75%， NGAL/UCr were 1.37(0.76-2.16)mg/mmol and 2.14(1.39- 4.08)mg/mmol， NAG/UCr were 1.30(0.76-2.56) U/mmol and 2.58(1.75-4.02)U/mmol，and α1-MG/UCr were 0.21(0.04-0.74)mg/mmol and 1.00(0.30-1.91)mg/mmol(P＜0.01). The difference of mAlb/UCr was no statistically significant before and after chemotherapy(P＞0.05). Comparing NGAL/UCr abnormality with NAG/UCr， α1-MG/UCr and the combined result abnormalities of the 3 indicators， the differences of their abnormalities in patients before chemotherapy were not statistically significant (P＞0.05). The NGAL/UCr abnormality and the combined result abnormalities of the 3 indicators were higher than those of NAG/UCr and α1-MG/UCr in patients after chemotherapy (P＜0.05). The area under the ROC curve (AUC) of NGAL/UCr was 0.770 1， its 95% confidence interval was 0.698 4-0.841 7. The diagnosis cut-off value was 1.15 mg/mol (the sensitivity was 82.09%， and the specificity was 61.05%).  Conclusions　Urinary NGAL/UCr have clinical application significance in patients with renal tubular injury and kidney function decline by DDP chemotherapy. To prevent from drug nephrotoxic injury， urinary NGAL determination may become a kind of novel selectable biological marker.

Evaluation on the application of the combination determination of Hcy and BNP in acute myocardial infarction

LIU Xiaofeng，CHEN Xueli，TU Yan

2013, 28(5):  379-381.  DOI: 10.3969/j.issn.1673-8640.2013.05.008

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Objective　To investigate the application of the combination determination of plasma homocysteine (Hcy) and B-type natriuretic peptide (BNP) in acute myocardial infarction (AMI). Methods　A total of 150 patients with AMI and 135 healthy subjects were enrolled. The levels of plasma BNP and Hcy were determined after admission and drug thrombolysis or percutaneous coronary intervention (PCI) in patients with AMI. The levels of plasma BNP and Hcy were compared between 51 patients with cardiovascular events and 99 patients without cardiovascular events during hospitalization and follow-up.  Results　The levels of plasma BNP[(594.38±167.58)pg/mL] and Hcy[(37.6±11.44)μmol/L] in patients with AMI were higher than those in healthy controls[(56.32±28.79)pg/mL and (7.62±3.18)μmol/L](P＜0.05). After admission and drug thrombolysis or PCI， the levels of plasma BNP[(256.94±118.55) pg/mL or (78.03±11.24)pg/mL] and Hcy[(16.47±7.24)μmol/L or (10.33±6.09)μmol/L] in patients with AMI were lower than those before admission and drug thrombolysis or PCI[(494.37±183.63)pg/mL and  (35.53±8.12)μmol/L](P＜0.05).The levels of patients with cardiovascular events[BNP(609.15±287.37)pg/mL and Hcy (39.5±12.61) μmol/L] were higher than those without cardiovascular events[BNP(326.85±134.24)pg/mLand Hcy (14.56±7.12)μmol/mL](P＜0.05). Conclusions　The plasma BNP and Hcy are related to AMI. Plasma BNP and Hcy may be as application parameters in AMI.

Investigation on the significance of des-gamma-carboxy-prothrombin in the diagnosis of primary hepatocellular carcinoma

