Table of Content

30 April 2014, Volume 29 Issue 4

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Orignal Article

Correlation research of HPV16E6 and hWAPL with cervical cancer

JING Fahong，MU Yudong，XU Jiru，KANG Wei

2014, 29(4):  303-306.  DOI: 10.3969/j.issn.1673-8640.2014.04.001

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Objective　To investigate the expressions of human papillomavirus 16E6 （HPV16 E6） and human wings apart-like gene （hWAPL） in different levels of cervical lesions and the relationship of HPV16E6 and hWAPL protein with cervical cancer， and to provide the reference for the early diagnosis of cervical cancer. Methods　By immunohistochemistry， the paraffin-embedded blocks with different levels of cervical lesions were detected， including 49 patients with cervical intraepitelial neoplasia （CIN） Ⅰ or CIN Ⅱ， 33 patients with CIN Ⅲ or carcinoma in situ， 28 patients with invasive cervical carcinoma （ICC） and 16 patients with normal cervical epithelium. Results　In normal， CINⅠ or Ⅱ， CIN Ⅲ and ICC groups， the positive expression rates of HPV16E6 and hWAPL showed gradually increasing trend. The positive expression rates of HPV16E6 and hWAPL in CIN Ⅲ and ICC groups had statistical significance with those in normal and CIN Ⅰ or CIN Ⅱ groups （P＜0.05）. Between CIN Ⅲ and ICC groups， the positive expression rate of HPV16E6 had statistical significance （P＜0.05）， but the positive expression rate of hWAPL protein had no obvious difference （P＝0.372）. HPV16E6 and hWAPL expression strengthes in ICC group and CIN Ⅲ group were obviously higher than those in CIN Ⅰ or Ⅱ group and normal group （P＜0.05）， and there was no obvious difference between ICC and CIN Ⅲ groups （P>0.05）. The 57 of 64 patients （89.06%） with HPV16E6 had the positive expression of hWAPL protein， 15 of 62 patients （24.19%） with no HPV16E6 had the positive expression of hWAPL protein. By statistical analysis， both HPV16E6 and hWAPL protein had significant correlation （P＜0.05）， and it showed that the infection intensity of HPV16E6 and the expression of hWAPL had positive correlation （r＝0.595， P＜0.001）. Conclusions　The hWAPL may be a cervical cancer-specific gene and expresses highly， and it may be important in the pathogenesis of cervical cancer. The hWAPL may become the new target gene for the diagnosis， treatment and lesion identification of cervical cancer.

Significance investigation of high-sensitivity cardiac troponin T for viral myocarditis in children

HU Xiaobin，XIANG Xiaojie，GUO Xiaocong，CHEN Huagang，FANG Hua，LU Xinmin.

2014, 29(4):  307-311.  DOI: 10.3969/j.issn.1673-8640.2014.04.002

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Objective　To investigate the diagnosis and treatment significance of high-sensitivity cardiac troponin T （hs-cTnT）and creatine kinase isoenzyme mass （CK-MB mass） for viral myocarditis in children （VMC）. Methods　Serum levels of hs-cTnT and CK-MB mass were determined in 67 patients with VMC （measured within clinical symptoms for 2- 4 d and after treatment for 1 week）， 54 children with acute gastroenteritis， 53 children with acute upper respiratory tract infection， 50 children with acute pneumonia and 55 healthy children （healthy control group）. The receiver operating characteristic （ROC） curve was used to evaluate the sensitivity and specificity of hs-cTnT determination， CK-MB mass determination and the combined determination of hs-cTnT and CK-MB mass. Results　Serum levels of hs-cTnT in acute gastroenteritis group， acute upper respiratory tract infection group， acute pneumonia group， healthy control group and VMC group [median （quartile）] were 5.68（3.83， 8.22）， 5.03（3.00， 6.57）， 5.52（3.44， 10.19）， 4.81（3.22， 6.67）and 58.57（29.77， 160.20）pg/mL， respectively， and serum levels of CK-MB mass [median （quartile）] were 2.29（1.72， 2.96）， 2.26（1.41， 3.48）， 2.59（1.81， 3.62）， 1.83（1.42， 3.05） and 7.40（4.32， 13.26） ng/mL， respectively. There was statistical significance for hs-cTnT and CK-MB mass among the groups （H=115.15 and 83.79，P<0.001）. Serum levels of hs-cTnT and CK-MB mass in VMC group after treatment were lower than those within clinical symptoms for 2- 4 d （P<0.001）， which were significantly higher than those in the acute gastroenteritis group， acute upper respiratory tract infection group， acute pneumonia group and healthy control group （P<0.001）. The ROC curve showed that the combined determination of hs-cTnT and CK-MB mass had higher sensitivity than hs-cTnT determination （P<0.05）， but without significant specificity （P>0.05）. Conclusions　The hs-cTnT has high diagnosis sensitivity and specificity for VMC， and the combined determination of hs-cTnT and CK-MB mass has high diagnosis significance.

