Table of Content

30 January 2013, Volume 28 Issue 1

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Study on in vitro antibacterial activity of the combined therapy of imipenem with polymyxin B against Pseudomonas aeruginosa with different phenotypes

YANG Haihui 1，HAN Lizhong 2，LIU Yijing 1，CHEN Xu 2，YING Chunmei 1，ZHAO Beidi 1，ZHU Yanping 1，WEI Yingjue 1

2013, 28(1):  1-6.  DOI: 10.3969/j.issn.1673-8640.2013.01.001

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Objective　To investigate the in vitro antibacterial activity of the combined therapy of imipenem with polymyxin B against Pseudomonas aeruginosa with different phenotypes.   Methods　A total of 133 clinically isolated Pseudomonas aeruginosa were collected from Renji Hospital（West Department）and Ruijin Hospital，and the phenotypes were determined by disc diffusion（K-B）method. The minimum inhibitory concentrations（MIC）of imipenem，polymyxin B and imipenem with polymyxin B against Pseudomonas aeruginosa were calculated by agar dilution method，and the fractional inhibitory concentrations（FIC）were calculated. The combined effect was analyzed. There were 2 ratios，1∶0.5 and 1∶1 with imipenem to polymyxin B.  Results　The drug sensitivities of those 2 therapies turned out to be inconsistent when conducting Kappa test. The synergistic rate of 1∶1 with imipenem to polymyxin B was 51.13%，which was much higher than 1∶0.5 with imipenem to polymyxin（19.55%）. Fisher′s exact test demonstrated that 1∶1 with imipenem to polymyxin B maintained high synergistic rate between different phenotypes，and there was no statistically significant difference. However，the difference was significant regarding to 1∶0.5 with imipenem to polymyxin B.  Conclusions　Comparing with monotherapy，the combined therapy of imipenem with polymyxin B significantly increases the sensitivity of Pseudomonas aeruginosa in vitro. The combined therapy applying 1∶1 with imipenem to polymyxin B with a higher synergistic rate than that applying 1∶0.5 with imipenem to polymyxin B and a good stability may be a good choice for the antibiotics resistant Pseudomonas aeruginosa with different phenotypes.

Significance of plasma 1，3-beta-D-glucan assay for the diagnosis of invasive fungal diseases in patients with malignant hematological diseases

CHEN Yan 1，WANG Shuhui 2，HUANG Baoqing 2，NI Xiong 3，HUANG Xiaochun 1，QIN Yanghua 1

2013, 28(1):  7-10.  DOI: 10.3969/j.issn.1673-8640.2013.01.002

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Objective　To evaluate the significance of plasma 1，3-beta-D-glucan assay(G test)for the diagnosis of invasive fungal diseases（IFD）in patients with malignant hematological diseases. Methods　A retrospective analysis was performed in 220 inpatients with malignant hematological trmor．According to Diagnostic Criteria and Principles of Treatment of Invasive Fungal Infections in Patients with Blood Diseases/Malignancies as gold standard，the plasma 1，3-beta-D-glucan concentration′s area under the receiver operating characteristic（ROC）curve，sensitivity，specificity and time to diagnosis in the diagnosis of IFD were analyzed. Results　Comparing the gold standard，the plasma 1，3-beta-D-glucan concentration′s area under the ROC curve，sensitivity，specificity and time to diagnosis were 0.946 1，98.6%，83.0% and（3.86±2.92)d，respectively. Conclusions　G test has good sensitivity and time to diagnosis for the diagnosis of IFD in patients with malignant hematological diseases，and has a good negative predictive value in early diagnosis and exclusion of IFD. It can improve the efficiency and reduce the care burden of patient.

