Table of Content

10 May 2012, Volume 27 Issue 5

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HPLC determination of thiamine and its derivatives in human whole blood

FEI Guo-Qiang;LI Xiao-Ge;ZHONG Chun-Jiu

2012, 27(5):  331-335. 

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Objective　To establish a stable and rapid method for determining the levels of human whole blood thiamine and its phosphate ester derivatives by high performance liquid chromatography (HPLC).　Methods　After blood proteins were precipitated by trichloroacetic acid (TCA), thiamine and its phosphate esters were derivatized using potassium ferricyanide into thiochromes. The derivatives of thiamine and it phosphate esters were separated by a reversedphase HPLC column and measured by fluoroscope. The method was validated by the analysis of linearity, extraction ratio and repeatability. Results　The lower limits of quantification of blood thiamine diphosphate (TDP), thiamin monophosphate (TMP) and thiamine determinations were 8.0, 1.5 and 2.5nmol/L respectively. The signal to noise ratio (S/N) for determining thiamine and its derivatives was>10. The linear relationship was stable (r>0.99). The recovery rates of TDP, TMP and thiamine were 103.7%, 96.8% and 101.7% respectively. The repeatabilities were good. All the coefficients of variation (CV) were <10%. Conclusions　The HPLC with thiamine derivatization by potassium ferricyanide for determining thiamine and its phosphate ester derivatives is reliable and can be used for clinical analysis.

The Analysis of Aminoglycoside Interference in Urine Total Protein Determination

LU Yi-De;YANG 　Fan;DONG Dan-Feng;YANG Chi-Hui;PENG Yi-Bing

2012, 27(5):  336-339. 

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Objective　To investigate the practical application of the interference screening and quantification for urine analysis in clinical chemistry according to the  Clinical and Laboratory Standards Institute (CLSI) EP7-A2 document，and evaluate the interference effects of aminoglycoside antibiotic in urine total protein determination.   Methods　According to the CLSI EP7-A2 document， fresh urine specimens with the urine total protein concentrations of 77， 146， 538 and 832 mg/L were collected, and the etimicin sulfate solution of 3 times maximum therapeutic dosage per liter of urine was added. The interference was screened by pyrogallol red-molybdate assay. The interference effects were quantified, and the etimicin sulfate solution and distilled water were added to form a concentration gradient （0, 0.15, 0.30, 0.45 and 0.60 mg/mL） of the interferent for each group. By multiple regression analysis, the degrees of interference at any interferent concentrations were estimated.  Results　After adding 0.6 mg/mL of etimicin sulfate solution into 4 groups （with the urine total protein concentrations of 77，146，538 and 832 mg/L）， the lower confidence limit of interference effect exceeded the allowable error at the medical decision concentration. Adding 5 concentrations of the etimicin sulfate solution into each specimen group， the five-level dose-response series (0, 0.15, 0.30, 0.45 and 0.60 mg/mL) all showed a linear relationship, and the linear equations were Y＝202X－1.2，Y＝187.3X＋4，Y＝325.3X＋0.6 and Y＝345.3X＋6.4. It indicated that the positive interference effects caused by etimicin sulfate while adding 0.08，0.15，0.35 and 0.50 mg/mL of the interferent into each urine group, all exceeded the allowable error.  Conclusions　The etimicin sulfate causes a positive interference in the pyrogallol red-molybdate assay for urine total protein determination. Clinical laboratories need to perform the interference tests to screen potential interferents, quantify interference effects, evaluate potential hazards and establish meaningful interference claims.

Diagnostic Performances of Laboratory kidney Function Tests for Chronic Kidney Disease

WANG Min;WANG Qi;GU Jie;YUAN Wei-Xiang;XIONG Li-Fan

2012, 27(5):  340-344. 

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Objective　To analyze the diagnostic performances of laboratory kidney function tests for chronic kidney disease.  Methods　A total of 73 patients with chronic kidney disease were classified into 3 groups according to different estimated glomerular filtration rate (eGFR)：Group A（33 cases），eGFR:＞60 mL/（min·1.73 m²）；Group B（32 cases），eGFR: 30-60 mL/（min·1.73 m²）；Group C（8 cases），eGFR:＜30 mL/（min·1.73 m²）. The patients with chronic kidney disease，40 patients with high risk of chronic kidney disease （with diabetes mellitus or hypertension） and 32 healthy subjects were compared with the testing of random urine microalbumin （U-mALb）/creatinine (Cr) ratio, random urine protein (UP)/Cr ratio, 24h-urine microalbumin（24h-UmALb），24h-urine protein（24hUP），serum urea，Cr，uric acid（UA） and cystatin C（CysC）. With the gold standard methods ［the cut-off value of 24h UP or eGFR was >150 mg or ＜60 mL/（min·1.73 m²），respectively］， the diagnostic sensitivity, specificity, negative predictive value, positive predictive value, negative and positive likelihood ratios and accuracy were calculated. Results　There were significant differences in the results of all tests（U-mALb/Cr, UP/Cr, 24hU-mALb, urea, UA and CysC） between the 3 groups of chronic kidney disease（P<0.05）. There were also significant differences in the results of all the tests except UA between the Group A of chronic kidney disease and the group with high risk of chronic kidney disease（P<0.05）. There only existed significant difference in the results of UA and CysC between the group of healthy subjects and the group with high risk of chronic kidney disease（P<0.05）. The diagnostic performances of U-mAlb/Cr, UP/Cr, 24 h U-mAlb, urea, UA and Cys C were as follows：negative likelihood ratios were 0.17，0.22，0.14，0.67，0.66 and 0.51；positive likelihood ratios were 7.73，78，28.7，34，3.73 and 5.00；accuracies were 0.87，0.88，0.92，0.66，0.65 and 0.74. Conclusions　The 24hU-mALb has the best accuracy in the diagnosis of chronic kidney disease. UP/Cr and U-mALb/Cr may be used as screening tests for the diagnosis of chronic kidney disease，because their accuracies of diagnostic performance for chronic kidney disease are closest to that of 24 h U-mALb, and eGFR may be mainly used in the classification of chronic kidney disease.

