Table of Content

20 January 2014, Volume 29 Issue 1

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Pay closer attention to the clinical research for cellular immunity of subjects infected with hepatitis B virus

ZHU Qinghui

2014, 29(1):  1-3.  DOI: 10.3969/j.issn.1673-8640.2014.01.001

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The cellular immunity of subjects infected with hepatitis B virus(HBV) is the critical factor for their recognizing and clearing virus (acute self limiting infection) or showing immune tolerance (chronic persistent infection), and is tightly correlated with the disease prognosis and outcome. Thus, the clinical monitoring cellular immunity of subjects infected with HBV is very necessary. The characteristics of the review and papers published in the special subject, Clinical Monitoring for the Cellular Immunity of Subjects Infected with Hepatitis B Virus, are evaluated. The development of studies in basic immunology will provide wider field for studies in clinical immunology. The closer attention of physicians and experimenters to the clinical research for cellular immunity of subjects infected with HBV is being expected.

The roles of T lymphocytes and natural killer cells in chronic infection of hepatitis B virus

LIANG Yan, WANG Hao, ZHONG Renqian.

2014, 29(1):  4-9.  DOI: 10.3969/j.issn.1673-8640.2014.01.002

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Chronic infection caused by hepatitis B virus (HBV) is a serious infectious disease to threaten health in our country. The immunological mechanism of the chronicity has been the focus. The paper summarizes the development of the roles of T lymphocytes and natural killer(NK) cells in chronic infection of HBV, in order to help with developing insights into the cellular immune in chronic infection of HBV and to provide a new thought to diagnose and treat the chronic infection of HBV.

Primary analysis on the function of peripheral blood HBV-specific CD8⁺T cells from patients with chronic hepatitis B

CAI Gang, WU Beiying, FAN Zhenjia, SHEN Qian.

2014, 29(1):  10-14.  DOI: 10.3969/j.issn.1673-8640.2014.01.003

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Objective To evaluate the function of peripheral blood hepatitis B virus(HBV)-specific CD8⁺ T cells from patients with chronic hepatitis B(CHB), and assess its relationship with clinical relative parameters. Methods The HBV-specific CD8⁺ T cells were detected by staining of major histocompatibility complex (MHC)-peptide tetramers in patients with CHB. The HBV-specific CD8⁺ T cells were analyzed for cytokine secretion ability. The relationship of HBV-specific CD8⁺ T cells with clinical biochemical and viral indices was analyzed. Results A total of 9 of 35 patients with human leukocyte antigen (HLA)-A02 genotype or A24 genotype had HBV-specific CD8⁺ T cells recognized by tetramers in peripheral blood. The HBV-specific CD8⁺ T cells expressed significantly lower interleukin 2(IL-2) and interferon gamma(IFN-γ) than their counterpart CD8⁺ T cells. The patients who can be found HBV-specific CD8⁺ T cells had higher level of alanine aminotransferase(ALT) and lower HBV DNA load than those without HBV-specific CD8⁺ T cells. Results Persistent stimulation of HBV antigen is one of the causes of HBV-specific CD8⁺ T cell dysfunction in patients with CHB. The patients who can be found HBV-specific CD8⁺ T cells in peripheral blood could have strong anti-virus immunity.

The proportion of Th17 cells and IL-17 expression levels in peripheral blood of patients with chronic hepatitis B and their clinical significance

DING Qingli, TANG Gusheng, HE Zhengwen, SHEN Qian, QIN Yanghua.