Analysis on Mycoplasma pneumonia 23SrRNA gene mutation site and drug resistance phenotype

YE Yun，LI Suliang，JIANG Ping，WANG Yao，YANG Chao

2013, 28(1):  11-16.  DOI: 10.3969/j.issn.1673-8640.2013.01.003

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Objective　To investigate the infection situation of Mycoplasma pneumonia（Mp）in patients with community-acquired respiratory tract infection and the molecular drug resistance mechanisms of macrolide，and to analyze the relationship between 23SrRNA gene mutation site of isolates resistant to Mp and drug resistance phenotype.   Methods　A total of 400 throat swab specimens of community-acquired respiratory tract infection were cultured to isolate Mp，the clinical isolates were identified by nested polymerase chain reaction，and the in vitro antibiotic sensitivity test was performed for identifying macrolide-resistant isolates through the minimal inhibitory concentration(MIC)．The sequences of macrolide-resistant 23SrRNA gene were detected. The sequences were compared to the corresponding sequences of M129． The relationship between mutation site and drug resistance phenotype was analyzed.  Results　A total of 50 Mp were isolated from 400 throat swab specimens．Of the 50 isolates，32 isolates were susceptible to macrolide，and 18 isolates were resistant to macrolide．The 18 clinical isolates appeared mutation A2063G，A2064G and A2067G，separately. A2063G showed 14 ring macrolide resistance．A2064G showed 14 and 16 ring macrolide resistances. A2067G showed josamycin resistance.  Conclusions　Mp to macrolide resistance is serious，and the mutation of 23SrRNA gene is a predominant mechanism that contributes to the macrolide resistance. Through the analysis of 23SrRNA gene mutation site and drug resistance phenotype，the clinical Mp drug resistance situation is obtained. The theoretical guidance for reasonable selection and application of antibiotics is provided.

Analysis on the correlation of CD40-1C/T gene polymorphism with children asthma

DU Jiangang 1，XU Hongxing 1，JIE Qiongfei 1，ZHONG Qiao 1，WU Yuanjian 1，XU Jicheng 2

2013, 28(1):  17-20. 

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Objective　To investigate the distribution of CD40 gene 5′untranslated region-1 site C/T gene polymorphism，and to study its relationship with the risk of asthma in local asthma children.   Methods　A total of 134 asthma children were enrolled，and 113 healthy children from healthy examination subjects were as control group in asthma and contol groups. Polymerase chain reaction-restriction fragment length polymorphism（PCR-RFLP）was used to determine the genotype and the allele frequencies of CD40-1C/T gene. The serum total level of IgE was determined by immunonephelometry. The relationship of CD40-1C/T gene polymorphism with the risk of asthma and IgE level was analyzed.  Results　The distributions of the C and T allele frequencies between asthma and control groups had no statistical significance （P＞0.05). The distribution difference of CC and CT+TT genotype frequencies was statistically significant （P＜0.05). The risk of CC genotype was 1.75 times higher than that of CT+TT genotype [95%confidence interval （CI）：1.01-3.04].There were significant differences in the level of IgE between CC genotype and CT or TT genotype in asthma group （P＜0.05).  Conclusions　CC genotype of CD40 gene 5′untranslated region -1 site might be associated with the development of asthma in local asthma children.

Diagnosis significance of serum copper biochemical examination in patients with hepato-lenticular degeneration