Application of enhanced cardiac troponin I determination in clinical diagnosis of acute coronary syndrome

ZHANG Guang-Hui;DENG Lin;FEI Ai-Hua;DAI Li-Hua;LU Yi-Wen;SHEN Li-Song

2012, 27(5):  345-348. 

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Objective　To observe the distribution of enhanced cardiac troponin I (cTnI)［high sensitive cTnI (hs-cTnI)］ in healthy subjects and patients with acute coronary syndrome (ACS), and investigate the application of hs-cTnI in the diagnosis of ACS.   Methods　A total of 80 ACS patients who had chest pain onset within 12 h, 26 stable angina (SA) patients and 60 healthy controls were enrolled. The hs-cTnI，conventional cTnI，blood routine tests, C reactive protein (CRP) and other indices were determined among the groups. The receiver operating characteristic (ROC) curve was used to analyze the diagnosis performance of hs-cTnI determination. Results　The concentrations of hs-cTnI had statistical significance among ACS, SA and control groups. The area under ROC curve (AUC) of hs-cTnI (0.986±0.014) was significantly higher than that of conventional cTnI (0.915±0.035, P=0.034). The AUC of hs-cTnI had no difference with those of hs-cTnI combined with CRP, neutropil granulocyte percentage/lymphocyte percentage ratio and white blood cell count (0.978±0.018,P>0.05). When the cut-off value was 0.02 ng/mL,the diagnosis performance was the best. Conclusions　Compared with the conventional cTnI, the hs-cTnI has diagnosis performance in early diagnosis of ACS. There is no diagnosis value in combined of inflammation indices.Objective　To observe the distribution of enhanced cardiac troponin I (cTnI)［high sensitive cTnI (hs-cTnI)］ in healthy subjects and patients with acute coronary syndrome (ACS), and investigate the application of hs-cTnI in the diagnosis of ACS.   Methods　A total of 80 ACS patients who had chest pain onset within 12 h, 26 stable angina (SA) patients and 60 healthy controls were enrolled. The hs-cTnI，conventional cTnI，blood routine tests, C reactive protein (CRP) and other indices were determined among the groups. The receiver operating characteristic (ROC) curve was used to analyze the diagnosis performance of hs-cTnI determination.  Results　The concentrations of hs-cTnI had statistical significance among ACS, SA and control groups. The area under ROC curve (AUC) of hs-cTnI (0.986±0.014) was significantly higher than that of conventional cTnI (0.915±0.035, P=0.034). The AUC of hs-cTnI had no difference with those of hs-cTnI combined with CRP, neutropil granulocyte percentage/lymphocyte percentage ratio and white blood cell count (0.978±0.018,P>0.05). When the cut-off value was 0.02 ng/mL,the diagnosis performance was the best.  Conclusions　Compared with the conventional cTnI, the hs-cTnI has diagnosis performance in early diagnosis of ACS. There is no diagnosis value in combined of inflammation indices.

The changes and clinical significance of serum visfatin and adiponectin levels in patients with coronary heart disease

JIAO Lu-Yang;WANG Yu;YUAN Yu

2012, 27(5):  349-351. 

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Objective　To investigate the relationship of serum visfatin， adiponectin and coronary heart disease with C reactive protein (CRP) and its possible mechanism.   Methods　A total of 270 patients with coronary heart disease confirmed by coronary angiography were enrolled. According to clinical manifestations, they were classified into 3 groups：stable angina pectoris （SAP） group (125 cases), unstable angina pectoris （UAP） group (109 cases) and acute myocardial infarction （AMI） group (36 cases). Another 20 healthy subjects were as control group. The serum visfatin and adiponectin levels were determined by enzyme-linked immunosorbent assay (ELISA). The levels of CRP, total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), fasting glucose (FPG), alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea nitrogen (BUN) and creatinine (Cr) were also determined. The multiple correlation analysis method was used to analyze the correlation of visfatin and adiponectin with other indices.  Results　In the coronary heart disease group, CRP and serum visfatin levels were significantly higher than those in control group（P<0.01）. The serum adiponectin level was lower than that of control group （P<0.01）. In AMI group， UAP group and SAP group, CRP， serum visfatin and adiponectin were statisically significant （P< 0.05）. The multiple correlation analysis showed that serum visfatin was positively correlated with CRP， TG and LDL-C （r=0.929，0.756 and 0.793, P<0.001）. The serum visfatin was negatively correlated with adiponectin and HDL-C （r=-0.351 and -0.699,P<0.001）. The serum adiponectin was negatively correlated with CRP, TG and LPL-C (r=-0.873，-0.639 and -0.799, P<0.001), and was positively correlated with HDL-C (r=0.681,P<0.001).  Conclusions　Serum visfatin and adiponectin may be risk factors for coronary heart disease， and they may interact with CRP，and be involved in the occurrence and development of coronary heart disease.

