Table of Content

05 July 2012, Volume 27 Issue 7

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Comparison investigation on the results of different detection system blood coagulation analyzers

WANG Lejian;SHI Chunjuan;LI Fei

2012, 27(7):  527-530. 

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Objective　To investigate the comparability of different detection system blood coagulation analyzers，in order to achieve the efficiency of quality control. Methods　With STA compact and CA-1500 blood coagulation analyzers，the quality control material and 100 comparison blood samples（concentration covered biology reference interval） were determined. The measured results were analyzed and compared according to American Clinical Laboratory Improvement Amendment 88（CLIA′88） allowable total error 1/2 comparison and correlation（r＞0.975） as judgment standard. The instrument comparability and correlation were observed. Results　When STA compact and CA-1500 blood coagulation analyzers measuring samples without icterus and lipaemia，the results of international normalized ratio（INR） and low-value fibrinogen（Fbg） were consistent. The relative deviation was within allowance range，and the correlation was good（r＞0.975）. The result of activated patial thromboplastin time（APTT） correlation was fair，but the relative deviation was not allowed. The results of INR，APTT and Fbg was inconsistent when the samples was icterus or（and） lipaemia，and the relative deviation was not allowed and the correlation was bad（r＜0.975）. Conclusions　The comparability，correlation and differences can be reflected by comparing the results of different detection system blood coagulation analyzers

The performance evaluation of COBIO automatic urine system

WEI Yunmei;YI Guangming;XIONG Jun

2012, 27(7):  531-533. 

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Objective　To evaluate the performance of COBIO automatic urine system（combi XL analyzer and COBIO xs automatic urine sediment analyzer）. Methods　The repeatability，linearity，accuracy and sensitivity were determined by COBIO automatic urine system. Results　The within-run coefficients of variation（CV%） of white blood cell in high，middle and low-value samples were 4.5，6.0 and 10 respectively. They were 2.4，5.0 and 13.8 for red blood cell，and 4.8，7.0 and 13.5 for epithelial cell. The correlation coefficient of red blood cell was 0.999 with 5-1210/μL. The correlation coefficient of white blood cell was 0.998 with 4-498/μL. The accuracies of red blood cell，white blood cell，epithelial cell，cast cell，calcium oxalate，uric acid crystallization and Candida were 99%，98%，93%，58%，93%，95% and 90%，recpectively. Conclusions　COBIO automatic urine system has the advantages of automation，high testing speed，accuracy and low cost，and it has a good significance in clinical application.

Establishment of reference intervals of the white blood cell VCS parameters and their application in sepsis and leukemia

WANG Hai;REN Feiyao;PENG Wenhong;LAN Yating;WANG Jianbiao

2012, 27(7):  534-539. 

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Objective　To establish the reference intervals of the white blood cell volume，nucleus-cytoplasm ratio and the granules of plasma in healthy subjects，and to evaluate the clinical application in screening sepsis and leukemia. Methods　Peripheral blood samples from 290 healthy subjects（including 137 males and 153 females），42 infective patients（including 23 males and 19 females） with positive blood culture（excluding any hematological disease）and 260 patients（including 147 males and 113 females） with various leukemia were collected. Then，the reference intervals of the white blood cell VCS parameters were established and analyzed statistically. The cut-off values with high sensitivity and specificity were chosen by receiver operating characteristic（ROC） curves in screening sepsis and leukemia. Results　The reference intervals of VCS parameters were obtained by the data analysis. Mean channel of lymphocyte conductivity（MLC） was a useful indicator in discriminating the healthy subjects and patients with sepsis and leukemia. The parameters of mean channel of neutrophil conductivity（MNC） could be used as indicators in distinguishing sepsis from leukemia. Myeloid leukemia and lymphocytic leukemia could be discriminated by mean channel of lymphocyte volume distribution width（MLV-SD）. Mean channel of nononuclear cell conductivity distribution width（MMC-SD） was a good indicator in distinguishing acute myelogenous leukemia（AML） from chronic myelogenous leukemia（CML）. mean channel of lymphocyte volume（MLV） was useful in discriminating acute lymphoblastic leukemia（ALL） and chronic lymphoblastic leukemia（CLL）. Conclusions　The establishment of reference intervals of the white blood cell VCS parameters makes it possible to deciding earlier whether there is an infection with bacteria and leukemia. VCS parameters not only reflect morphological changes of immature cells in peripheral blood samples when it comes to sepsis or leukemia，but also is used to screen leukemia quickly，economically and effectively.