WU Junxia，YANG Qian，XUE Mingyue，LIU Qingyun，HU Jiyuan

2013, 28(1):  21-24.  DOI: 10.3969/j.issn.1673-8640.2013.01.005

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Objective　To investigate the levels of serum copper biochemical examination [copper ion（Cu²⁺），copper oxidase（SCO）and ceruloplasmin（CP）] in patients with hepato-lenticular degeneration（HLD）and their significance for the diagnosis of HLD.   Methods　By atomic absorption spectrophotometry，hydrochloric acid-p-phenylenediamine colorimetric method and immune turbidimetric method，288 patients with HLD and 90 healthy controls were measured for serum Cu²⁺，SCO and CP levels. The 285 patients with decreasing serum copper biochemical examination levels were classified according to clinical types （42 cases of pseudo-sclerosis type，49 cases of brain-visceral hybrid type，138 cases of liver type and 59 cases of Wilson type） and the age（under 10 years old group，11 to 20 years old group，21 to 30 years old group and ≥31 years old group），and the results of serum copper biochemical examination were compared. Results　In the 288 HLD patients，the results of serum copper biochemical examination of 3 cases were normal，and the abnormality rate was 98.96%. In 285 HLD patients with decreasing levels of serum copper biochemical examination，the serum level of Cu2+ was（2.46±1.73）μmol/L，the serum level of SCO was 0.027 （0.005-0.248）U/L，and the level of CP was （55.07±31.87）mg/L，which were significantly lower than those in controls [(15.09±2.42）μmol/L，0.492（0.269-0.798）U/L and（306.37±46.84)mg/L ]（P＜0.01）. The SCO level in Wilson type group was significantly lower than those in the liver type and pseudo-sclerosis type groups（P＜0.01），and the CP level was significantly lower than those in the liver type and brain-visceral hybrid type groups（P＜0.05，P＜0.01）. The serum Cu2+ level in pseudo-sclerosis type group was higher than those in the other clinical type groups（P＜0.05，P＜0.01）. The serum Cu2+ level was the lowest in under 10 years old group，and the results of serum copper biochemical examination in ≥31 years old group were higher than those in the other age groups. The areas under the receiver operating characteristic（ROC）curve（AUC）were 0.995，0.994 and 0.996，respectively. The optimal cut-off values were 6.34 μmol/L，0.121 U/L and 123.1 mg/L，respectively. The sensitivity，specificity and false identification rate were 95%，100% and 0%. Conclusions　Serum copper biochemical examination is the most convenient screening program for diagnosing HLD. For patients with HLD，the best diagnostic cut-off values are as follows：Cu2+ ≤ 6.34 μmol/L，SCO ≤0.121 U/L and CP ≤123.1 mg/L.

Clinical significance of myeloperoxidase in patients with chest pain

MA Qinghua 1，DENG Aiyun 1，ZHANG Zheng 1，LIU Xingrong 2，PAN Ming 1，SHEN Xiping 1，BAI Ming 1，WANG Zhilu 1

2013, 28(1):  25-29.  DOI: 10.3969/j.issn.1673-8640.2013.01.006

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Objective　To investigate the relationship between myeloperoxidase（MPO）concentration and the onset and progression of acute coronary syndrome（ACS）and the clinical significance of MPO in patients with chest pain.   Methods　A total of 78 patients with chest pain were enrolled and classified into 41 patients with ACS，17 patients with stable angina pectoris（SAP）and 20 controls according to the guideline of American College of Cardiology(ACC)/ American Heart Association（AHA). MPO concentration was measured by enzyme-linked immunosorbent assay. White blood cell（WBC)，neutrophil（Neu），red blood cell（RBC)，platelet（PLT)，total cholesterol（TC)，low-density lipoprotein cholesterol（LDL-C)，high-density lipoprotein cholesterol（HDL-C)，triglyceride（TG)，creatine kinase MB isoenzyme（CK-MB)，aspartate aminotransferase（AST)，fasting blood glucose（FBG)，lactate dehydrogenase（LDH）and high sensitive C reactive protein（hs-CRP）were detected. The correlations between MPO and other parameters were analyzed. By the receiver operating characteristic（ROC）curve，the area under ROC curve（AUC）was calculated. The diagnostic performance of MPO for ACS was evaluated. This technology was compared with the diagnostic methods of clinical manifestation analysis and coronary angiography based on the measurement of Kappa concordance. Results　The plasma MPO concentration in patients with ACS was significantly higher than those in patients with SAP and controls （P＜0.05），and the MPO concentration in patients with SAP was higher than that in controls （P＜0.05). The positive correlation was observed between MPO concentration and Neu，CK-MB and the severity of disease （r＝0.288，0.469 and 0.757，P＝0.018，0.043 and 0.000). MPO concentration did not correlate with age，hs-CRP，TC，LDL-C，HDL-C，TG，AST，FBG，LDH，RBC and PLT （P＞0.05). The cut-off value for MPO was identified as 212.59 μg/L（AUC＝0.927，P＝0.000）. In 41 patients with ACS，39 cases were MPO positive，and 2 cases were MPO negative. In 37 non-ACS patients（17 patients with SAP and 20 controls)，5 cases were MPO positive，and 32 cases were MPO negative. The sensitivity was 95.1%，the specificity was 86.5%，and the total concordance rate was 91.0%. The false negative rate（missed-diagnosis rate）was 4.9%，and the false positive rate（misdiagnosis rate）was 13.5%. The positive and negative predictive values were 88.6% and 94.1%. The Kappa value was 0.819（P＝0.000)，and the concordance of the 2 methods was good. Conclusions　MPO is a novel marker of inflammation for the diagnosis of ACS and has an important clinical significance for the identification of ACS.