Value of serum cystatin C and creatinine in evaluating glomerular filtration rate in patients with chronic kidney disease ：a meta-analysis

MA Qing-Guang;LI Hui-Min;SHEN Li

2012, 27(5):  352-357. 

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Objective　To evaluate the diagnostic value of serum cystatin C (Cys C) and creatinine (Cr) in glomerular filtration rate (GFR) of chronic kidney disease patients. Methods　MEDLINE, EMbase, the Cochrane Library and other databases (1966 to 2010 September) were searched for collecting studies which evaluated the diagnostic value of serum Cys C and Cr in evaluating GFR in patients with chronic kidney disease. The heterogeneity was tested by the Cochrane Collaboration′s software RevMan 4.2.7. The summary receiver operating characteristic (SROC) curve and Meta-analysis were performed by Metadisc.  Results　A total of 231 relevant articles were searched, and 14 articles were included in the study. The study subjects were involved 2 169 patients. Meta-analysis showed that the cut-off value of GFR was 80-90mL/min, the Cys C heterogeneity was high ［P=0.098, I²=34.7%, summary sensitivity (SEN)=0.873, summary specificity (SPE)=0.791］, and Cr had no heterogeneity (P=0.56，I²=0.0%，SEN=0.740, SPE=0.867）. In 5 articles with various stages of GFR, Meta-analysis of GFR (60-90mL/min) group showed that Cys C and Cr had no heterogeneity {Cys C：P=0.708，I²=0.0%，odds ratio (OR)=7.04 ［95% confidence interval (CI): 4.54-10.92］, SEN=0.813，SPE=0.609，the area under the SROC curve (SAUC)=0.822 6；Cr：P=0.945，I²=0.0%，OR=9.4(95%CI:5.75-15.3), SEN=0.605，SPE=0.819，SAUC=0.826 3}. Conclusions　There are significant correlations between Cys C, Cr and GFR. As for early chronic kidney disease, CysC has more sensitivity but less specificity than Cr for evaluation of GFR. Further high quality researches on various stages of GFR in chronic kidney disease patients are still required.

Early changes of plasma N-terminal pro-B-type natriuretic peptide original in cardiac injury

ZHU Lanyun;MI Xiuhua;YANG Lianli;XU Laishun;CHE Xiaoqiong

2012, 27(5):  358-360. 

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Objective　To investigate the changes of plasma N-terminal pro-B-type natriuretic peptide (NT-proBNP) in hypertension and coronary heart disease and the cardiac function early injury in patients with diabetes mellitus and without abnormal renal function， and observe the change of NT -proBNP in hypertension and coronary heart disease before and after treatment. Methods　The 206 patients with hypertension and coronary heart disease and 65 patients with diabetes mellitus were classified into Ⅰ-Ⅱ grade group and Ⅲ-Ⅳ grade group according to the New York Heart Association (NYHA) heart pathological grading，and 37 patients without hypertension, coronary heart disease and diabetes mellitus in the department of bone traumatology were enrolled as controls. The NT-proBNP concentrations were detected. A total of 37 patients with hypertension and coronary heart disease were selected and compared for NT-proBNP concentrations before and after the treatment. The relationship of treatment and prognosis with NT-proBNP was observed. Results　The NT-proBNP levels in hypertension and coronary heart disease group and diabetes mellitus group were higher than those in the control group (P<0.01). The NT-proBNP levels in hypertension and coronary heart disease NYHA Ⅰ-Ⅱ grade group were higher than those in diabetes mellitus NYHA Ⅰ-Ⅱ grade group (P<0.05)，but there was no statistical difference in the NT-proBNP levels between hypertension and coronary heart disease NYHA Ⅲ-Ⅳ grade group and diabetes mellitus NYHA Ⅲ-Ⅳ grade group (P>0.05). The NT-proBNP levels in hypertension and coronary heart disease group after the treatment were significantly lower than those before the treatment (P<0.01). Conclusions　NT-proBNP can be used for the laboratory diagnosis of hypertension and coronary heart disease in cardiac function early injury, and associates with treatment and prognosis. NT-proBNP can reflect the change of cardiac function sensitively in the early development of diabetes mellitus.

Investigation on the complementation significance of combined cTnI，IMA and hs-CRP determination in coronary heart disease

WU Weihua;YI Jianhua

2012, 27(5):  361-363. 