2014, 29(1):  15-20.  DOI: 10.3969/j.issn.1673-8640.2014.01.004

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Objective To analyze T helper 17(Th17) cell proportion and interleukin 17(IL-17) levels in peripheral blood of patients with chronic hepatitis B(CHB) and healthy controls, and to evaluate their correlations with disease severity, serum hepatitis B virus(HBV) DNA load and liver injury markers, in order to investigate Th17 cells′ role in the pathogenesis of CHB. Methods The Th17 cell proportion and IL-17 levels in peripheral blood of 93 CHB patients (the mild, moderate and severe CHB patients had 21, 37 and 35 cases, respectively) and 28 healthy controls were determined by flow cytometry and enzyme-linked immunosorbent assay(ELISA). The mRNA expression levels of IL-17 and RAR-related orphan nuclear receptor C (RORC) of purified CD4⁺ T cells after stimulating with anti-CD3 and anti-CD28 antibodies were analyzed by real-time fluorescence polymerase chain reaction(PCR). The IL-17 levels in the supernatant of purified CD4⁺ T cells after stimulating with anti-CD3 and anti-CD28 antibodies were evaluated by ELISA. The data were analyzed statistically, and their relations with serum HBV DNA load, liver injury markers, and so on were analyzed. Results Compared with those of healthy control group, the proportion of Th17 cells in peripheral blood of CHB group was significantly higher, and increased with disease severity. The Th17 cell proportion, IL-17 levels and RORC mRNA levels in severe CHB group were significantly higher than those in moderate and mild CHB groups and healthy control group (P<0.05). There was no statistical significance in serum IL-17 levels among the groups(P>0.05). The IL-17 levels in the supernatant of purified CD4⁺ T cells after stimulating with anti-CD3 and anti-CD28 antibodies from CHB group were significantly higher than those from healthy control group(P<0.05), and the IL-17 levels in the supernatant from severe CHB group were higher than those from moderate and mild groups(P<0.05). The Th17 cell proportion in peripheral blood of CHB group showed a positive correlation with serum alanine aminotransferase(ALT) level (P<0.01), and showed no correlation with serum HBV DNA load. Results The increased Th17 cell proportion in CHB patients may be the one of the factors for liver inflammatory response and liver injury and cause the progression of CHB.

The detections of peripheral blood Th17 cells and Treg cells in HBV infected patients and their significance

HU Yueming, MA Li, LI Hong.

2014, 29(1):  21-25.  DOI: 10.3969/j.issn.1673-8640.2014.01.005

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Objective To investigate the percentage changes of T helper 17 (Th17) cells and regulatory T (Treg) cells in the peripheral blood of hepatitis B virus(HBV) infected patients and their correlations with liver injury after HBV infection. Methods The peripheral blood of 34 asymptomatic HBV carrier(ASC), 50 chronic hepatitis B(CHB) patients and 32 hepatocellular carcinoma(HCC) patients were collected. A total of 30 healthy controls(control group) were enrolled. The percentages of Th17 and Treg cells were detected by flow cytometry. Alanine aminotransferase (ALT) and aspartate aminotransferase(AST) were detected by Bayer ADVIA 1650 automatic biochemistry analyzer. Results Compared with control group (1.13%±0.37%), the percentage of Th17 cells in the peripheral blood of ASC group (1.30%±0.28%) was higher with no statistical significance (P> 0.05), but it obviously increased in CHB group (2.06%±1.15%) and HCC group (5.89%±3.31%, P<0.01). The percentage of Th17 cells in HCC group was higher than that in CHB group (P<0.05). The percentage of Th17 cells was positively correlated with ALT and AST [coefficients of correlation(r) were 0.682 and 0.637, P<0.01]. Compared with control group (7.18%±2.28%), the percentage of CD4⁺CD25⁺CD127^lowTreg cells in the peripheral blood of ASC group (7.72%±2.05%) was higher with statistical significance (P> 0.05), but it increased obviously in CHB group (9.01%±2.92%) and HCC group (10.86%±1.77%, P<0.01), and that in HCC group was higher than that in CHB group(P<0.05). The correlation of Treg cells was positive with ALT and AST (r=0.452, P<0.01; r=0.307, P<0.05). The correlation of Th17 cell percentage was positive with Treg cell percentage (r=0.327, P<0.01). The rate of Th17/Treg in control group was the lowest (0.17±0.16), that in ASC group(0.18±0.13) increased slightly, and there was no statistical significance between control group and ASC group (P> 0.05). However, there was obviously increasing in CHB group (0.23±0.19, P<0.05) and HCC group (0.52±0.54, P<0.01). Results In HBV infected patients, the percentages of Th17 and Treg cells increase. As for the immune balance of Th17 and Treg cells is broken, the increasing of Th17 cells mediates immune injury and induces the inflammation of liver cells.