The application of combined detection of the NT-proBNP and cTnI in the diagnosis and treatment of acute heart failure patients

ZHU Xiaopin，WANG Chenggang

2013, 28(1):  30-32.  DOI: 10.3969/j.issn.1673-8640.2013.01.007

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Objective　To investigate the application of combined detection of the N-terminal pro-B-type natriuretic peptide（NT-proBNP）and cardiac troponin I（cTnI）in the diagnosis and treatment of acute heart failure（HF）patients in emergency department.   Methods　Enzyme-linked immunosorbent fluorescence assay was used to determine plasma NT-proBNP and cTnI levels in 708 patients with cardiovascular diseases（195 patients with HF and 513 patients with non-HF). Acoording to the New York Heart Assoeiation（NYHA）cardiac function grades，the HF patients were chassified into Grade Ⅰ-Ⅳ. The results were expressed by medians（quartiles). The results were analyzed statistically. Results　The levels of NT-proBNP and cTnI in HF group were 3 718（1 035-12 258）ng/L and 7.18（2.05-14.16）μg/L，respectively，which were significantly higher than those in [145（30-585）ng/L for NT-proBNP and 0.66（0.06-1.99）μg/L for cTnI，P＜0.01]. Statistical significance was also observed when comparing NT-proBNP and cTnI levels of the patients with HF at NYHA Grade Ⅱ，Ⅲ and Ⅳ （P＜0.05). Furthermore，the sensitivity（99%）and specificity（90%）for the diagnosis of HF with the combined detection of NT-proBNP and cTnI were much higher than those with either the detection of NT-proBNP or cTnI alone（P＜0.05). Conclusions　The combined detection of NT-proBNP and cTnI has valuable significance in the diagnosis and treatment of HF in emergency department.

Changes of random urine total protein-to-creatinine ratio among chronic renal failure patients in 1 d