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Objective　To investigate the application and clinical significance of combined determination of cardiac troponin I（cTnI），ischemia modified albumin（IMA） and high sensitive C reactive protein（hs-CRP） in coronary heart disease.   Methods　Enzyme-linked immunosorbent assay (ELISA) kits from the same company were used to measure the all markers. According to the measured results of cTnI were higher or not than the reference value (0.173ng/mL), 142 patients were classified into 2 groups ［cTnI non-increasing (<0.173 ng/mL) group: 91 cases, cTnI increasing (≥0.173 ng/mL) group: 51 cases］. A total of 78 healthy subjects were enrolled as controls. The combined determination of cTnI, IMA and hs-CRP was performed. Results　Among coronary heart disease patients in cTnI non-increasing group, their IMA and hs-CRP levels increased slightly compared with those in the control group（P<0.05）. The IMA and hs-CRP levels among coronary heart disease patients in cTnI increasing group increased significantly（P<0.01）. Among coronary heart disease paritents in cTnI non-increasing group, the IMA levels of 27 patients (30%) increased slightly, and the hs-CRP levels of 34 patients (37%) increased. Conclusions　The combined determination of cTnI，IMA and hs-CRP has complementation significance in diagnosis of myocardial ischemia and myocardial injury.

Flow cytometry application in analyzing T lymphocyte subsets of cerebrospinal fluid in patients with neural systemic diseases

CAI Bei;FENG Weihua;LI Lixin;WANG Lanlan

2012, 27(5):  364-370. 

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Objective　To analyze T lymphocyte and its subsets in cerebrospinal fluid (CSF) by flow cytometry (FCM), investigate the difference of T lymphocyte subset percentages in CSF of patients with different neural systemic diseases, and find the significant indicators in diagnosis and treatment monitoring. Methods　The T lymphocyte subsets in peripheral blood and CSF were determined in patients with neural systemic diseases. Results　Making plots based on side scaffer (SS) vs. CD45 was more accurate to define lymphocytes in CSF by FCM. FCM was only suitable for analyzing lymphocyte subsets in neural systemic immune diseases. The percentages of CD3⁺T lymphocytes, CD3⁺CD4⁺T lymphocytes and CD3⁺CD8⁺T lymphocytes in CSF of patients with inflammatory diseases of central nervous system or myelitis were the highest, those in patients with demyelinating diseases were the lowest. There was no significant difference in the percentage of T lymphocyte subsets of CSF among kinds of neural systemic diseases(P>0.05). The percentages of CD3⁺T lymphocytes and CD3⁺CD4⁺T lymphocytes in CSF from patients with bacterial meningitis were higher than those in patients with demyelinating diseases (P=0.032), and there was no statistical significance with those in patients with viral meningitis（P>0.05). The ratios of CD4/CD8 were significantly higher in CSF in all kinds of neural systemic diseases than those in peripheral blood. The ratio in CSF was the highest in patients with bacterial meningitis and myelitis, and was the lowest in demyelinating diseases with no statistical significance（P>0.05）. The correlation coefficient(r) was 0.324 between CSF IgG production index and the percentage of CD3 ⁺ CD4 ⁺ T lymphocytes in CSF (P=0.032). ConclusionsFCM for T lymphocyte subset analysis in inflammatory neural systemic diseases is more sensitive and helpful in analyzing the immune status of CSF than CSF routine examination. High percentage of CD3⁺CD4⁺T lymphocytes is closely related with neural systemic inflammatory diseases, but it is limited for FCM application in neurodegenerative diseases.

TSA induced cell cycle apoptosis in nasopharyngeal carcinoma CNE2 cells and its mechanisms

YANG Huimin;HUANG Lin;QIN Mengbin;ZHONG Yueyuan;LIU Xiaochun;YANG Xiaoli

2012, 27(5):  371-373. 

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Objective　To investigate the influence of trichostatin A (TSA) on cell apoptosis in nasopharyngeal carcinoma CNE2 cells and its mechanism.   Methods　CNE2 cells were incubated with TSA at different concentrations (300, 400 and 500 nmol/L). The effects of TSA on cell apoptosis were detected by flow cytometry. The expression of cyclin-dependent kinase inhibitor 1A (P21) was analyzed by reverse transcription-polymerase chain reaction (RT-PCR). Results　The apoptosis rates with different concentrations of TSA (300, 400 and 500 nmol/L) were 12.52%±1.57%，19.69%±3.00% and 19.63%±2.68% after 24 h treatment. The apoptosis rates were 24.50%±4.45%，36.24%±3.92% and 34.82%±6.45% after 48 h treatment. There were statistical significance compared with those of controls (0 nmol/L TSA, P<0.05). The expression level of p21 increased after TSA treatment (P<0.01). Conclusions　TSA markedly induces CNE2 cell apoptosis. Its mechanism would be related with the abnormal expression of p21 gene.

Detection and clinical significance of serum anti-nuclear antibodies in patients with hepatitis B virus infection

DONG Min;ZHOU Houqing;WU Jinbin;CHEN Biting

2012, 27(5):  374-375. 