Research on the relationship between serum cytokine levels and prognosis in patients with HBV-related acute-on-chronic liver failure

LUO Guangcheng, HUANG Yunli, YAN Huiping, FENG Xia, LIN Fang, FAN Quming, ZHANG Guoyuan.

2014, 29(1):  26-30.  DOI: 10.3969/j.issn.1673-8640.2014.01.006

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Objective To observe the changes of serum cytokine levels in patients with hepatitis B virus (HBV)-related acute-on-chronic liver failure(ACLF), and investigate the role of cytokines in the prognosis of HBV-related ACLF. Methods A total of 24 HBV-related ACLF patients(13 cured cases and 11 dead cases), 30 patients with chronic hepatitis B(CHB) and 15 healthy controls were enrolled. The serum levels of interleukin(IL)-2, IL-4, IL-6, IL-8, IL-10, granulocyte-macrophage colony stimulating factor(GM-CSF), gamma interferon (IFN-γ) and tumor necrosis factor alpha(TNF-α) were detected by Luminex liquid chip technology, and the relative Results were analyzed. Results The levels of IL-6, IL-8 and TNF-α in ACLF group were higher than those in CHB group and healthy control group (P<0.01), and those in CHB group were higher than those in healthy control group (P<0.05). IL-6, TNF-α and model for end stage liver disease (MELD) score in ACLF dead group were significantly higher than those in ACLF cured group(P<0.05). The levels of TNF-α and IL-6 were negatively correlated with prothrombin activity(PTA)(r=－0.712, P<0.001; r=－0.521, P=0.009). Furthermore, the levels of TNF-α and IL-6 were positively correlated with MELD score(r=0.491, P<0.015; r=0.379, P=0.048). There was no correlation among cytokines, HBV DNA and hepatitis B surface antigen(HBsAg). Results In HBV-related ACLF patients, various serum cytokines increase significantly. Furthermore, the levels of IL-6 and TNF-α increase with the severity of disease, and they are helpful in disease progression and prognosis.

Reference ranges of reticulocyte parameters of adults in Shanghai

JIN Shuo, DAI Min, ZHANG Jun, YANG Yan, SHEN Wei, YING Chunmei, FAN Zhuping.

2014, 29(1):  31-33.  DOI: 10.3969/j.issn.1673-8640.2014.01.007

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Objective To investigate the reference ranges of reticulocyte absolute value(RET#) and percentage(RET%) and its grouping parameters of adults in Shanghai. Methods By XE-2100 automatic hematology analyzer, the reticulocyte and it relative parameters were determined among 2 000 healthy subjects. There were 1 116 males and 884 females. A total of 2mL venous whole blood samples were collected for anticoagulation by ethylene diamine tetraacetic acid-dipotassium (EDTA-K₂). The samples were determined within 2h after collection. Statistical analysis was performed by SPSS11.0 software. Results The reference ranges of RET% were (0.61-1.87)% in males and (0.60-1.81)% in females, respectively. The RET# were (0.030-0.096)×10¹²/L in males and (0.025-0.082)×10¹²/L in females. The retios for low fluorescent reticulocyte(LFR), middle fluorescent reticulocyte(MFR) and high fluorescent reticulocyte(HFR) were (80.8-97.1)%, (2.8-18.1)% and (0.0-2.3)% in males and(82.3-97.9)%, (2.0-15.9)% and (0.0-1.8)% in females, respectively. By classifying the healthy subjects as age, there was no statistical significance in RET# and RET% (P=0.138 and P=0.061). However, there was statistical significance for grouping parameters (P<0.01). Results The reference ranges of reticulocyte parameters are established for adults in Shanghai. The clinical application significance of reticulocyte parameter determination is improved.

Relationship of coagulation factor Ⅴ and Ⅷ and tissue factor with type 2 diabetes mellitus combined with coronary heart disease

WANG Xiao, ZHU Junquan.