HU Yao 1，JIANG Guoying 2，LIU Ruilai 1，Lü Yuan 1，LIU Chunfang 1

2013, 28(1):  33-36.  DOI: 10.3969/j.issn.1673-8640.2013.01.008

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Objective　To research the changes of random urine total protein（TP)-to-creatinine（Cr）ratio in 1 d.   Methods　After being determined for 24 h urine protein，a total of 28 patients with renal inadequacy were enrolled. The endogenous creatinine clearance rate（Ccr）was ＞10mL/min. When collecting 24 h urine，about 1.5mL urine was taken out at different point as random urine specimen，and the others were put into the 24 h urine specimen quantitation collections. The values of TP，Cr and TP/Cr ratio in random and 24 h urine specimens were detected and calculated. The 24 h urine specimens with high，middle and low protein concentrations were used to analyze laboratory error [between-run coefficient of variation（CV）]. Results　The CV of TP and TP/Cr ratio with high，middle and low protein concentrations were ＜ 5% in the clinical acceptable range. In 11 of 28（40%）patients，the differentiation between the random urine TP/Cr ratio and 24 h urine TP/Cr ratio wasn′t significant in statistics （P＞0.05）. The 24 h urine protein can be calculated by random urine TP/Cr ratio. After calculating the maximum clinical acceptable range，there were 17（60%）patients whose TP/Cr ratio having a significant differentiation between the random urine TP/Cr ratio and 24 h urine TP/Cr ratio in statistics （P＜0.05）. The patients whose TP/Cr ratio of random urine specimens in 1 d had a highly significant differentiation，evenly reached 30 folds. It was hard to get right amount of 24 h urine protein through predicting random urine TP/Cr ratio. The determination of 24 h urine protein can not be replaced by random urine TP/Cr ratio. To many patients，the urine protein and TP/Cr ratio in random urine specimens showed negative results in some points，although the results of 24 h urine protein were abnormal obviously. Conclusions　These results show that random urine TP/Cr ratio in 1 d has significant changes for many patients. Therefore，24 h TP should be used as an alternative to analyze renal protein.

Expression and significance of decoy receptor 3 in HBeAg negative chronic hepatitis B patients

HOU Yanqiang 1，LIANG Dongyu 1，PENG Liang 1，LOU Xiaoli 1，LIU Tao 1，ZHANG Lurong 2

2013, 28(1):  37-39.  DOI: 10.3969/j.issn.1673-8640.2013.01.009

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Objective　To investigate the expression and significance of decoy receptor（DcR3）in hepatitis B virus（HBV）e-antigen（HBeAg）negative chronic hepatitis B（CHB）patients.   Methods　Serum DcR3 levels were measured by enzyme-linked immunosorbent assay（ELISA）in 80 HBeAg negative CHB patients and 96 healthy subjects. Alanine aminotransferase（ALT）was determined by automatic analyzer. Serum HBV DNA was determined by polymerase chain reaction（PCR).  Results　DcR3 level in HBeAg negative CHB patients [1.58(0.48-3.23)ng/mL]was significantly higher than that in healthy controls [0.83(0.16-2.32）ng/mL，P＜0.000 1]. DcR3 level was correlated positively with HBV DNA （r＝0.704，P＜0.000 1）and ALT （r＝0.732，P＜0.000 1).  Conclusions　DcR3 has high expression and correlates with HBV DNA and ALT in HBeAg negative CHB patients，and DcR3 may serve as a diagnosis and treatment monitoring indicator for HBeAg negative CHB patients.

The diagnosis significance of serum procalcitonin in children with community-acquired pneumonia

LU Junyan 1，JIANG Zhigang 1，ZHOU Weixin 1，JIANG Zhijun 1，XIE Qingyun 2，YANG Zeyan 2

2013, 28(1):  40-43.  DOI: 10.3969/j.issn.1673-8640.2013.01.010

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Objective　To investigate the differential diagnosis significance of serum procalcitonin（PCT）in children with bacterial and viral infected community-acquired pneumonia（CAP)，and compare the accuracies and application of C reactive protein（CRP)，erythrocyte sedimentation rate（ESR）and white blood cell（WBC）count.   Methods　A total of 79 patients with CAP（20 cases with lobar pneumonia and 59 cases with bronchopneumonia）and 21 healthy controls were enrolled. PCT，CRP，ESR and WBC count were determined. Receiver operating characteristic（ROC）curve analysis was performed. The 79 patients with CAP were classified into ＜2 years old group（31 patients)，2-5 years old group（18 patients）and ＞5 years old group（30 patients）according to their ages，and their PCT，CRP，ESR and WBC counts were compared. Results　Serurn PCT，CRP，ESR and WBC count were significanthy higher in CAP group than in control group（P＝0.000，P＜0.01). Serum PCT，CRP，ESR and WBC count in lobar oneumonia group were higher than those in bronchopneumonia group（P＜0.01). The level of PCT in ＞5 years old group were higher than that in ＜2 years old group（P＝0.000)，and the levels of CRP，ESR and WBC count had no statistical significance among the different age groups（P＞0.05). The area under receiver sperating chara cteristic（ROC）curve of PCT was 0.97 [95% confidence inter val （CI）：0.93-1.00]，which was higher than those of CRP [0.91（95%CI：0.78-0.99)] and ESR [0.87（95% CI:0.78-0.96)]（P＜0.01). The area of WBC count [0.95（0.95%CI：0.91-0.98)] had no statistical significance（P＝0.14). The best cut-off value of PCT was 1 ng/mL，and the sensitivity and specificity were 94.1% and 82.3%. Conclusions　Serum PCT level is a better marker than CRP and ESR for the diagnosis of CAP in children，and can be used as a reliable indicator of the early diagnosis for clinicians and rational use of antibiotics.