Analysis of the pathogens isolated from blood specimens of neonates and their antibiotic resistance

SONG Haiying;CHENG Xiaojing;ZHANG Wenrong

2012, 27(7):  540-543. 

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Objective　To investigate the distribution and antibiotic resistance of pathogens isolated from blood specimens of neonates. Methods　Biphasic blood culture bottle was used to culture the blood of the neonates. Isolated strains were identified，and their drug sensitivity tests were completed by VITEK 32 system．Results　The results showed that Gram-positive cocci accounted for 70.6%，in which Staphylococcus epidermidis accounted mainly，and Gram-negative bacteria accounted for 28.4%． Gram-positive cocci were sensitive to vancomycin，and Gram-negative bacteria were sensitive to carbapenem. Conclusions　The antibiotic resistance determination of pathogens isolated form blood specimens of neonates should be paid attention. Choosing rational antibiotics can reduce the antibiotic resistance and improve the clinical curative effect.

Accuracy evaluation of different antibiotic susceptibility testing for mucoid isolates of Pseudomonas aeruginosa

LAN Kai;ZHANG Weizheng;LUO Qiang;ZHOU Qiang;ZHANG Wen;TANG Xiaolong

2012, 27(7):  544-548. 

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Objective　To investigate the accuracy of different methods of antibiotic susceptibility testing for mucoid isolates of Pseudomonas aeruginosa（PA）. Methods　A total of 66 isolates of mucoid PA were detected by Kirby-Bauer（K-B） method，ATB PSE5 kits，MicroScan WalkAway 96 plus，VITEK 2 and micro-broth dilution method. The micro-broth dilution method was adopted as reference method，and the other methods were as test methods. Results　Compared to micro-broth dilution method，the complete accordances（CA） of K-B method，ATB PSE5 kits and MicroScan were 92.63%，88.89% and 89.38%. The very major errors（VME） of the 3 methods were 0.30%，0.83% and 0.34%，respectively. The major errors（ME） of the 3 methods were 3.13%，5.83% and 5.19%. The minor errors（MIE） of the 3 methods were 3.94%，4.44% and 5.07%. The results lack in VITEK 2，since the relevant data were not analyzed. Conclusions　K-B method is a reliable method for mucoid PA antibiotic susceptibility testing，especially prolonging the incubation time to 48 h or incubating in CO2 incubator for 24 h.

Research on the correlationship of rs7903146 polymorphism in the TCF7L2 gene with type 2 diabetes mellitus

LIU Guozheng;YAN Wenqiang;LIU Shuang;ZHANG Zhenjian;ZHOU Zhongcheng;JIANG Hao

2012, 27(7):  549-553. 

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Objective　To investigate the correlationship of transcription factor 7-like 2（TCF7L2） gene rs7903146 polymorphism and Chinese Han population in Suizhou district with type 2 diabetes mellitus. Methods　A total of 458 patients with type 2 diabetes mellitus（232 males and 226 females） had been diagnosed，and 186 healthy subjects（91 males and 95 females） with fasting blood glucose ＜5.6 mmol/L and without other systemic diseases were enrolled. The clinical indicators and glucose metabolism and lipid metabolism related indicators were measured. The polymerase chain reaction（PCR） TaqManTM fluorescence probe technique was used for allelic typing. The TCF7L2 gene rs7903146 polymorphism was analyzed. Results　TCF7L2 gene rs7903146 polymorphism was consistent with Hardy-Weinberg equilibrium law，and the frequencies of TCF7L2 gene rs7903146 C/C，C/T and T/T genotypes in type 2 diabetes mellitus group were 92.2%，7.6% and 0.2%，and the frequencies in the control group were 92.5%，7.5% and 0. No statistical difference between the 2 groups was found by χ² test. T allele frequencies in the type 2 diabetes mellitus group and control group were 4.0% and 3.8%，without statistical significance. Conclusions　TCF7L2 gene rs7903146 polymorphism is not the main pathogenic gene among Chinese Han population in Suizhou district with type 2 diabetes mellitus.