2014, 29(1):  34-37.  DOI: 10.3969/j.issn.1673-8640.2014.01.008

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Objective To investigate the application significance of coagulation factor Ⅴ and Ⅷ activities (FⅤ:C and Ⅷ:C) and tissue factor(TF) in diagnosis of type 2 diabetes mellitus (T2DM) combined with coronary heart disease(CHD). Methods FⅤ:C, FⅧ:C, TF, von Willibrand factor(VWF) activity and platelet aggregation rate(PAR) were measured in 85 patients with T2DM combined with CHD [T2DM with CHD group, including T2DM with acute myocardial infarction (AMI) group (19 cases), T2DM with old myocardial infarction (OMI) group (23 cases), T2DM with unstable angina pectoris (UAP) group (25 cases) and T2DM with stable angina pectoris(SAP) group (18 cases)], 29 T2DM patients without complication (single T2DM group) and 24 healthy controls (control group). The correlation between FⅤ:C, FⅧ:C and TF with VWF and PAR was analyzed. Results The levels of FⅤ:C, FⅧ:C and TF in T2DM with CHD group and single T2DM group were remarkably higher than those in control group (P<0.001), and those in T2DM with CHD group were higher than those in single T2DM group(P<0.01). In T2DM with AMI group and T2DM with UAP group, the levels of FⅤ:C, FⅧ:C and TF were markedly higher than those in single T2DM group(P<0.001), and those in T2DM with AMI group were higher than those in T2DM with UAP group(P<0.01). In T2DM with OMI group and T2DM with SAP group, the TF levels were significantly higher than those in single T2DM group(P<0.01), but there was no statistical significance among the 3 groups for FⅤ:C and FⅧ:C levels(P>0.05).There was a significantly positive correlation of FⅤ:C, FⅧ:C and TF levels with VWF and PAR levels(P<0.01). Results FⅤ:C, FⅧ:C and TF can be as the indicators for T2DM with CHD. TF is useful for the evaluation of disease development in patients with T2DM with CHD.


Analysis on the drug resistance of Enterococcus faecalis and Enterococcus faecium isolated from the midstream urine of patients with urinary tract infection in Antu Hospital

BAO Caili, FAN Qianyan, WU Di, TANG Yuanyuan, TAN Qiuwen, ZHANG Xuelian.

2014, 29(1):  38-41.  DOI: 10.3969/j.issn.1673-8640.2014.01.009

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Objective To analyze the drug resistance and gene of Enterococcus faecalis and Enterococcus faecium isolated from the midstream urine of patients with urinary tract infection in Antu Hospital, and to provide the reference for clinical medication. Methods Enterococcus faecalis (23 isolates) and Enterococcus faecium (18 isolates) were isolated and collected from the midstream urine of patients with urinary tract infection. The bacteria were identified by API, and drug sensitivity test was done by Kirby-Bauer disk diffusion method. Drug resistance genes were also detected by polymerase chain reaction(PCR). Results The positive rates of tetM, ermB, aac(6′)/aph(2′), ant(6)-Ⅰ and aph(3′)-Ⅲ genes were 65.2%, 82.6%, 100.0%, 100.0% and 100.0% in the 23 isolates of Enterococcus faecalis. In the 18 isolates of Enterococcus faecium, the positive rates of tetM, ermB, aac(6′)/aph(2′), ant(6)-Ⅰ and aph(3′)-Ⅲ genes were 55.5%, 55.5%, 100.0%, 94.4% and 100.0%. The drug resistance rates of Enterococcus faecalis to panicillin, tetracyline, ciprofloxacin, levofloxacin and erythomycin were >50.0%. The drug resistance rates of Enterococcus faecium to penicillin, ampicillin, ciprofloxacin, levofloxacin and erythomycin were 100.0%. Results The positive rate of drug resistance genes is high in Enterococcus isolated from clinic, which has a high multi-drug resistance.