Chromosome karyotype determination analysis in 850 genetic counseling subjects

HAN Jing

2013, 28(1):  44-46.  DOI: 10.3969/j.issn.1673-8640.2013.01.011

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Objective　To investigate peripheral blood chromosome determination and chromosome karyotype analysis among high risk population.   Methods　A total of 850 high risk genetic counseling subjects were enrolled in the study，who underwent chromosome determination and chromosome karyotype analysis. Results　Among the 850 cases，there were 258 cases（30.35%）with chromosomal abnormalities. Of them，145 cases（56.20%）were mental retardation，86 cases（33.33%）were reproductive disorders，and 27 cases（7.75%） had a family history. Conclusions　The incidence rate of chromosomal abnormalities is quite high in high risk population，so it is very important for chromosome determination among high risk population，which can find the cause，improve the diagnosis and treatment level，improve population quality for eugenics，and reduce and avoid the occurrence of chromosomal diseases.

The variation of immature platelet fraction in patients with thrombocytopenic diseases

JIANG Weiyan，JIANG Minghua，WU Yizhong，ZHANG Zhaohua，CHEN Xiaojian

2013, 28(1):  47-50.  DOI: 10.3969/j.issn.1673-8640.2013.01.012

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,Objective　To investigate the variation of immature platelet fraction（IPF)，high-fluorescence intensity of immature platelet fraction（H-IPF)，absolute value of immature platelet（IPF#）and mean side fluorescence intensity of platelet(PLT-X）in patients with thrombocytopenic diseases and their clinical significance in the thrombocytopenic diseases.   Methods　The platelet（PLT)，IPF，H-IPF，IPF# and PLT-X of peripheral blood in 86 patients with thrombocytopenic diseases [50 cases of idiopathic thrombocytopenic purpura（ITP)，15 cases of aplastic anemia（AA）and 21 cases of acute leukemia（AL)]，32 patients with myeloproliferative disorders（MPD）[11 cases of chronic myelogenous leukemia（CML)，16 cases of essential thrombocythemia(ET）and 5 cases of polycythemia vera（PV)] and 50 healthy subjects were determined by automatic hematology SYSMEX XE-5000 analyzer. According to the results of PLT，the 50 cases of ITP were classified into ≤30×10⁹/L，(＞30-＜60)×10⁹/L and（60-＜100)×10⁹/L groups，and the results for IPF were compared.  Results　The IPF，H-IPF，IPF# and PLT-X in ITP group were（19.8±12.7）%，6.7（4.7-12.3）%，3.1（2.0-12.1）×10⁹/L and（27.8±8.6）ch，respectively. The IPF，H-IPF，IPF# and PLT-X in AA group were（5.6±2.5）%，1.9（0.7～4.0）%，0.8（0.4～1.4）×10⁹/L and（17.3±2.4）ch，respectively. The IPF，H-IPF，IPF# and PLT-X in AL group were（6.3±3.4）%，2.1（1.2～4.1）%，2.4（1.5～3.2）×10⁹/L and（18.7±3.0)ch，respectively. The IPF，H-IPF，IPF# and PLT-X in MPD group were（3.1±1.6）%，0.9（0.7～1.4）%，19.2（14.0～22.5）×10⁹/L and（16.9±2.3)ch，respectively. The IPF，H-IPF，IPF# and PLT-X in healthy subject group were（4.1±1.3）%，1.2（1.0～1.7）%，9.3（7.4～12.1）×10⁹/L and（18.4±1.5）ch，respectively. Compared with the AA，AL，MPD groups and healthy subject group，the IPF，H-IPF and PLT-X of ITP group wrer higher（P＜0.05)，and the difference in the IPF# between ITP group and the other 3 groups was significant（P＜0.05)，but there was no statistical significance in the IPF# between ITP group and healthy subject group（P＞0.05). There was no statistical significance for IPF in the different groups of ITP（P＞0.05）.