RASSF1A gene methylation and its clinical significance in leukemia patients

CHEN Weiqun;HUANG Chen;ZHAN Xiyan;LIU Shuiyi;YU Jie;ZHANG Hongmei;LU Zhongxin

2012, 27(7):  554-556. 

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Objective　To research Ras association domain family 1A（RASSF1A） gene promoter region methylation in bone marrow of leukemia patients and its clinical significance. Methods　Methylation of RASSF1A gene promoter region was detected by methylation specific polymerase chain reaction（MS-PCR） in the bone marrow of 86 leukemia patients，and the difference of different kinds of leukemia was analyzed. A total of 60 non-leukemia subjects were enrolled as controls. Results　Twelve of 86（13.95%） leukemia patients had RASSF1A gene promoter region methylation. No methylation was found in bone marrow of the control group，and there was a statistical significance（P＜0.05）. RASSF1A gene methylation ratio in lymphoid leukemia（23.1%） was higher than that in myeloid leukemia（6.4%）（P＜0.05）. Conclusions　Leukemia patients have RASSF1A gene methylation. The ratio of RASSF1A gene methyaltion in lymphoid leukemia is obviously higher than that in myeloid leukemia. RASSF1A gene methyaltion may be a significant parameter for diagnosis and genotyping of patients with leukemia.

Comparison on detection of YMDD mutation in hepatitis B virus by three methods

LIU Wenju;JIANG Peixue;ZHU Ruxiang

2012, 27(7):  557-560. 

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Objective　To analyze comparatively and find a sensitive detection method for hepatitis B virus（HBV） YMDD mutation. Methods　A total of 166 samples from patients with HBV infection were collected，including 116 cases treated by lamivudine for 1 year and 50 cases treated by telbivudine for 1 year. The comparison of 3 HBV YMDD mutation detection methods real-time polymerase chain reaction（PCR），high temperature ligase detection reaction（LDR） and sequencing was analyzed. In addition，the sensitivities of the 3 methods were detected. Results　The positive rates of LDR，real-time PCR and sequencing were 23.49%，21.68% and 19.27%. The specificities of the 3 methods were 100%. In the cases treated by lamivudine，there were 38 cases with mutation（12 YIDD and 26 YVDD）. There was only 1 case with YIDD mutation in the cases treated by telbivudine. Conclusions　The LDR for detecting YMDD mutation is suitable in clinical application，and YVDD mutation is more common than YIDD mutation.

Evaluation on the methodology of blood manganese determination by inductively coupled plasma atomic emission spectrometry

YU Fei;LI Meng;CAO Bo

2012, 27(7):  561-563. 

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Objective　To evaluate the methodology of determining blood manganese by inductively coupled plasma atomic emission spectrometry（ICP-AES），and apply the method in daily detection work. Methods　The whole blood samples were treated with nitric acid，and the concentration of the nitric acid was 1 mol/L. The supernatant fluid was collected. The concentrations of the whole blood manganese were determined by ICP-AES under the condition of 279.544 nm in 1 027 healthy children. The precision，recovery rate，sensitivity and anti-interference ability were evaluated，and the normal reference range was definited. Results　The within-run coefficient of variation（CV） was 6.2%-6.6%，the between-run CV was 7.1%-7.6%，and the total CV was 7.2%-7.8%. The relative standard deviation（RSD） was 1.66%-2.36%. The standardized recovery rate was 95.0%-96.9%. The linear equation was Y＝7 035.6X+759.02，and the correlation coefficient of the whole blood manganese was r＝0.999 7. The interference elements，K，Na，Cl，Ca and Mg（1∶〖KG-*2/5〗1，25 mg/L） and Fe（2 mg/L） were added into the blank solution and the standard reserve liquid of 100 μg/L manganese. The concentration of manganese had no obvious change. The normal reference range of whole blood manganese in 1 027 healthy children was（4.89±2.67） μg/L. Conclusions　ICP-AES for determining whole blood manganese has no obvious system error. This method is accurate and reliable，and can be applied as a conventional way to detect whole blood manganese.