ZHONG Zhimin，WANG Wei，MO Yundan，DAI Hui

2013, 28(5):  382-386.  DOI: 10.3969/j.issn.1673-8640.2013.05.009

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Objective　To investigate the diagnosis significance of des-gamma-carboxy-prothrombin (DCP) in patients with primary hepatocellular carcinoma，and to improve the sensitivity and specificity in the diagnosis of primary hepatocellular carcinoma.  Methods　A total of 116 patients with primary hepatocellular carcinoma and 57 patients with secondary liver cancer，52 patients with digestive system tumors，50 patients with chronic liver disease，73 patients with other liver diseases and 50 healthy subjects (healthy control group) were enrolled. Their serum DCP and alpha fetoprotein(AFP) levels were determined by chemiluminescence. The receiver operating characteristic (ROC) curve was performed. The best diagnosis cut-off value of primary hepatocellular carcinoma was calculated.  Results　The DCP level in primary hepatocellular carcinoma group was significantly higher than those in the other groups (P＜0.05)，and there was no statistical significance among the other 5 groups （P＞0.05）. ROC curve showed that the most suitable diagnosis cut-off value of primary hepatocellular carcinoma was 44.5 mAU/mL. The sensitivity was 74.2% ， and the specificity was 88.6%. The specificity of the combination determination of DCP and AFP was above 96%， and its sensitivity was above 91%. There was no correlation between DCP and AFP (r=0.026，P=0.782).  Conclusions　DCP has diagnosis significance in the diagnosis of primary hepatocellular carcinoma，and in primary hepatocellular carcinoma， secondary liver cancer and other benign liver diseases ， it has identification significance. The combination determination of DCP and AFP can improve the diagnosis sensitivity and specificity of primary hepatocellular carcinoma.

The correlation analysis of traditional Chinese medicine syndrome with pathological diagnosis，TNM stage and tumor markers in patients with lung cancer

WANG Lei 1，NING Xiaoxiao 1，LI Hegen 2， WANG Qihua 1，XU Weijie 2， ZHOU Lei 2， XU lin 2

2013, 28(5):  387-390.  DOI: 10.3969/j.issn.1673-8640.2013.05.010

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Objective　To investigate the coorelation of pathological diagnosis，TNM stage and the levels of tumor markers with different scenarios of traditional Chinese medicine syndrome in patients with lung cancer. Methods　A total of 196 patients had been diagnosed with lung cancer. The correlation analysis of pathological diagnosis[non-small cell lung cancer(squamous carcinoma， adenocarcinoma， large cell carcinoma and mixed cell carcinoma) and small cell lung cancer]，TNM stage and tumor markers[cytokerantin-19-fragment(CYFRA21-1)，carcinoembryonic antigen (CEA)，carbohydrate antigen 125(CA125)，pro-gastrin-releasing peptide(ProGRP) and squamous cell carcinoma antigen(SCC Ag)] with different scenarios of traditional Chinese medicine syndrome in patients with lung cancer was performed.  Results　Most of the traditional Chinese medicine syndromes in lung cancer patients were lung and spleen deficiency syndrome(119 cases， 60.71%)， Qi and Yin deficiency syndrome(76 cases， 38.7%) and spleen and kidney deficiency syndrome(1 case， 0.52%). However，the distribution of the traditional Chinese medicine syndrome among the different pathological diagnosis in lung cancer patients showed no significant difference （P=0.325）. Lung and spleen deficiency syndrome was concentrated in TNM stage Ⅰ-Ⅲ patients，while Qi and Yin deficiency syndrome was concentrated in TNM stage Ⅳ patients. The CYFRA21-1 levels in Qi and Yin deficiency syndrome patients were higher than those in lung and spleen deficiency syndrome patients（P=0.017）. The differences of CEA，CA125，ProGRP and SCC Ag levels were not significant （P＞0.05）. Conclusions　Most of the lung cancer patients are deficiency syndrome. It indicates that advanced stage lung cancer patients would be in unstable conditions and have bad prognosis when Qi and Yin deficiency. CYFRA21-1 and other tumor markers could be the microscopic evidence to traditional Chinese medicine syndrome diagnosis.