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Objective　To detect the serum anti-nuclear antibodies (ANA) and investigate its clinical significance in patients with hepatitis B virus (HBV) infection.   Methods　A total of 120 patients with HBV infection and 82 healthy subjects were enrolled， and indirect immuno-fluorescent assay (IIF) was used to detect the serum ANA. Results　The positive rate of ANA in 120 patients with HBV infection (30.0%) was higher than that of healthy subjects (4.2%). ANA in patients with HBV infection was mainly depended on low titer(1∶100, 58.3%). The ANA granular fluorescence (41.7%) was highest in the patients with HBV infection. Conclusions　HBV infection can induce to produce a variety of low titer ANA， and may play a certain role in the diagnosis， differential diagnosis and treatment. The pathogenesis of HBV may have some relationship with autoimmune.

Clinical study on the level of plasma procalcitonin in patients with hepatic cirrhosis and ascites

DU Yiqiao;XU Gang;YANG Zhenhua

2012, 27(5):  376-378. 

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Objective　To observe the level of plasma procalcitonin (PCT) in patients with hepatic cirrhosis and spontaneous bacterial peritonitis (SBP).   Methods　A total of 30 patients with hepatic cirrhosis and ascites were enrolled， including 16 patients without SBP (the non-SBP group) and 14 patients with SBP (the SBP group), and 20 healthy subjects were enrolled as controls. The levels of plasma PCT and serum C reative protein (CRP) were measured by semi-quantitative immunochromatographic test. The levels at the first time determination were as baseline values. Results　The levels of plasma PCT and serum CRP significantly increased in SBP group 3h， 6h， 12h after SBP occured when compared with the baseline values (P<0.01)， and were significantly higher than those in the non-SBP group and controls (P<0.01). After treatment， the levels of plasma PCT in the survivals were (0.48±0.25) ng/mL in the SBP group and (0.49±0.14) ng/mL in the non-SBP group， and no statistical significance was found (P>0.05). They were significantly lower than the baseline values (P<0.01). The levels of plasma PCT in the dying patients kept on a high level after treatment， and that in the SBP group ［4 cases， (37.67±15.48) ng/mL］ was significantly higher than that in the non-SBP group ［4 cases，(14.75±7.89) ng/mL］. Conclusions　The level of plasma PCT is useful for the diagnosis of SBP in patients with hepatic cirrhosis and ascites. If keeping on a high level， it may indicate bacterial infection and a bad prognosis.

The significance of scatter diagram from Sysmex XE2100 hematology analyzer on abnormal lymphocyte detection

WANG Xiaoou;CHEN Xiaojian;SHU Kuangyi;FANG Peipei

2012, 27(5):  379-381. 

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Objective　To investigate the variation principle of DIFF scatter diagram of Sysmex XE2100 hematology analyzer on abnormal lymphocyte.   Methods　A total of 40 samples with the percentages of abnormal lymphocyte >10% and <10% having DIFF scatter diagram obvious changes were observed, and the variation principle of DIFF scatter diagram was summarized. Results　The samples with abnormal lymphocyte had abnormal scatter diagram by Sysmex-XE2100 hematology analyzer. The 13 samples which mainly existed monocytoid atypical lymphocyte showed abnormally homogeneous scatter diagram described as ellipsoidal or oblong above the scatter diagram of monocyte. Among 27 samples which mainly mixed with plasmacytoid atypical lymphocyte and blast-like atypical lymphocyte, there were 19 samples′scatter diagram turning into greyish white and extended upward in the area of lymphocyte and monocyte on the DIFF channel, and the other 8 samples′scatter diagram showed that lymphocyte, monocyte and neutrophile granulocyte in their respective area, but the scatter increased and overlapped around these areas. Conclusions　With the prompt indication of abnormal lymphocyte by the analyzer, the DIFF scatter diagram of hematology analyzer with microscopy on blood smear could reduce the chances of misdiagnosis and misidentified cases.

Investigation on the correlation of cerebrovascular disease thrombelastograph with fibrinogen

QIU Lijun;GU Qing;MENG Jun;SHEN Lisong

2012, 27(5):  382-385. 