The clinical significance on the combined determination of NT-proBNP, H-FABP and cTnI in elder patients with severe heart failure

WU Shuzhi，DENG Sheng，QIN Weiguo，CHEN Jian，YI Bin.

2014, 29(4):  312-318.  DOI: 10.3969/j.issn.1673-8640.2014.04.003

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Objective　To study the clinical significance on the combined determination of serum N-terminal pro-B-type natriuretic peptide (NT-proBNP), heart-type fatty acid-binding protein(H-FABP)and cardiac troponin I(cTnI)in elder patients with severe heart failure (HF). Methods　The serum NT-proBNP, H-FABP and cTnI levels were determined in 312 elder patients with severe HF ｛main adverse cardiovascular event (MACE)group(194 cases)[cardiogenic readmission group(110 cases) and death group(84 cases)]and no-MACE group(118 cases)｝on admission, day 7, 15, 30 and 60 after treatment, 76 elder patients with no-severe HF and 40 elder healthy subjects. The MACE were followed and observed for 180 d. The significance of NT-proBNP, H-FABP and cTnI for the diagnosis of severe HF and the prediction of death risk was analyzed by receiver operating characteristic(ROC)curve. Survival analysis was performed by Kaplan-Meier curve. Results　The serum NT-proBNP, H-FABP and cTnI levels in elder patients with severe HF were significantly higher than those in elder patients with no-severe HF and healthy subjects (P<0.01), the positive rate of H-FABP in elder patients with no-severe HF was 47.1％, and the positive rate of cTnI was 4.5％. The serum NT-proBNP levels were positively correlated with the grade of New York Heart Association (NYHA)Ⅰ-Ⅳ (r=0.77,P<0.01). The ROC curve indicated that the optimal cut-off values of serum NT-proBNP,H-FABP and cTnI levels for diagnosis in elder patients with severe HF were 4 601.50 ng/L, 5.16 ng/L and 0.025 ng/mL, and the optimal cut-off values of serum NT-proBNP,H-FABP and cTnI levels for the death risk evaluation in elder patients with severe HF were 8 178.50 ng/L, 11.77ng/L and 0.038ng/mL, respectively. The sensitivity of the combined determination was obviously better than those of the individual determinations (P<0.05). Before the treatment, the serum NT-proBNP levels in MACE group were significantly higher than those in no-MACE group(P<0.05), the serum H-FABP levels in death group were significantly higher than those in no-death group(P<0.05), and the serum cTnI levels in various prognosis groups had no statistical significance (P>0.05). The serum NT-proBNP, H-FABP and cTnI levels in no-MACE group on day 7 after treatment were significantly lower than those before the treatment (P<0.05). The decreasing rates of the 3 parameters were >50%, and the serum H-FABP level was lower than the normal reference value. The serum NT-proBNP, H-FABP and cTnI levels in the cardiogenic readmission group on day 15 after treatment were significantly lower than those before treatment(P<0.05), but the decreasing rates of the 3 parameters were <30%,and the serum H-FABP level was higher than the normal reference value. The serum NT-proBNP, H-FABP and cTnI levels in death group on day 7 and day 30 after treatment were significantly higher than those before treatment(P<0.05), and the serum cTnI levels were higher than the normal reference value. Kaplan-Meier survival curve showed that the survival rates of serum NT-proBNP, H-FABP and cTnI levels had significant difference(P<0.05). Conclusions　There is an important clinical significance of the combined determination of serum NT-proBNP, H-FABP and cTnI levels for the diagnosis, prognosis, death risk evaluation and curative effect survey in elder patients with severe HF.