The significance of IGF-1 and IGFBP-3 in the diagnosis of gastric cancer

PENG Chong，SUN Guirong，ZHANG Nana，GAO Zhenliang，Lü Xiaoyu

2013, 28(5):  391-395.  DOI: 10.3969/j.issn.1673-8640.2013.05.011

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Objective　To evaluate the clinical significance of serum insulin-like growth factor-1(IGF-1) and insulin-like growth factor binding protein-3(IGFBP-3) in the diagnosis of gastric cancer and gastric precancerous lesion. Methods　The serum levels of IGF-1，IGFBP-3，carcinoembryonic antigen (CEA) and carbohydrate antigen 199(CA199) were measured by chemiluminescence immunoassay，enzyme-linked immunosorbent assay(ELISA) and electro-chemiluminescence immunoassay in 90 patients with gastric cancer，40 patients with gastric benign disease and 50 healthy subjects. Results　The serum levels of IGF-1 in patients with gastric cancer were significantly higher than those in patients with gastric benign disease and healthy subjects（P＜0.05，P＜0.001）. The serum levels of IGFBP-3 in patients with gastric cancer were significantly lower than those in patients with gastric benign disease and healthy subjects(P＜0.001).The serum levels of IGF-1 in the early (Ⅰ-Ⅱ) stage of gastric cancer were lower than those in the late (Ⅲ-Ⅳ) stage， but were higher than those in the healthy subjects，and the serum levels of IGFBP-3 were higher than those in the late stage， but were lower than those in the healthy subjects.The serum levels of IGF-1 had significantly negative correlation with IGFBP-3，and the linear correlation equation was Y =-0.33X +116.65(r=-0.566，P＜0.01). There was no correlation between IGF-1 and IGFBP-3 with CEA and CA199. Using the healthy subjects as controls，the areas under the receiver operating characteristic(ROC) curve (AUC) of IGF-1  was obviously higher than those of CEA and CA199(P＜0.01，P＜0.05). The sensitivity of IGF-1 for the diagnosis of gastric cancer was higher than those of CEA and CA199(P＜0.05). Conclusions　IGF-1 is better than CEA and CA199， in the diagnosis of gastric cancer，and it is helpful for the evaluation of gastric cancer stages.

The changes and clinical significance on the expressions of Th17， Th1 and cytokines in patients with systemic lupus erythematosus

XU Weijia 1，LI Zhi 1，YANG Tingting 2 ，WANG Bo 1

2013, 28(5):  396-399.  DOI: 10.3969/j.issn.1673-8640.2013.05.012

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Objective　To investigate the expressions of T helper cell 17 (Th17) ， T helper cell 1 (Th1) and cytokines such as interleukin 17 (IL-17)， interleukin 23（IL-23） and interferon-gamma (IFN-γ) in peripheral blood from patients with systemic lupus erythematosus（SLE），and their clinical significance. Methods　According to SLE disease activity index (SLEDAI)，38 patients with SLE were classified into active group(SLEDAI≥10，20 patients) and inactive group(SLEDAI＜10，18 patients). Flow cytometry was used to analyze the percentages of Th17 and Th1 in 38 patients with SLE and 20 healthy controls. The expressions of IL-17， IL-23 and IFN-γ were determined by enzyme-linked immunosorbent assay(ELISA)，and the results were analyzed statistically. Results　The percentage of Th17 in patients with SLE was significantly higher than that in healthy controls（P＜0.05），and the percentage of Th17 in active group was higher than that in inactive group(P＜0.05)， but the expression of Th1 had no statistical significance with that of healthy controls（P＞0.05） . The expressions of IL-17， IL-23 and IFN-γ in patients with SLE were higher than those in healthy controls（P＜0.05）. The expression of Th17 and the levels of IL-17 and IL-23 were positively correlated  with SLEDAI(r=0.537， 0.428 and 0.525， P＜0.05).  Conclusions　The expression of Th17 and the levels of IL-17， IL-23 and IFN-γ increase in patients with SLE， and Th17 and Th1 may play important roles in the pathogenesis of SLE.

Detection and significance of serum cross-linked carboxyterminal telopeptide region of type Ⅰ collagen and matrix metalloproteinase enzyme-3 in patients with early rheumatoid arthritis