Analysis on the NA gene variation of influenza A (H1N1) virus from Guangzhou in 2010

XU Shasha, CHANG Yanmin, XU Lin, FENG Fashen, HE Xia, WANG Zhu, ZHANG Dingmei, CAO Kaiyuan

2014, 29(1):  42-48.  DOI: 10.3969/j.issn.1673-8640.2014.01.010

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Objective To compare the variations of neuraminidase (NA) gene of influenza A (H1N1) virus from Guangzhou in 2010 and NA gene of influenza A(H1N1) virus from Chinese mainland in 2009, and to provide the reference for the surveillance and prevention of influenza A (H1N1) virus. Methods Specimens were collected from patients with febrile respiratory tract symptoms from Guangzhou in 2010. The specimens were determined with influenza A (H1N1) virus specificity primers by polymerase chain reaction (PCR). The fragment of NA gene was amplified. After sequencing, the Results were compared with influenza A (H1N1) virus in 2009.The drug resistance sites and glycosylation sites were analyzed by biometric software. Results A total of 1 194 specimens were collected, and 327 influenza A virus strains were detected, including 6 strains were identified with influenza A (H1N1) virus. Compared with influenza A (H1N1) virus in 2009, there were 16 amino acid sites with variation. There were 3 amino acid sites relating with NA activity. However, the amino acid site 222 located in the NA activity sites. Results NA gene of the 6 strains of influenza A (H1N1) virus are amplified and sequenced successfully, and H275Y drug resistant site is not found. The amino acid site 222, 228 and 425 locating in NA activity sites are found with mutations in 3 strains. Therefore, the monitoring should be strengthened.

Investigation on the gene frequency of Diego and Dombrock blood group systems of Han population in Jiangxi province

XIAO Li, YOU Rong

2014, 29(1):  50-52.  DOI: 10.3969/j.issn.1673-8640.2014.01.012

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Objective To investigate the rare Diego(Di) and Dombrock(Do) blood group systems of 200 random blood donors of Han nationality in Jiangxi province, and to obtain the appropriate gene frequency data for clinical blood transfusion. Methods A total of 200 random blood donors of Han nationality in Jiangxi province were enrolled, and polymerase chain reaction-sequence specific primer (PCR-SSP) was used to detect Di and Do blood group systems. Results In the 200 cases, there were 2 types of gene phenotypes of Di blood group, Di^a-b+(196 cases) and Di^a+b+(4 cases). Di^a+b-and Di^a-b- were not detected. The gene frequency of Di^a was 0.01, and the gene frequency of Di^b was 0.99.Do blood group identified 3 gene phenotypes, Do^a+b- (2 cases), Do^a+b+ (16 cases) and Do^a-b+ (182 cases). The gene frequency of Do^a was 0.05, and the gene frequency of Do^b was 0.95. Results The gene phenotypes of Di blood group were Di^a-b+ mainly and Di^a+b+, and Di^a+b- and Di^a-b- were not detected. The gene phenotypes of Do blood group were Do^a-b+ mainly, Do^a+b+ and Do^a+b-.

Relationship of seminal plasma neutral alpha-1, 4-glycosidase activity with seminal main parameters and seminal hyaluronidase activity in infertile patients

MA Xiaoping, GAO Xiaoqin, YANG Yanping, YANG Zhe.

2014, 29(1):  53-56.  DOI: 10.3969/j.issn.1673-8640.2014.01.013

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Objective To investigate the relationship of seminal plasma neutral alpha-1, 4- glycosidase (α-1, 4-G)activity with seminal main parameters and seminal hyaluronidase(HYD) activity(HYD positive rate and HYD activity intensity) in infertile patients. Methods A total of 52 infertile patients′ semen, who were between 24 and 40 years old, were collected, and 12 healthy controls were enrolled. The semen quality of each semen specimen was analyzed by sperm quality analysis instrument and computer-assisted sperm analysis. The enzyme method was used to detect the neutral α-1, 4-G activity. The HYD positive rate and HYD activity intensity were determined by improved fixed-substrate film method. According to the Results of neutral α-1, 4-G activity, the 52 infertile patients were classified into 39 cases of neutral α-1, 4-G activity normal group (activity normal group) and 13 cases of neutral α-1, 4-G activity abnormal group (activity abnormal group). Results There were significantly positive correlations of neutral α-1, 4-G activity with sperm concentration, sperm viability and HYD activity(HYD positive rate and HYD activity intensity) (r=0.816, 0.890, 0.872 and 0.847, P< 0.01). There was negative correlation with sperm malformation rate (r=-0.579, P<0.01). Sperm concentration, sperm viability and HYD activity(HYD positive rate and HYD activity intensity in infertile group were lower than those in control group(P<0.01). The sperm malformation rate in activity abnormal group was higher than that in activity normal group(P<0.01). Results The seminal plasma neutral α-1, 4-G activity could affect sperm concentration, sperm viability, sperm malformation rate and HYD activity (HYD positive rate and HYD activity intensity).