Expression of survivin and CD34 in bone marrow cells of patients with acute myeloid leukemia

GU Dawei，MA Li

2013, 28(1):  51-56.  DOI: 10.3969/j.issn.1673-8640.2013.01.013

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Objective　To investigate the relationship of bone marrow cell expression of survivin and cluster of differentiation 34（CD34）in the development of acute myeloid leukemia（AML).  Methods　The survivin and CD34 were detected by flow cytometry in 62 patients with AML and 31 patients with non-malignant hematopathy. Results　The percentage of the expression of survivin^＋/CD34^＋ in bone marrow cells of patients with AML [(13.74±10.92)%] was obviously higher than that of control group [(2.53±1.85)%，P＜0.01]，and that of remission group [(4.01±1.41）%] was lower than that of non-remission group[(23.83±20.42)%，P＜0.01]. The percentage of the expression of survivin^＋/CD34- in bone marrow cells of patients with AML [(22.07±16.95)%] was obviously higher than that of control group [(12.29±8.80)%，P＜0.05]，and that of remission group [(14.32±7.53)%] was lower than that of non-remission group [(29.83±26.37)%，P＜0.05]. Conclusions　In the detection of hematopathy，the expression of survivin in bone marrow cells has the role of outcome with screening. The expression of survivin has closer relationship with the development of leukemia than CD34，and has a certain significance in the treatment and prognosis assessment of AML.

Application of urine protein electrophoresis in early diabetic nephropathy

RAO Hui 1，ZANG Li 2，RUN Yuanmin 1，PI Youjun 1

2013, 28(1):  57-60.  DOI: 10.3969/j.issn.1673-8640.2013.01.014

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Objective　To investigate the application significance of urine protein electrophoresis in early diabetic nephropathy（DN).   Methods　Urine protein electrophoresis was used to analyze 48 type 2 diabetes mellitus patients with urine microalbumin（mAlb）15-300 mg/24 h. Dry chemistry urine protein，urine glucose，urine occult blood and glycosylated hemoglobin were also measured. A total of 10 healthy controls were enrolled and analyzed by urine protein electrophoresis. Results　The 8（17%）patients showed tubular proteinuria，36（75%）patients showed glomerular proteinuria[32（67%）cases：microalbuminuria and 4（8%）cases：selective glomerular proteinuria]，and 4（8%）patients showed no urine protein band by urine protein electrophoresis. There were statistical significances in the scan percentage of urine protein among the all groups（P＜0.05）. Among the 10 healthy controls，no urine protein band was detected. The 2 of 8 cases of tubular proteinuria showed urine mAlb ≤30 mg/24 h. There was a positive correlation between the percentage of tubular urine proteins and glycosylated hemoglobin（r＝0.300）. Conclusions　Tubular dysfunction accounts for a considerable proportion in early DN. Urine protein electrophoresis combined with urine mAlb determination can decrease the probability of missed diagnosis of tubular proteinuria in early DN.