DONG Yiyu 1，ZHANG Xingen 2，LU Jianliang 3，YU Guofang 4，LU Xinli 1

2013, 28(5):  400-403.  DOI: 10.3969/j.issn.1673-8640.2013.05.013

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Objective　To investigate the significance of cross-linked carboxyterminal telopeptide region of type Ⅰ collagen (ICTP)， matrix metalloproteinase-3 (MMP-3)， anti-cyclic citrullinated peptide (CCP) antibody， rheumatoid factor (RF) and rheumatoid arthritis(RA) activity in the early diagnosis of RA.  Methods　The enzyme-linked immunosorbent assay(ELISA) was used to determine the serum samples of 213 patients with RA (early RA group: 92 cases and medium-and long-term RA group: 121 cases)， 43 patients with non-RA associated and immune diseases and 40 healthy controls. The serum ICTP， MMP-3 and anti-CCP antibody were determined. The serum RF was determined by immunoturbidimetry.  Results　The optimal cut-off values of ICTP， MMP-3 and anti-CCP antibody in the diagnosis of early RA were 3.21 μg/L， 67.45 ng/mL， 15.45 U/mL and 12.35 U/mL. The sensitivities were 74.0%，63.4%，52.4% and 55.3%， and the specificities were 95.5%， 81.8%， 95.5% and 90.9%. RF and anti-CCP antibody were higher in the RA group than in the healthy control group and the non-RA group (P＜0.01). ICTP， MMP-3 and RF in the non-RA group and the healthy control group had statistical significance ( P＜0.01). ICTP in early RA and non-RA  group had no statistical significance (P＞0.05). The ICTP， MMP-3， anti-CCP antibody and RF of X ray staging Phase Ⅰ had statistical significances with those staging Phase Ⅱ， Ⅲ and Ⅳ(P＜0.05， P＜0.01). The ICTP， MMP-3 and anti-CCP antibody of Phase Ⅱ with those of Phase Ⅲ and Ⅳ had statistical significance (P＜0.05， P＜0.01). The RF among Phase Ⅱ， Ⅲ and Ⅳ had no statistical significance (P＞0.05). Conclusions　The simultaneous detection of ICTP， MMP-3， anti-CCP antibody and RF can improve accuracy and reduce missed diagnosis. Dynamic combination detection can monitor effectively the progression of RA.

The comparative study of gene chip and ratio drug sensitive test in detecting Mycobacterium tuberculosis resistance to rifampin and isoniazid

OU Weizheng，LUO Kewen，WANG Yan，QIN Wan，MENG Jun，ZHANG Tingmei

2013, 28(5):  404-407.  DOI: 10.3969/j.issn.1673-8640.2013.05.014

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Objective　To study comparatively the application of gene chip and ratio drug sensitive test in detecting Mycobacterium tuberculosis（MTB） resistance to rifampin（RFP） and isoniazid（INH）， and investigate a rapid method for diagnosing multi-drug resistant tuberculosis (MDR-TB). Methods　After isolating 327 strains from Lowenstein-Jensen，the gene mutation of the 8 sites in rpoB，katG and inhA genes related with MTB resistance to RFP and INH was detected， and the drug resistance was determined. Meanwhile， the ratio drug sensitive test was used to detect MTB resistance to RFP and INH， and the coincidence rates of RFP/INN were compared. Furthermore，16 strains randomly from the samples which were not consistent with the results of RFP or/and INH were performed DNA sequencing， and then the coincidence rate with the DNA sequencing was compared.  Results　The coincidence rate of MTB resistance to RFP was 93.27%，but that to INH was 95.11%. The coincidence rate of MTB resistance to RFP and INH was 90.52%. The detection rates of the 2 methods in detecting MDR-TB were 18.65% and 18.96%， respectively. Comparing the gene chip with DNA sequencing，the coincidence rate of RFP drug resistance was 81.25%， and that of INH drug resistance was 100.00%. Comparing the ratio drug sensitive test with DNA sequencing，the coincidence rate of RFP drug resistance was 50.00%，and that of INH drug resistance was 37.50%. Conclusions　The gene chip in detecting MTB resistance to RFP and INH has high sensitivity and specificity， and has an clinical significance in rapid diagnosis of MDR-TB.

Evaluation on the bias of two detection systems for serum cystatin C

DAI Yue 1，WU Wenqing 2

2013, 28(5):  408-411.  DOI: 10.3969/j.issn.1673-8640.2013.05.015

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Objective　To compare methodologically and evaluate the bias of turbidimetric assay and particle-enhanced nephelometric assay for serum cystatin C(Cys C). Methods　According to the National Committee for Clinical Laboratory Standards (NCCLS) EP9-A2 document，40 serum samples were determined in duplicate measurement by Roche MODULAR P800 automatic biochemical analyzer and Siemens BN-Ⅱ specific protein analyzer， respectively. The data were used to check outliers，linear regression was analyzed，and the predictive bias and confidence interval（CI） were calculated. Results　The linear regression equation was Y=1.216X-0.139，r 2=0.978. When the Cys C concentrations were 0.53 mg/L and 0.95 mg/L，the acceptable biases were in the range of 95%CI of predictive bias. However， when the Cys C concentration  was 7.85 mg/L，the acceptable bias was＜the low limit of 95%CIof predictive bias. Conclusions　The results of the 2 detection systems have linear correlation. The predictive bias of measured results between the 2 detection systems is acceptable except the high concentration.