Research on the cut-off value of microalbuminuria among type 2 diabetes mellitus

LI Rong，WANG Huabin，LIU Rui，CUI Xiaofan，PAN Wenjie

2014, 29(4):  319-323.  DOI: 10.3969/j.issn.1673-8640.2014.04.004

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Objective　To research the cut-off value of microalbuminuria with urine albumin to creatinine ratio（ACR） and urinary albumin（U-Alb） concentration of first morning urine and random urine samples among type 2 diabetes mellitus（T2DM）， in order to detect early diabetic kidney disease（DKD）. Methods　A total of 169 T2DM patients and 40 healthy subjects（healthy control group） were enrolled. Their 24 h urine， first morning urine and random urine samples were collected. The 24 h urinary albumin excretion （UAE） was referred as the gold standard of early DKD， and the detection effectiveness of early DKD was evaluated when applied ACR and U-Alb concentration from first morning urine and random urine samples. According to receiver operating characteristic （ROC） curve analysis， the value with the maximum Youden index was the appropriate microalbuminuria cut-off value of ACR and U-Alb concentration from first morning urine and random urine samples for detecting early DKD. Results　The coincidence of ACR and U-Alb concentration with 24 h UAE from first morning urine samples were 43% and 37%， and those of ACR and U-Alb concentration from random urine samples were 48% and 41%， which were all significantly different with 24 h UAE（P＜0.001）. In healthy control group， the microalbuminuria judgment was totally normal and consistent with 24 h UAE. ROC curve analysis indicated that the cut-off values of ACR from first morning urine samples were male 16 mg/g and female 23 mg/g （Youden indices 0.7 and 0.67， negative predictive values 97% and 100%， and positive predictive values 72% and 65%， respestively）， and those of ACR from random urine samples were male 17 mg/g and female 28 mg/g（Youden indices 0.68 and 0.67， negative predictive values 90% and 90% and positive predictive values 61% and 82%， respectively）. Those of U-Alb concentration from first morning urine samples were male 16 mg/L and female 15 mg/L（Youden indices 0.57 and 0.59， negative predictive values 84% and 87%， and positive predictive values 90% and 73%， respectively）， and those of U-Alb concentration from random urine samples male 17 mg/L and female 14 mg/L（Youden indices 0.56 and 0.53， negative predictive values 73% and 81%， and positive predictive values 85% and 71%， respectively）. The maximum Youden index of ACR was >0.6 and better than the Youden index of U-Alb concentration. Conclusions　The missing diagnosis rate is high when detecting early DKD among T2DM patients with current clinical microalbuminuria cut-off value from first morning urine and random urine samples. The cut-off values of ACR and U-Alb concentration from first morning urine and random samples should be established in order to facilitate the early prevention and treatment of DKD.

Investigation on the correlation between glycosylated hemoglobin A_1c and biochemical markers of bone metabolism

ZHANG Min, LI Zhi, WEI Qi, WANG Lu

2014, 29(4):  324-326.  DOI: 10.3969/j.issn.1673-8640.2014.04.005

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Objective　To research the cut-off value of microalbuminuria with urine albumin to creatinine ratio（ACR） and urinary albumin（U-Alb） concentration of first morning urine and random urine samples among type 2 diabetes mellitus（T2DM）， in order to detect early diabetic kidney disease（DKD）. Methods　A total of 169 T2DM patients and 40 healthy subjects（healthy control group） were enrolled. Their 24 h urine， first morning urine and random urine samples were collected. The 24 h urinary albumin excretion （UAE） was referred as the gold standard of early DKD， and the detection effectiveness of early DKD was evaluated when applied ACR and U-Alb concentration from first morning urine and random urine samples. According to receiver operating characteristic （ROC） curve analysis， the value with the maximum Youden index was the appropriate microalbuminuria cut-off value of ACR and U-Alb concentration from first morning urine and random urine samples for detecting early DKD. Results　The coincidence of ACR and U-Alb concentration with 24 h UAE from first morning urine samples were 43% and 37%， and those of ACR and U-Alb concentration from random urine samples were 48% and 41%， which were all significantly different with 24 h UAE（P＜0.001）. In healthy control group， the microalbuminuria judgment was totally normal and consistent with 24 h UAE. ROC curve analysis indicated that the cut-off values of ACR from first morning urine samples were male 16 mg/g and female 23 mg/g （Youden indices 0.7 and 0.67， negative predictive values 97% and 100%， and positive predictive values 72% and 65%， respestively）， and those of ACR from random urine samples were male 17 mg/g and female 28 mg/g（Youden indices 0.68 and 0.67， negative predictive values 90% and 90% and positive predictive values 61% and 82%， respectively）. Those of U-Alb concentration from first morning urine samples were male 16 mg/L and female 15 mg/L（Youden indices 0.57 and 0.59， negative predictive values 84% and 87%， and positive predictive values 90% and 73%， respectively）， and those of U-Alb concentration from random urine samples male 17 mg/L and female 14 mg/L（Youden indices 0.56 and 0.53， negative predictive values 73% and 81%， and positive predictive values 85% and 71%， respectively）. The maximum Youden index of ACR was >0.6 and better than the Youden index of U-Alb concentration. Conclusions　The missing diagnosis rate is high when detecting early DKD among T2DM patients with current clinical microalbuminuria cut-off value from first morning urine and random urine samples. The cut-off values of ACR and U-Alb concentration from first morning urine and random samples should be established in order to facilitate the early prevention and treatment of DKD.