Investigation on the clinical significance of BNP and cTnI detections in AMI patients with non-diabetic irritable hyperglycosemia

CHEN Donglian, ZENG Fanpeng, DU Zhilin, LUO Weiping, LIANG Miaozhi, YANG Ying.

2014, 29(1):  57-60.  DOI: 10.3969/j.issn.1673-8640.2014.01.014

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Objective To study the roles of B-type natriuretic peptide (BNP) and cardiac troponin I (cTnI) detections in the recent prognosis evaluation of acute myocardial infarction (AMI) patients with non-diabetic irritable hyperglycosemia. Methods According to the Results of fasting blood glucose test, 122 AMI patients were enrolled and classified into 2 groups, AMI patients with non-diabetic irritable hyperglycosemia (experimental group, 68 cases) and AMI patients without non-diabetic irritable hyperglycosemia (control group, 54 cases). The BNP and cTnI levels were detected after the onsets for 24 h. The cardiac function tests were performed for all the patients 2 weeks after admission. Their echocardiographic parameters were analyzed, meanwhile the 24 h dynamic electrocardiogram detection was performed. The heart rate and the incidence of arrhythmia were recorded, and the incidences of heart failure and cardiogenic shock within the 2 weeks were observed and analyzed retrospectively. Results The experimental group showed that the cTnI level was (10.41±6.83) μg/L, BNP level was ( 1 713.6±687.03) ng/L, and heart rate was (86±12)/min, which were significantly higher than those in control group [(5.41±2.86) μg/L, (495.61±464.39) ng/L and (77±11)/min, respectively](P<0.05, P<0.01). The levels of BNP and cTnI had a positive correlation(r²=0.753, r=0.868, P<0.01). The differences on the incidence rates of malignant arrhythmia, heart failure and cardiogenic shock between experimental and control groups were statistically significant (P<0.05). In experimental group, the left ventricular ejection fraction(LVEF) was 48.83%±5.01%, and the left ventricular fractional shortening(LVFS) was 24.88%±6.53%, which were lower than those in control group (54.82%±5.29% and 30.86%±5.89%)(P<0.01). Results The BNP and cTnI levels can help for judging the severity of AMI patients with non-diabetic irritable hyperglycosemia and evaluate the recent prognosis. BNP and cTnI can be used as risk indicators for AMI patients with non-diabetic irritable hyperglycosemia.

Investigation on the correlation and clinical significance of serum mammaglobin and breast cancer

YANG Chao, KANG Wei, LI Jianhua, YOU Tao, WANG Yongfang.