Performance evaluation of light initiated chemiluminescence system for the determination of thyroid stimulating hormone among pregnant women

QIAN Jun 1，HU Deyu 1，ZHANG Xianghui 2，SONG Fang 2，ZHAO Weiguo 2

2013, 28(5):  412-415.  DOI: 10.3969/j.issn.1673-8640.2013.05.016

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Objective　To compare the determination results of thyroid stimulating hormone(TSH) by light initiated chemiluminescence（LiCA） system and electro-chemiluminescence(ECL) system among pregnant women，and evaluate the performance of domestic LiCA TSH kit.  Methods　Referring to IVD Clinic Research Guideline and the National Academy of Clinical Biochemistry (NACB) guideline，the serum samples of 384 pregnant women were determined by LiCA and ECL systems respectively， and the results were analyzed statistically. Meanwhile， the analytical sensitivity， functional sensitivity， within-run precision， linearity， recovery and interference of LiCA TSH kit were evaluated.  Results　There were high correlations in quantitative determination of TSH between the 2 systems with no statistical significance（P＞0.05). The analytical sensitivity and functional sensitivity of LiCA TSH kit were 0.001 1 and 0.017 μIU/mL. The within-run precisions of high-level and low-level quality controls (QCH and QCL) were 1.42% and 1.55%， and the recoveries were 97.01% and 101.57%. The linearity analysis showed Y=24.58X-22.9(r=0.998， P＜0.05). 250 mg/dL hemoglobin， 500 mg/dL triglyceride and 10 mg/dL bilirubin had no interference to the determination results.  Conclusions　There is no significant difference between LiCA system and import ECL system for the quantitative determination of TSH among pregnant women. LiCA is suitable to the clinical application.

The application significance of CD64 index and WBC for the early diagnosis and prognosis of sepsis in children

WANG Wenjuan 1，WANG Hao 2，CHEN Zhe 3

2013, 28(5):  416-419.  DOI: 10.3969/j.issn.1673-8640.2013.05.017

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Objective　To investigate the significance of peripheral blood CD64 index and white blood cell （WBC ）for the early diagnosis and prognosis monitoring of sepsis in children. Methods　A total of 57 children with sepsis， 65 children with pneumonia and 36 healthy children were enrolled. The index of CD64 was determined by flow cytometry， and WBC was determined by haematology analyzer. Results　The index of CD64 and WBC in sepsis， pneumonia and healthy control groups had statistical significance (P＜0.01). The index of CD64 in the blood culture positive was significantly higher than that in the blood culture negative (P＜0.05). According to the receiver operating characteristic(ROC)curve， when the cut-off value in the diagnosis of sepsis was 2.25， the sensitivity was 88%， and the specificity was 85%. After treatment， the index of CD64 decreased significantly (P＜0.01)， but not to be the normal level， and the difference was statistically significant compared with the healthy control group (P＜0.01).  Conclusions　The index of CD64 has the clinical significance for the early diagnosis and prognosis of sepsis in children.