The changes of blood coagulation and fibrinolytic indices after radiofrequency catheter ablation

CUI Fan

2014, 29(4):  328-330.  DOI: 10.3969/j.issn.1673-8640.2014.04.007

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Objective　To observe the changes of blood coagulation and fibrinolytic indices after radiofrequency catheter ablation(RFCA) and the restoring time after RFCA. Methods　The levels of D-dimer (DD), von willebrand factor (vWF), tissue plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI-1) were determined in 56 patients undergoing RFCA after intracardiac electrophysiological study before RFCA, just at the time after RFCA and in 2 d and 7 d after RFCA. Results　The levels of DD, vWF and PAI-1 increased significantly after intracardiac electrophysiological study before RFCA, just at the time after RFCA and in 2 d after RFCA (P<0.01), and restored in 7 d after RFCA (P>0.05). The t-PA decreased significantly after intracardiac electrophysiological study before RFCA, just at the time after RFCA and in 2 d after RFCA (P<0.01), and restored in 7 d after RFCA (P>0.05). Conclusions　RFCA may increase the levels of blood coagulation and fibrinolytic indices. Monitoring the levels of D-dimer, vWF, t-PA and PAI-1 is valuable in guiding the administration of anticoagulative drugs and the prophylaxis of thromboembolic events.

The significance of fibronectin detection in hepatofibrosis

YANG Jingjing，JIANG Hongmin，TANG Aiguo.

2014, 29(4):  331-336.  DOI: 10.3969/j.issn.1673-8640.2014.04.008

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Objective　To investigate the change of serum fibronectin （Fn） level in patients with hepatopathy and the significance of Fn in hepatofibrosis. Methods　Serum Fn levels in patients with hepatopathy and healthy controls were determined. Serum total bile acid （TBA） and albumin（Alb） were determined， and the results were analyzed comparatively. Results　Serum Fn levels in patients with oxyhepatitis and liver cancer had no statistical significance with those in healthy controls with statistical significance（P>0.05）. Serum Fn levels in patients with chronic moderate hepatitis， chronic severe hepatitis and cirrhosis were lower than those in healthy controls with statistical significance（P<0.05）. Serum Fn levels in patients with cirrhosis were lower than those in other patients with hepatopathy（P<0.05）. Serum Fn levels in patients with alcohol cirrhosis and virus cirrhosis had no statistical significance（P>0.05）. Serum Fn levels in cirrhosis patients with complications，such as peritonitis， hemorrhage of digestive tract and hypersplenia， decreased significantly than those without complications（P<0.05）. Serum Fn and TBA in healthy controls and patients with oxyhepatitis and liver cancer had no correlation （P>0.05）， and had negative correlation between patients with chronic moderate hepatitis， chronic severe hepatitis and cirrhosis（r=-0.593，－0.677 and －0.714， P<0.05）. Serum Fn had no correlation with Alb （P>0.05）. Conclusions　Serum Fn can reflect the function of hepatocyte in a certain extent， and has certain reference significance in the diagnosis of hepatofibrosis especially cirrhosis and cirrhosis with infection. The diagnostic accuracy and reliability of Fn determination with TBA and Alb can be obviously improved.