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Objective　To investigate the relationship of thrombelastograph (TEG) parameters with fibrinogen（Fg） in patients with cerebrovascular disease.   Methods　A total of 470 patients with cerebrovascular disease［70 patients with cavity infarction and 400 patients with cerebral infarction（73 patients with pure cerebral infarction，200 cerebral infarction patients with high blood pressure，27 cerebral infarction patients with diabetes mellitus and 100 cerebral infarction patients with high blood pressure and diabetes mellitus）］ and 50 healthy controls were enrolled. TEG-5000 TEG analyzer （TEG-5000） was used to determine the parameters of K value，angle （A） value and MA value. The Fg level was detected by ACL-TOP automatic blood coagulation analyzer. The platelet （PLT） count and mean platelet volume（MPV）were detected by LH 750 automatic blood analyzer. Results　There was negative correlation between Fg level and K value（r＝－0.134，P=0.000）. There was positive correlation between Fg level and A value，MA value and PLT count（r＝0.210，P=0.000；r＝0.251，P=0.000；r＝0.122，P=0.000）. There was no correlation between Fg level and MPV（r＝－0.007; P=0.957）. The A value，MA value，PLT count and Fg level in cavity infarction group and cerebral infarction group were higher than those in the control group（P＜0.05，P<0.01）. MPV value in cerebral infarction group was higher than that in the control group（P＜0.05）, but there was no statistical significance between cavity infarction group and the control group (P>0.05). The K value had no statistical significance among cavity infarction group，cerebral infarction group and the control group (P>0.05). The MA value in cerebral infarction patients with diabetes mellitus was higher than that in patients with pure cerebral infarction and cerebral infarction patients with high blood pressure（P＜0.05）. The Fg levels in cerebral infarction patients with oliabetes mellitus and cerebral infarction patients with diabetes mellitus and high blood pressure were higher than those in cerebral infarction patients with high blood pressure（P＜0.05）. The other parameters had no difference between each group. Conclusions　In the process of cerebrovascular disease，there are changes of PLT quantity，quality，form and Fg level. Fg and TEG parameters have obvious correlation. TEG determination with the practical value can be one of the screening tests for cerebrovascular disease.

Significance evaluation of BV Blue method in diagnosis of bacterial vaginosis

JIANG Zhifeng;ZONG Weifang;SUN Peifang

2012, 27(5):  386-388. 

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Objective　To investigate the significance of BV Blue method in the diagnosis of bacterial vaginosis (BV), and provide rapid and reliable methods for diagnosing BV in clinic.   Methods　BV Blue method and Amsel method were used to determine 560 samples (<30 years old group, 31-40 years old group, 41-50 years old group and >50 years old group)， and the results were analyzed comparatively. Results　The positive rate of BV Blue method for diagnosing BV was 20.89%, and the positive rate of Amsel method was 24.82%. There was no significant difference （P>0.05）. The sensitivity， specificity and accuracy of BV Blue method for diagnosing BV was 97.86%， 77.79% and 92.85%, respectively. BV positive rate increased with age， and the highest rate was 28% when the age was >50 years old. Conclusions　BV Blue method is simple and rapid, and has high sensitivity and accuracy. It suitable for the rapid diagnosis of BV.

Relationship investigation on idiopathic thrombocytopenic purpura with cytomegalovirus infection

CHEN Hu;XIAO Jianhua.

2012, 27(5):  389-392. 

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Objective　To study the relationship between the idiopathic thrombocytopenic purpura（ITP）and the infection of cytomegalovirus（CMV）, the clinical symptom of ITP，and the curative effect of ITP patients with CMV infection.   Methods　The 153 cases of ITP in-patients in hospital were studied. All patients were measured the antigens of CMV PP65 of white blood cell in peripheral blood. The general data, clinical manifestation, laboratory examination and treatments of 3 investigated groups were analyzed by cytomorphology and statistics methods. Results　The infective rate of CMV in ITP patients was 20.3%.The average age of ITP patients with CMV infection was 20.03±11.7. The ITP patients with CMV infection were inclined to suffer from skin hemorrhage. In the ITP patients with CMV infection, the average of marrow megacaryocyte numbers was lower than that of the negative controls，and megakaryocytic maturation lag was earlier. The general effective ratio in ITP patients with CMV infection treated with glucocorticoids was 35.5%. The total effective ratio of combined therapy，as glucocorticoids and ganciclovir (GCV), in ITP patients with CMV infection was 85.0%. Conclusions　CMV may be one of the viruses causing ITP disease. The ITP patients with CMV infection are inclined to occur skin hemorrhage. The marrow megacaryocyte productive reaction of ITP patients with CMV infection was inferior to that of negative controls，and megakaryocytic maturation lag is earlier. The curative effect of glucocorticoids is low. The combined therapy of glucocorticoids and GCV could improve curative effect.

The detective value of high-risk human papillomavirus genotype 16 and 18 DNA in screening cervical lesions

ZHOU Bin;ZUO Xinhua;FU Xinwen

2012, 27(5):  393-395. 

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Objective　To evaluate the detective value of high-risk human papillomavirus (HPV) genotype 16 and 18 (HPV16,18) in screening cervical lesions.   Methods　By histological biopsy, 209 cases were cancer and percancerous lesions ［cervical intraepithelial neoplasia (CIN) Ⅰ, CIN Ⅱ, CIN Ⅲ and invasive cervical cancer］, and 296 cases were inflammation, polyp and other benign lesions. HPV16，18 DNA and liquid-based cytology were used to detect the cases with cancer and percancerous lesions by histological biopsy. Comparing the results of the 2 methods， HPV16，18 infectious rates， sensitivity， specificity and negative predictive value were statistically analyzed. Results　Of 209 patients with cancer and precancerous lesions，158 cases were positive of HPV16，18，and 128 cases were positive of liquid-based cytology ［low-grade squamous intraepithelial lesion (LSIL), atypical squamous cells, cannot exclude a high-grade squamous intraepithelial lesion (ASC-H), high-grade squamous intraepithelial lesion (HSIL) and squamous cell carcinoma］. Among the 296 patients with benign lesions, 43 cases were positive of HPV16,18, and 2 cases were positive of liquid-based cytology. The results of cancer and precancerous lesions showed that the positive rate of HPV16，18 DNA was higher than that of liquid-based cytology (χ²＝12.69，P<0.01). The sensitivity， specificity and negative predictive value of HPV16，18 DNA were 75.6%， 85.5% and 83.2% respectively, and 61.2%, 99.3% and 78.4% of liquid-based cytology. The HPV16,18 positive rate of CIN Ⅱ was obviously higher than that of CINⅠ(χ²＝6.69，P<0.05). Conclusions　The occurrence of cervical cancer and percancerous lesions is closely related to HPV16,18 infection. Compared with liquid-based cytology，HPV16，18 DNA in cervical cancer and percancerous lesions has a higher sensitivity，lower specificity and higher negative predictive value. HPV16，18 DNA has an important clinical value for screening cervical benign lesions, and it is better than liquid-based cytology.