Research on molecular mechanisms of PMA-induced platelet GPⅠbα shedding

WANG Zhicheng 1，LUO Meihong 1，XIE Rufeng 2，ZHANG Xiaofeng 1

2013, 28(5):  420-424.  DOI: 10.3969/j.issn.1673-8640.2013.05.019

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Objective　Phorbol 12-myristate-13-acetate (PMA) could induce platelet GPⅠbα shedding，however，the molecular mechanisms are not fully elucidated. This study investigates the molecular mechanisms of PMA-induced platelet GPⅠbα shedding.  Methods　Washed platelets were obtained from anti-coagulated healthy volunteers′ whole blood by centrifugation and washing. Washed platelets were pre-incubated with protein kinase C (PKC) inhibitor，reactive oxygen species (ROS) antagonist，NAD(P)H oxidase inhibitor，mitochondrial ROS antagonist，or dimethyl sulfoxide (DMSO)，and were incubated with PMA. ROS levels were measured by flow cytometry. The production of platelet GPⅠbα shedding was detected by Western blot. The nonradioactive detection was used to measure PKC activity. Results　PMA-induced platelet GPⅠbα shedding was completely inhibited by PKC inhibitor BIM or ROS antagonist dithiothreitol(DTT)， respectively. PMA did dependently elevate ROS levels. PMA-induced ROS production was inhibited by PKC inhibitor and NAD(P)H oxidase inhibitor，but not mitochondrial ROS antagonist. BIM inhibited PMA-induced PKC activity，however，DTT did not. Conclusions　PMA-induced platelet GPⅠbα shedding could be regulated by PMA-PKC-NAD(P)H oxidase-ROS-ADAM17-GPⅠbα signaling pathway.

Analysis on the differences of surface adhesion molecule CD44 activation states in normal cells and breast tumor cells

HOU Lidan，LIU Yiwen，HE Yiqing，YANG Cuixia，DU Yan，GAO Feng

2013, 28(5):  425-429.  DOI: 10.3969/j.issn.1673-8640.2013.05.020

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Objective　To detect the expressions of surface adhesion molecule CD44 in normal cells and breast tumor cells， and analyze the differences of CD44 activation states in order to provide a new theoretical reference for CD44 targeted cancer therapy.  Methods　The expressions of CD44 in normal peripheral blood mononuclear cells（PBMCs），normal mouse fibroblast NIH3T3 cells and human breast tumor cell Hs578T and BT-549 were determined by flow cytometry. The activation states of CD44 were analyzed by flow cytometry， using fluorescence-labeled hyaluronic acid（FL-HA）， and then the targeted binding ability of HA-CD44 was observed by cell immunofluorescence. Results　CD44 was abundantly expressed in normal PBMCs， NIH3T3 and breast tumor cell Hs578T and BT-549 (the positive expression rate was ＞95%). The binding ability of the breast tumor cell surface CD44 with FL-HA (＞90%) was much stronger than those of PBMCs[(3.61±2.65)%] and NIH3T3[(14.33±1.35)%]. NIH3T3 cell showed almost no fluorescence(HA targeted binding without CD44)， while Hs578T and BT-549 showed strong fluorescence (HA targeted binding with CD44)according to cell immunofluorescence.  Conclusions　Normal cell PBMCs and NIH3T3 have high expressions of CD44 and low binding ability with HA， while breast tumor cell Hs578T and BT-549 have high expressions of CD44 and high binding ability with other HA. CD44 in normal cells and breast tumor cells shows 2 different activation states：inactive and active. The anti-tumor drugs with HA as targeted mononuclear and CD44 as target can select high-expression CD44 killer tumor cells and have no influence on normal cells.

Preparation and performance evaluation of a composite lipid quality control material

TANG Liping 1，ZHANG Ruigao 2，JU Yi 1，WANG Meijuan 1，HU Chunli 2，ZHU Peichao 2，ZHAN Fangyao 2

2013, 28(5):  430-433.  DOI: 10.3969/j.issn.1673-8640.2013.05.022

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Objective　To prepare a liquid composite lipid quality control material without assigned value， and then to investigate its stability. Methods　Pooled human serum[negative for the antibodies to human immunodeficiency virus (HIV)， hepatitis C virus (HCV) and hepatitis B surface antigen (HBsAg)] was used as the medium for the composite lipid quality control material preparation. The precision (lot to lot difference) and stabilities after opening and in storage were evaluated.  Results　The precision (lot to lot difference) of the quality control material was ＜ 1/4 of Clinical Laboratory Improvement Amendments (CLIA′88) acceptable performance range. The quality control material was stable for 18 months at -20℃， and the bias of each item conformed to the 1/2 CLIA′88 acceptable performance range. The stability[coefficient of variation (CV)] of each item of the opened quality control material for 19 d at 2-8℃ was ＜ 1/4 CLIA′88 acceptable performance range. Conclusions　The liquid composite lipid quality control material has good homogeneity and stability， and it can be used for the internal quality control (IQC)， external quality assessment (EQA) and harmonization survey.