Evaluation on the "WBC Correct" function performance of BECKMAN-COULTER LH750 hematology system

FANG Meirong，CHEN Xiaoli

2014, 29(4):  337-338.  DOI: 10.3969/j.issn.1673-8640.2014.04.009

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Objective　To analyze statistically the accuracy and reliability of a special function for white blood cell （WBC） count in BECKMAN-COULTER LH750 hematology system （LH750）——"WBC Correct" function. Methods　According to the Reference Method for Red Cell and White Cell Counting published by the Ministry of Health， the WBC counts of the samples corrected automatically by machine had been counted manually. The results were analyzed statistically. Results　There was a good correlation for the results by LH750 WBC Correct function and manual counting method， and LH750 WBC Correct function had good accuracy and reliability. There were 5 wrong corrections among 420 samples. Conclusions　The LH750 WBC Correct function is practically significant. However， there are some wrong corrections. Therefore， it should be paid more attention to the correction flags. Manual counting method should be carried on when a wrong correction occurs.

Laboratory diagnosis research of AIDS combined with penicilliosis marneffei

WEI Shanqiu, SU Guosheng, LAN Jiang, LUO Xiaolu.

2014, 29(4):  339-341.  DOI: 10.3969/j.issn.1673-8640.2014.04.010

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Objective　To investigate the laboratory diagnosis Methods　of acquired immune deficiency syndrome（AIDS） combined with penicilliosis marneffei（PSM）. Methods　A total of 257 AIDS patients were enrolled. Bone marrow and peripheral blood fungal dimorphic culture and bone marrow smear microscopy were performed for the detection of Penicillium marneffei（PM）. The data were analyzed retrospectively. Results　The 77 AIDS patients had PM， and the detection rate was 29.96%. A total of 74 patients were identified by bone marrow and peripheral blood fungal dimorphic culture， including 65 patients were positive in bone marrow， and 54 patients were positive in peripheral blood. A total of 33 patients were proved to be PM positive by bone marrow smear microscopy， including 30 patients showed positive in fungal dimorphic culture， and 3 patients showed negative but positive during therapeutic diagnosis. With the positivity of bone marrow fungal and peripheral blood fungal dimorphic culture as the good standard for PM diagnosis， the detection rate of PM by bone marrow fungal culture was 87.84%， which was significantly higher than the detection rate by peripheral blood fungal culture among AIDS patients（72.97%）. The 2 individual culture rates were significant statistically with the rate of dimorphic culture（P＜0.05）. The detection specificity for the diagnosis of PSM by bone marrow smear microscopy was 90.91%， and the sensitivity was 44.59%. Combined with bone marrow smear microscopy， the missing rate of dimorphic culture was 3.90%. Conclusions　The detection of PM by bone marrow and peripheral blood fungal dimorphic culture combined with bone marrow smear microscopy is significant in the diagnosis of AIDS combined with PM on time.

Experimental research of honeysuckles and other six kinds of traditional Chinese medicine air spray to reduce nosocomial infection rate

LU Meizhu, HUANG Caijun, GE Kangyan.

2014, 29(4):  342-346.  DOI: 10.3969/j.issn.1673-8640.2014.04.011

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Objective　To observe the effects of honeysuckles and other 6 kinds of traditional Chinese medicine air spray　to control the nosocomial infection. Methods　Spraying the prepared honeysuckles and other 6 kinds of traditional Chinese medicine on anion humidifier， the air bacterium content was observed before and after spraying. The infection amount of all the relevant hospitalized patients and the use of antimicrobial drugs were analyzed. Results　Compared with that before spray sterilization， the number of colony after spray sterilization decreased significantly（P＜0.05）， and the nosocomial infection rate also decreased（P＜0.05）. The usage rate of antimicrobial drugs decreased in different degrees. Conclusions　The application of honeysuckles and other traditional Chinese medicine air spray is able to achieve the purpose of reducing nosocomial infection rate.

Significance investigation of applying high risk-type HPV DNA detection in healthy examination