The characteristic of nosocomial infection pathogens and their drug-resistance analysis

FAN Qianyan;SHEN Min;TANG Yuanyuan;TAN Qiuwen

2012, 27(5):  396-399. 

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Objective　To investigate the distribution and drug-resistance rate of pathogens in nosocomial infection, in order to provide scientific reference for the reasonable clinical application of antibiotics.   Methods　The bacterium identifications and drug sensitivity assay in various kinds of clinical samples (47 469 cases) were performed by Microscan Walkaway 40SI system and in vitro drug sensitivity assay. Results　A total of 7 445 pathogens had been isolated form the 47 469 samples. The positive rate of Gram-negative bacteria was 81.87%， and these bacteria were mainly composed of Escherichia coli (23.36%)，Klebsiella pneumonia (16.70%)，Pseudomonas aeruginosa (8.77%) and Acinetobacter baumannii (7.64%). The positive rate of Gram-positive bacteria was 18.13%， and these bacteria were mainly composed of Staphylococcus aureus (8.62%)，Enterococcus faecalis (3.99%) and Enterococcus faecium (2.15%). Pathogens causing nosocomial infections had strong drug-resistance and were varied in drug-resistance spectrum. Conclusions　Timely information about pathogen distribution and trend of drug-resistance variation can guide the reasonable application of antibiotics in clinical practice，and eliminate or largely reduce the unreasonable use or misuse of antibiotics， therefore reducing the nosocomial infections.

Development and evaluation of Real-time PCR and TaqMan-MGB Probe assay for detecting of apolipoprotein A5 gene

CHEN Juan;LI Lei;WEI Hongxia;ZHANG Kui

2012, 27(5):  400-403. 

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Objective　To establish fluorescence quantitation polymerase chain reaction (PCR) based on TaqMan-MGB probe for detecting apolipoprotein A5（apo A5）gene. Methods　The assay, which was based on specific primers and TaqMan-MGB probe from apo A5 gene, was performed to reconstruct DNA fragment of pPCR-Script-apoA5. The fluorescence quantitation PCR with TaqMan-MGB probe and the standard curve were established. The sensitivity， specificity and repeatability of the TaqMan-MGB probe fluorescence quantitation PCR were determined. Results　A fine linear relationship between the threshold cycle (Ct) values of the developed standard curve and the copy number of template was observed (r=0.998). The sensitivity of the assay was 10³ copies/μL, the amplification efficiency of assay was 110.2%, and the repeatability was good.Conclusions　The fluorescence quantitation PCR based on TaqMan-MGB probe for detecting the apo A5 gene is successfully developed with high sensitivity，specificity and repeatability.

Research on vWF gene polymorphisms in Han population patients with acute cerebral infarction Disease in Kunming

OUYANG Hongmei;ZHANG Qin;ZHAI Ming;JIANG Yaxian;ZHANG Lei;NIU Hua

2012, 27(5):  404-407. 

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Objective　To clarify the distribution of the von Willebrand factor (vWF) gene intron 19 MspI polymorphism among Han population in Kunming，and investigate the relationship of vWF gene intron 19 MspI polymorphism with acute ischemic cerebral infarction(AICI). Methods　A total of 119 patients with AICI and 117 healthy controls were enrolled. Polymerase chain reaction-restriction fragment length polymorphism（PCR-RFLP） was used to determine the vWF gene intron 19 MspI polymorphism. Results　Among the Han population in Kunming, the vWF gene intron 19 MspI M+/M+，M+/M-and M-/M-genotype frequencies were 0.034，0.470 and 0.496. M+ and M- allele frequencies were 0.269 and 0.731. M+/M+，M + / M-and M-/ M- genotype frequencies in AICI group were 0.059, 0.286 and 0.655，and M + and M- allele frequencies were 0.202 and 0.798, respectively. M+/M- genotype frequencies in AICI group were lower than those in control group, and M-/M- genotype frequencies were higher than those in control group. The frequencies between AICI and control groups had significant difference (P<0.05). Conclusions　The vWF gene intron 19 MspⅠ restriction site genotype distributions among Han population in Kunming are quite different comparing with Gaucasian populations，and the other ethnic populations in Kunming has some differences. Acute cerebral infarction may be associated with vWF gene intron 19 MspⅠ polymorphisms.