Detection of interleukin-22/17-producing lymphocytes subpopulations in peripheral blood of human by flow cytometry and application in patients with tuberculosis

LIANG Hua 1，LIANG Youbao 2，HE Wenxin 3，YANG Junping 1，SUN Guohong 1，YANG Xiaojun 1，LI Baiqing 3

2013, 28(1):  61-66.  DOI: 10.3969/j.issn.1673-8640.2013.01.015

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Objective　To establish a method of detecting different interleukin 22（IL-22）producing lymphocyte subpopulations in human peripheral blood by flow cytometry（FCM)，and investigate its application in patients with tuberculosis（TB）and realize the role of IL-22 producing lymphocyte subpopulations in anti-infection immunity against TB.   Methods　Peripheral blood samples collected from 22 TB patients and 18 healthy controls were mixed with RPMI-1640 culture medium at the scale of the same volume. After adding phorbol ester（PMA）and calcium ionomycin mixed for 2 h culturing on the conditions of 37℃ and 5% CO₂，monensin was added and continued being cultured for 4 h. The cells were collected and stained surface molecular or intracellular with fluorochrome-conjugated monoclonal antibodies. The respective proportions of different IL-22 and IL-17 producing lymphocyte subpopulations were detected by FCM. Results　The condition that the concentration of PMA was 100ng/mL and that of ionomycin was 1μg/mL with 2 h being stimulated and cultured could produce IL-22 effectively. The proportions of IL-22 producing gamma delta T cells in TB patients（7.37%）were obviously higher than those in healthy controls（1.81%，P＜0.05). No statistical significance was found between the proportions of IL-22 producing positive cells in CD4^＋T，CD8^＋T，natural killer（NK）cells and B cells in TB patients and those in healthy controls （P＞0.05). No statistical significance was found between the proportions of Th22 and Th17 in TB patients and those in healthy controls （P＞0.05). Conclusions　The established easy，complete and reliable method for the detection of IL-22 producing lymphocyte subpopulations and IL-22 producing gamma delta T cells could act immunological effects in anti-infection immunity against TB.

Mast cells exerting protumorigenic role in the early phase of gastric carcinoma and its biological significance

LIU Yanan，LIN Lihui，WANG Juan，LI Jia，PENG Xia，XIAO Hui，LI Fei，DAI Huirong，LI Li

2013, 28(1):  67-71.  DOI: 10.3969/j.issn.1673-8640.2013.01.016

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Objective　To investigate the correlation of mast cell with tumorigenesis and survival time through analyzing the mast cell counts in the tissues of gastric carcinoma，precancerous lesions of gastric carcinoma and benign gastric lesions.  Methods　The pathological and follow-up data of 59 patients with gastric carcinoma，40 patients with precancerous lesions of gastric carcinoma and 39 patients with benign gastric lesions were analyzed retrospectively. The mast cell specific tryptase was labeled by immunohistochemistry. The counts and distribution of mast cells in the tissues of gastric carcinoma（high，middle and low differentiation）were analyzed. The expression in the tissues of gastric carcinoma was compared with those in the tissues of precancerous lesions of gastric carcinoma and benign gastric lesions，and the correlation of mast cell with pathological data and survival time was analyzed simultaneously. Results　The mast cell counts were higher in the tissues of gastric carcinoma and precancerous lesions of gastric carcinoma than those of benign gastric lesions （P＜0.05 and P＜0.05），and the mast cell counts in the tissues of precanerous lesions of gastric carcinoma were higher than those of gastric carcinoma （P＜0.05). According to the medians of mast cell counts（28.43），the results between gastric carcinoma group and precancerous lesions of gastric carcinoma group were compared，and the survival times of high-levle and low-level mast cell counts in gastric carcinoma and precancerous lesions of gastric carcinoma groups had no statistical significance （P＞0.05 and P＞0.05). There were correlations of mast cell counts with Helicobacter pylori and Ki-67 expression （P＜0.05 and P＜0.05). Conclusions　The varied mast cell counts show that mast cells correlate the protumorigenesis，especially Helicobacter pylori induced gastric carcinoma. The correlation of mast cell and Ki-67 shows mast cell exert tumor proliferation and progression，but there is no correlation with the degree of differentiation. There is no obvious correlation of mast cell with survival time.