ZHU Rong，JU Huixiang，WANG Feng，LU Qiang

2014, 29(4):  347-349.  DOI: 10.3969/j.issn.1673-8640.2014.04.012

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Objective　To investigate the clinical application significance of high risk-type human papillomavirus（HPV） DNA detection to screen cervix precancerous lesion in healthy examination. Methods　A total of 2 827 married females for healthy examination did the common gynecological examination. Their samples were detected for thinprep cytology test（TCT） and HPV 13 high risk-type DNA. Results　The infection rate of high risk-type HPV was 15.9%， and the infection rates of high risk-type HPV in 3 different groups（＜30 years old， 30-45 years old and ＞45 years old） had no statistical significance（P＞0.05）. The 83.4％ females who had high risk-type HPV infection could remove virus infection in 1 year， and 372 females who had no high risk-type HPV infection did not have precancerous lesion or cervical cancer at least 2 years. The infection rate of high risk-type HPV in 2 groups of cervix slick or low-degree erosion and cervical erosionⅡor above degree had statistical significance（P＜0.05）. The screen rate of cervix precancerous lesion and cervical cancer using TCT and high risk-type HPV DNA combination detection was 72.7%. Conclusions　The application of high risk-type HPV DNA detection in healthy examination has early diagnosis， prevention and treatment significance in the negative prediction of high risk-type HPV infection and persistent infection of HPV above 12 months. TCT and high risk-type HPV DNA combination detection can screen cervical cancer and cervix precancerous lesion efficiently.

The preliminary evaluation of multiple genetic analysis system to detect Helicobacter pylori

ZHOU Lifang, BAO Zhijun,MIAO Yingxin,JI Danian,HUANG Yiqin,ZHAO Fuju,ZHAO Hu.

2014, 29(4):  350-356.  DOI: 10.3969/j.issn.1673-8640.2014.04.013

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Objective　To evaluate a new kind of Helicobacter pylori（HP） detection method， using the multiple genetic analysis system（GeXP system） to detect HP infection. Methods　Use culture method， rapid urease test， ordinary polymerase chain reaction（PCR） and multiple genetic testing to determine whether gastric biopsies exist HP. The sensitivity， specificity， positive predictive value and negative predictive value were evaluated by Stata 12.0 statistical software， in order to evaluate the testing performance of GeXP system from horizontal degree. Furthermore， multiple genetic testing was performed by selecting 5 specific genes（16S rRNA, ureA, ureC, 26KDa and cagA genes）as HP diagnosis infection， and multiple genetic testing was evaluated from longitudinal degree. Results　The sensitivity of culture method was 76.8%， the specificity was 100.0%， the positive predictive value was 100.0%， and the negative predictive value was 57.7%. For rapid urease test， by contrast， its sensitivity was 87.8%， the specificity was 80.8%， the positive predictive value was 93.5%， and the negative predictive value was 67.7%. Ordinary PCR had the highest sensitivity and specificity（100.0%）. For GeXP system， the sensitivity was 100.0%， the specificity was 71.3% without false negative case， the positive predictive value was 90.9%， and the negative predictive value was 100.0%. Conclusions　The new preliminary multiple genetic testing， GeXP system， has high sensitivity and specificity and high flux characteristics， which would not only meet the needs of clinical sample determination， but also could monitor prognosis after receiving eradication therapy. The GeXP system has a promising application significance.

The induction of vascular endothelial microparticles to T cell activation and the cytokine production of Th1 type

LIU Changying，SUN Yi

2014, 29(4):  357-365.  DOI: 10.3969/j.issn.1673-8640.2014.04.014

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Objective　To study the influence of vascular endothelial microparticles（EMP） on T lymphocyte activation in vitro and the cytokine production， and to study the roles of EMP on immune response. Methods　Lymphocytes from BN rats were interfered by EMP derived from human endothelial cells and SD rat′s endothelial cells. The early activation CD69 expression of T lymphocytes（CD3⁺ lymphocytes） was measured by flow cytometry. Cytokines such as interferon-gamma（IFN-γ） and interleukin 2（IL-2） were measured for expression by enzyme-linked immunosorbent assay（ELISA）. Results　Compared with those of control group [adding same volune of phosphate buffer（PBS）]， the expressions of CD69 and IFN-γ in culture supernatant of each EMP concentration group increased remarkably（P＜0.05）. Compared with those of control group， the expression of IL-2 increased remarkably in culture supernatant of intermediate and high EMP concentration groups with statistical significance（P＜0.05）. Conclusions　EMP can induce T lymphocyte activation and Th1 cytokine production. EMP may play an important role in immune response through activating T lymphocytes.