The construction and confirmation of HIV-1 electrochemically active–inactive switching molecular beacon

LI Bo;GU Dayong;SITU Bo;LI Zhengliang;YAN Xiaohui;LIU Qinlan;ZHENG Lei;WANG Qian

2012, 27(5):  408-411. 

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Objective　To construct human immunodeficiency virus-1 (HIV-1) electrochemically active-inactive switching molecular beacon(CAs-MB) and confirm its structure and function.   Methods　CAs-MB targeted conserved region of HIV-1 gag gene was prepared and purified. The structure was confirmed by Fourier transform-infrared spectroscopy (FT-IR) and cyclic voltammetry (CV), while its function of recognizing targets was evaluated by differential pulse voltammetry (DPV). Results　The variations of FT-IR spectra of free carminicacid (CA) and CAs-MB indicated that CA had covalently linked onto the ends of MB. The results of CV proved that both ON and OFF statuses of CAs-MB were existing. DPV results illustrated that CAs-MB was effective in signaling the presence of complementary target and discriminating targets in electrochemical gene biosensor that differ by a single nucleotide. Conclusions　CAs-MB opens a new detective technique and can be expanded to electrochemical gene biosensor to perform HIV-1 gag gene specificity detection.

Research on the correlation of PCSK9 gene I474V polymorphism with coronary artery disease

LIN Yongping;MENG Yanhui;ZHENG Weidong;LIU Zhongmin

2012, 27(5):  412-415. 

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Objective　To investigate the distribution characteristics of the I474V loci of proprotein convertase subtilisin lkexin type 9 (PCSK9) gene in coronary artery disease (CAD) patients and healthy subjects, study the correlation of blood lipid levels， the development of CAD and the severity of coronary lesions，and provide the reference to elucidate the pathogenesis and genetic risk assessment of CAD.   Methods　According to the results of coronary angiography, blood samples from 288 CAD patients and 300 healthy controls were collected. I474V polymorphism loci in PCSK9 gene was detected by bi-directional polymerase chain reaction (PCR) amplification of specific alleles (Bi-PASA PCR). The results were further confirmed by cycle sequencing. Serum levels of total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) were measured by enzymatic methods. Results　Ⅱ and Ⅳ genotypes were dominant in the I474V polymorphism loci of PCSK9 gene. No significant difference was found in allele frequencies of I474V loci between CAD patients and healthy controls (P>0.05). There was statistical significance in TC, LDL-C and HDL-C levels between CAD patients and healthy controls (P<0.05), and the TG level had no statistical significance (P>0.05). Serum levels of TC and TG (P<0.05), but not HDL-C and LDL-C (P>0.05) were statistically different between Ⅱ and Ⅳ genotypes in CAD patients. Subjects with I474V polymorphism of PCSK9 gene had significantly higher serum levels of TC and TG than subjects without I474V polymorphism. The number of coronary lesions was more in patients with Ⅳ genotype than Ⅱ genotype with statistical difference (P<0.05). Conclusions　Ⅱ and Ⅳ genotypes are dominant in the I474V polymorphism loci of PCSK9 gene. The genotype of PCSK9 gene I474V polymorphism is correlated with serum increasing levels of TC and TG and the severity of coronary lesions in CAD patients.

Analysis of pathogen causing hand-foot-mouth disease by real-time fluorescence PCR

ZHANG Qun;YANG Lei;CAO Weifeng

2012, 27(5):  416-418. 

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Objective　To detect the pathogens by real-time fluorescence quantitation polymerase chain reaction (FQ-PCR) and provide the reference for the prevention of hand-foot-mouth disease. Methods　Throat swab and vesicle fluid were collected from 189 suspected children with hand-foot-mouth disease and 39 children with close contact to hand-foot-mouth disease patients in Anhui county and its neighborhood area. Enterovirus (EV), enterovirus 71 (EV71) and coxsakievirus A16 (CoxA16) were detected by FQ-PCR, and the results were compared with the results of enzyme-linked immunosorbent assay (ELISA). Results　By FQ-PCR, 141 children among the 189 suspected children with hand-foot-mouth disease were detected EV (positive rate: 74.6%), including 91 children between 0 and 3 years old and 50 children between 3 and 5 years old. EV detection rate of 3-5 years old group was obviously lower than that of 0-3 years old group (P<0.05). There were 135 EV71 positive samples and 65 CoxA16 positive samples. The positive rate of throat swab samples was 74.5% (120/161), and that of vesicle fluid samples was 82.1% (23/28). There were 76(40.2%) EV positive samples by ELISA. The positive rate of EV in 39 children with close contact to hand-foot-mouth disease patients by FQ-PCR was 33.3%(13/39). The positive rate by ELISA was 17.9%(7/39). Conclusions　FQ-PCR has high specificity, sensitivity, efficiency and speed, which contributes to its important value as detection method of pathogen causing hand-foot-mouth disease and epidemiological study.