2014, 29(1):  61-64.  DOI: 10.3969/j.issn.1673-8640.2014.01.015

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Objective To investigate the correlations of serum human mammaglobin(hMAM)levels with the early diagnosis of breast cancer and cancer micro-metastasis and their clinical significance. Methods By enzyme-linked immunosorbent assay(ELISA), the serum hMAM levels of 68 patients with breast cancer, 40 patients with other cancers (10 patients with prostate cancer, 10 patients with gastric cancer, 10 patients with ovarian cancer and 10 patients with colorectal cancer), 35 patients with benign breast disease and 40 healthy females (control group) were determined and analyzed comparatively. The 68 patients with breast cancer were classified according to TNM stage, whether existed the estrogen receptor (ER) expression, axillary lymph node metastasis and menopause. The cut-off value of serum hMAM in patients with breast cancer was determined by receiver operating characteristic (ROC) curve. Results The area under ROC curve of serum hMAM was 0.825, and the reliability in the diagnosis of breast cancer was 82.5% [95% confidence interval (CI): 72.6%-92.4%]. When the cut-off value for hMAM was 8.43 ng/mL, the sensitivity and specificity were 76.5% and 82.9%, respectively. Serum hMAM levels and positive rate in breast cancer group were significantly higher than those in benign breast disease group, other cancer group and control group (P<0.05). The difference among benign breast disease group, other cancer group and control group was not statistically significant (P>0.05). The positive rates of serum hMAM of Ⅲ and Ⅳ breast cancer patients (55% and 75%) were significantly higher than those of Ⅰ and Ⅱ patients (25% and 40%, P<0.05), but hMAM levels had no difference (P>0.05). The positive rate of serum hMAM in breast cancer patients with axillary lymph node metastasis (88%) was significantly higher than those without metastasis (30%, P<0.05), but hMAM levels had no difference (P>0.05). Regardless of whether existed ER expression, proto-oncogene C-erb-2 expression and menopause, serum hMAM levels and positive rate of patients with breast cancer had no difference (P>0.05). Results The hMAM expression is related with the clinical stage and axillary lymph node metastasis of breast cancer. The determination of serum hMAM level is helpful to improve the early diagnosis of breast cancer and early detection of cancer micro-metastasis.

The phenotypic detection of metallo-beta-lactamase-producing Pseudomonas aeruginosa by modified imipenem-EDTA disk potentiation test

YAN Yuzhong, FAN Huiqing, ZHENG Wenlong, YANG Huanzhang, LU Yanchun, SHI Yi.

2014, 29(1):  65-68.  DOI: 10.3969/j.issn.1673-8640.2014.01.016

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Objective To evaluate the modified imipenem-ethylene diamine tetraacetic acid (EDTA) disk potentiation test for the phenotypic detection of metallo-beta-lactamase-producing Pseudomonas aeruginosa(PA). Methods A total of 95 carbapenem-resistant PA were collected. The minimum inhibition concentrations(MIC) of imipenem and meropenem were determined by agar dilution method.Polymerase chain reaction (PCR) and DNA sequencing were used for the identification of metallo-beta-lactamase genotypes. The phenotypic detection of metallo-beta-lactamase was done by conventional and modified imipenem-EDTA disk potentiation test. Results The resistance rates of imipenem and meropenem were 100.0% and 71.6% among these 95 carbapenem-resistant PA. The sensitivity, specificity, positive predictive value and negative predictive value of conventional test were 100.0%, 96.6%, 72.7% and 100.0%, respectively. Those of modified test were all 100.0%. Results The modified imipenem-EDTA disk potentiation test could be an effective method for the phenotypic detection of metallo-beta-lactamase-producing PA.

The determination of 5 amino acid neurotransmitters in hippocampus of rats by RP-HPLC with fluorescence detector

BAI Yu, ZHANG Li, MA Xiaoli, KONG Bin, LI Xinxia, LI Xinyu.

2014, 29(1):  69-72.  DOI: 10.3969/j.issn.1673-8640.2014.01.017

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Objective To establish a method of reverse phase high performance liquid chromatography(RP-HPLC) with fluorescence detector to determine 5 amino acid [aspartate (Asp), glutamate(Glu), asparagine (Aspa), glutamine (Gln) and glycine (Gly)] neurotransmitters. Methods The amino acid neurotransmitters in hippocampus of rats were collected by methanol: water=1∶1. The RP-HPLC with fluorescence detector with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) pre-column derivatization was used to measure the 5 amino acid neurotransmitters in hippocampus of rats. Results The 5 amino acid neurotransmitters were well and completely separated using elution within 15 min. There was a good linear correlation in the range of 0.5-100.0 μg/μL, and the correlation coefficient (r) was≥0. 999 90. The detection limits of Asp, Glu, Aspa, Gln and Gly were 0.05, 0.068, 0.037, 0.05 and 0.02 μg/μL. The precisions [relative standard deviation(RSD)] were 0.4%-2.5%. The recoveries of added amino acids were 70.27%-128.20%(RSD<5%).The contents of Asp, Glu, Aspa, Gln and Gly in hippocampus of normal rats were 38.98±9.66, 126.42±34.06, 216.00±0.75, 90.44±33.75 and (12.95±4.42) μg/g. Results RP-HPLC with fluorescence detector is quick, reliable and sensitive to measure the amino acid neurotransmitters in hippocampus of rats.