Research on the mechanisms of streptococcal protein inducing cycle arrest and apoptosis in glioblastoma BT325 cell

WANG Yan，ZHANG Liping，CHEN Hui，CAO Ning，ZHU Junping，PING Guoling，LI Weihong

2014, 29(4):  363-368.  DOI: 10.3969/j.issn.1673-8640.2014.04.015

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Objective　To investigate the mechanisms of streptococcal protein inducing cycle arrest and apoptosis in human cerebral glioblastoma BT325 cell. Methods　Human cerebral glioblastoma BT325 cells were cultured in vitro， and methlcyclopentadienyl manganese tricarbonyl（MTT） assay was used to determine the proliferation activity of BT325 cells. The morphological changes of BT325 cells were observed by the phase-contrast microscopy. The apoptotic cells were observed after fluorescence staining. Flow cytometry was used to determine the cell cycle， the rate of cell apoptosis and the change of mitochondrial membrane potential（MMP）. The expression of apoptosis-related proteins was determined by Western blot. Results　Streptococcal protein（10-100 mg/L） inhibited the proliferation of BT325 cells significantly and induced G2/M phase arrest in time- and dose-dependent manners. MMP declined（P＜0.01）， and apoptotic cells gradually increased after BT325 cells were treated with streptococcal protein for 24 h. The rate of cell apoptosis in determination group was significantly higher than that in control group at 48h（P＜0.01）. In a time-dependent manner， streptococcal protein（50 mg/L） up-regulated the expression of cell P53 protein， down-regulated the expression of B-cell lymphoma-2（Bcl-2）， led to the release of mitochondrial cytochrome C（CytC） from mitochondria into cytoplasm， and then caused the activation of the precursor of cysteinyl aspartate specific proteinase 3（Procaspase-3）. Conclusions　Streptococcal protein may exert its anticancer activity through inhibiting the proliferation of human cerebral glioblastoma BT325 cells， effecting the distribution of cell cycle and inducing cell apoptosis by mitochondrial-dependent pathway.

Study on the resistance mechanism of an isolate of Klebsiella pneumoniae resistant to 3 kinds of carbopenems

XIE Chunbao, YU Hua, XIAO Daiwen, YANG Yongchang, JIANG Wei, LIU Hua, HUANG Wenfang.

2014, 29(4):  369-374.  DOI: 10.3969/j.issn.1673-8640.2014.04.016

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Objective　To investigate the resistance mechanism of an isolate of Klebsiella pneumoniae（K30） resistant to carbopenems. Methods　Minimum inhibition concentrations（MIC） of Klebsiella pneumoniae K30 to 13 antibiotics were determined by agar dilution method. Modified Hodge test was used to detect carbopenems. Class A carbopenem（KPC）， Class B carbopenem（NDM, IMP, VIM and SIM）， extended spectrum beta-lactamases [ESBLs（CTX, TEM and SHV）]， AmpC beta-lactamases [Amp C（FOX, EBC, ACC, DHA, CIT and MOX）] and Class Ⅰ integron were amplified by polymerase chain reaction（PCR）. Real-time fluorescence quantitation PCR was carried out to investigate the mRNA expression levels of porin genes（ompK35 and ompK36）. Plasmid conjugation experiment was subjected to reveal the transferability of Klebsiella pneumoniae K30 resistant genes. Results　The antimicrobial susceptibility test showed that Klebsiella pneumoniae K30 was resistant to 11 antibiotics， but kept intermediary to cefoxitin sodium and susceptible to amikacin. Modified Hodge test was positive in Klebsiella pneumoniae K30. Class A carbopenem KPC gene and 2 ESBLs CTX and SHV genes were positive by PCR amplification. The genes were conformed as KPC-2, CTX-M3 and SHV-38 by sequencing and comparing in GenBank. No other resistance gene Class Ⅰ integron was detected. The porin gene ompK35 and ompK36 expression levels did not decrease comparing with those of Klebsiella pneumoniae（ATCC 700603）. Plasmid conjugation experiment was failed in the study. Conclusions　The main resistance mechanism of the isolate of Klebsiella pneumoniae resistant to carbopenems might be associated with the combined producing KPC-2 and ESBLs， and KPC-2 may be not mediated by plasmid.

A clinical isolate of carbapenem-resistant Enterobacter aerogen with plasmid-mediated KPC-2 and DHA-1 genes

KUAI Shougang，WANG Weiping，PEI Hao，FAN Ming，LIU Jun，ZHOU Xike，SHANG Zhongbo，SHAO Haifeng

2014, 29(4):  375-379.  DOI: 10.3969/j.issn.1673-8640.2014.04.017

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Objective　To investigate the resistance and transmission mechanisms of a clinical isolate of carbapenem-resistant Enterobacter aerogen. Methods　The minimal inhibition concentrations（MIC） of antimicrobial agents were determined by agar dilution method， and plasmid conjugation experiment， plasmid extraction and DNA molecular hybridization， isoelectric focusing electrophoresis（IEF）， polymerase chain reaction（PCR）， DNA sequencing and outer-membrane protein analysis were used for analyzing the resistant gene and transmission mechanism. Results IEF showed that there were 3 bands of beta-lactamases with isoelectric point（pI） of 5.4， 6.7 and 7.8 in the clinical isolate of carbapenem-resistant Enterobacter aerogen. These 3 bands of beta-lactamases were confirmed to be TEM-1（pI 5.4）， KPC-2（pI 6.7） and DHA-1（pI 7.8） by PCR amplification and DNA sequencing， and 2 of them（pIs of 6.7 and 7.8） were found that they can be transferred by plasmid conjugantion experiment. KPC-2 and DHA-1 genes were located on an about 56 kb plasmid by plasmid conjugation experiment， plasmid extraction and DNA molecular hybridization. Outer-membrane protein electrophoresis analysis revealed that a 41 000 outer-membrane protein was absent in the clinical isolate of carbapenem-resistant Enterobacter aerogen comparing with clinical wild-type Enterobacter aerogen. Conclusions The clinical isolate of Enterobacter aerogen resistant to carbapenem produces a plasmid-mediaed carbapemase KPC-2 which belongs to Group 2f， Class A beta-lactamase. DHA-1 enzyme with outer-membrane protein absence may be related with the resistant mechanism of carbapenem-resistance in the isolate of Enterobacter aerogen.

A study on the establishment of the mechanism for tracing and warning the critical value reporting under the background of information management

SUN Jie，XU Weixin，REN Jun

2014, 29(4):  380-383.  DOI: 10.3969/j.issn.1673-8640.2014.04.018

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Objective　To use laboratory information management system（LIS）， hospital information management system（HIS） and mobile phone short message service（SMS） into becoming a comprehensive information technology platform， to establish the mechanism of tracing and warning the critical value reporting， and to enhance the significance of critical value reporting for the clinical practice. Methods　The data of information technology platform for improving critical value reporting were analyzed statistically. The clinical utility of the key mechanism for tracing and warning the critical value reporting was analyzed. Results　The critical value on the information technology platform for critical value reporting improved the timeliness and accuracy， and for clinical doctors， won more valuable time for the treatment of critical sickness patients. The establishment of the mechanism for tracing and warning the critical value reporting had the significance for the clinical practice. Conclusions　Critical value reporting realizes the establishment of information management and key mechanism for tracing and warning， and strengthens the critical value reporting significance for early warning， in order to improve emergency treatment efficiency for clinical doctors among critical sickness patients and prevent the related medical risk as a positive significance. It increases effectively the quality of patient′s treatment.

An example of MDS transformed into AML-M0

LAN Haijian，XIE Shuxia，ZHAO Wenhua，MENG Na，YU Jiuxiu

2014, 29(4):  384-386.  DOI: 10.3969/j.issn.1673-8640.2014.04.019

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Objective　To provide realistic reference for the diagnosis of minimal differentiation type of acute myelocytic leukemia（AML-M0）. Methods　The clinical process of a case of myelo-dysplastic syndrome（MDS） transformed into AML-M0 was reported. Results　Although MDS was performed for effective treatment， the most were still transforming into leukemia， mainly AML. Conclusions　The diagnosis of AML-M0 based on cell morphology and chemical staining must use the cell immunophenotype analysis and other modern means.

The research development of miRNA for the early detection of pancreatic cancer

LIAN Mi, MI Xianqiang.

2014, 29(4):  395-399.  DOI: 10.3969/j.issn.1673-8640.2014.04.023

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Pancreatic cancer is a malignant tumor with poor prognosis and extremely high mortality. Its diagnosis mainly relies on clinical imaging. Current diagnostic kits for pancreatic cancer diagnosis have the disadvantages of low specificity and accuracy, which results the poor early detection and diagnosis. In recent years, molecular diagnostics has been one of newly developing clinical diagnostic means. miRNA is abnormally expressed in pancreatic cancer, and it can become a novel tumor marker. The miRNA detection technology has developed continuously. This paper aims to review the current clinical diagnosis and diagnostic markers for pancreatic cancer and explore some methods used in the early detection of miRNA for pancreatic cancer.