Research on advanced glycation end products in the mechanism of coronary artery disease

LIU Junfeng, KONG Meijuan, JIA Kegang.

2014, 29(1):  76-80.  DOI: 10.3969/j.issn.1673-8640.2014.01.019

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Advanced glycation end products (AGEs) are closely involved in the pathophysiological process of vascular smooth muscle cell (VSMCs) differentiation, proliferation, coronary artery disease and other cardiovascular diseases. AGEs can induce autophagy in VSMCs through the extracellular signal-regulated protein kinase(ERK) and serine-threonine kinase (Akt) signaling pathways. AGEs improve cardiac microvascular endothelial cell (CMECs) proliferation depending on bone marrow stromal cell-derived factor-1(SDF-1)/ chemokine receptor CXCR4 axis signaling pathway. AGEs-2 and AGEs-3 up-regulate the expression of receptor for advanced glycation end product (RAGE) on monocytes. AGEs inhibit the proliferation, migration and adhesion of endothelial progenitor cell (EPCs), and induce the apoptosis of EPCs. AGEs increase the expression activity of connective tissue growth factor(CTGF) mRNA and protein, and promote the proliferation of cardiac fibroblast and the secretion of transforming growth factor-beta 1 (TGF-β1) and induce the expressions of Smad2 and Smad4. The carboxymethyl lysine(CML)/RAGE axis plays an important role in atherosclerotic calcification of diabetes through the mechanism that induces the apoptosis of macrophages followed by the osteogenic differentiation of aortic smooth muscle cells. Soluble form of RAGE(sRAGE) can be served as bait of RAGE ligand to prevent atherosclerosis, and the sensitivity and negative predictive value of sRAGE are higher than those of AGEs/sRAGE ratio in identifying post-percutaneous coronary intervention (PCI) restenosis. ALT-711 is a breaker of AGEs-based cross links, which can inhibit AGEs-mediated formation of reactive oxygen species (ROS), extracellular signal-regulated kinase phosphorylation and cyclooxygenase-2 expression. Pigment epithelium-derived factor (PEDF) can inhibit CD40 ligand(CD40L) overexpression by blocking the effects of AGEs on platelets, which may become a therapeutic target for the prevention of coronary artery disease. Statins can also inhibit AGEs-induced aortic smooth muscle cell proliferation and production of ROS. The research above can reveal some the pathogenesis of coronary artery disease, provide foundation and direction for exploring the corresponding drug intervention and adjustment of clinical treatment strategies.

The introduction of evaluating measurement uncertainty with top-down approach

CHEN Longmei, WANG Huimin, JU Yi.

2014, 29(1):  81-85.  DOI: 10.3969/j.issn.1673-8640.2014.01.020

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The measurement of uncertainty is an important parameter which ensures the integrity of measurement results. Top-down approach is a prior choice to evaluate uncertainty in medical laboratories with imprecision and bias. Since 1995, when top-down approach was firstly introduced, the evaluation objects and calculating Methods have been developed and improved continuously. At present, imprecision can be determined by replicate measurements and internal quality control data, while bias can be assessed by certified reference materials, reference methods, recovery tests and external quality assessment data. The Methods of evaluating measurement uncertainty will be focused according to different objects in this review.

Common influence factors for the stability of urine storage

LUO Yi, WANG Congrong.

2014, 29(1):  86-90.  DOI: 10.3969/j.issn.1673-8640.2014.01.021

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Urine is easily accessible, can be collected noninvasively, and provides information on numerous physiological processes. Urine is a source of numerous potential biomarkers, including metabolites, cells, proteins and nucleic acids. Many factors can affect the quality of the stored urine, including the time and temperature of storage, PH and multiple freezing and thawing. However, it remains being a controversial topic on the influence of exterior factors on the stability of urine storage. With the development of proteomics and metabolomics, mounting number of metabolomic and proteomic studies attempt to identify the influence of storage conditions on urine analysis. This review presents an overview of the influence of storage conditions on urine analysis.