Investigation on the interference of bilirubin and hemoglobin for different urinary albumin measurements

LIU Rui;LI Gang;CUI Xiaofan

2012, 27(7):  564-567. 

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Objective　To investigate the interference of bilirubin（Bil） and hemoglobin（Hb） in urine on different urinary albumin measurements. Methods　Bil and Hb，as interferents，performed interference test by 3 different urinary albumin measurements：immuno-nephelometry，immuno-turbidimetry and immuno-colloidal gold method，respectively. Results　Bil and Hb with different concentrations could affect the results of urinary albumin by all the 3 measurements，and the interferents rate increased with the increasing concentration of interferents. Hb mainly generated negative interference by the 3 methods，while the interferences of Bil by immuno-nephelometry and immuno-turbidimetry were positive，and by immuno-colloidal gold method was negative. In the 3 methods，when urine occult blood was "1+"，the absolute value of interference rate of Hb was 6.72%-17.43%. When urine Bil was "1+"，the absolute value of interference rate was 7.89%-22.94%.Conclusions　Bil and Hb could generate interference on urinary albumin measurements. When detecting urinary albumin from the patients with suspected high concentrations of Bil and Hb in urine，the interference factors should be considered.

Performance of urine podocalyxin and microalbuminuria in the diagnosis of diabetic nephropathy

YE Jun;LUO Hong;LI Kun;MA Wei

2012, 27(7):  568-570. 

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Objective　To study the significance of podocalyxin（PCX），microalbuminuria（mAlb） and the combined detection of urine PCX and mAlb in the early diagnosis of diabetic nephropathy（DN） by receiver operating characteristic（ROC） curve.  Methods　The detection results of PCX，mAlb，PCX+mAlb from 35 DN patients and 47 diabetes mellitus（DM） patients were evaluated by ROC curve. Results　PCX and mAlb of DN group were significantly higher than those of DM group（P＜0.01）.The areas under ROC curve of PCX，mAlb and PCX+mAlb in DN patients were 0.872，0.921 and 0.973 respectively. The single discrimination equation was predictive probability value（P）＝1/[1+e^{－（－5.030+0.067mAlb+0.097PCX）}]. When P≥0.565，it was positive and diagnosed as DN. Conclusions　By the evaluation of urine ROC curve，the combined detection of urine PCX and mAlb is better than PCX or mAlb individual detection for the diagnosis of DN.

Evaluation on the homocysteine detection by enzyme transition method and the early diagnostic significance of homocysteine to coronary heart disease

XING Xiaoguang

2012, 27(7):  571-574. 

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Objective　To investigate the significance of detecting homocysteine（Hcy） by enzyme transition method and the early diagnostic significance of Hcy to coronary heart disease. Methods　The enzyme transition method for detecting Hcy was evaluated by methodology for precision，linearity，interference，recovery and sensitivity tests，and was compared to high performance liquid chromatography（HPLC） method. The levels of Hcy among 60 coronary heart disease patients，55 healthy controls and 54 patient controls with normal blood lipid and glucose（Glu） levels and atherosclerosis family medical history were detected by enzyme transition method，and the levels of Glu，total cholesterol（TC） and triglyceride（TG） were detected. Results　The within-run imprecisions（CV） were 1.98% and 2.34% respectively. The between-run CV were 2.13% and 1.87% respectively. The interday CV were 2.47% and 1.33% respectively. The total CV were 3.85% and 2.41% respectively. There was a good linear relationship between 2-80 μmol/L（r²＝0.987 4）. Almost no significant interference can be observed in the enzyme transition method to detect Hcy of TG＜20 mmol/L，total bilirubin（TBil） ＜300 μmol/L and hemoglobin（Hb）＜10 g/L（P＞0.05）. The recovery rate was 98.9%-102.4%. The sensitivity was 0.27μmol/L. There was a good correlation between the enzyme transition method and HPLC method by linear-correlated analysis（r＝0.935 2），and there was no significant difference between the 2 methods（P＞0.05）. The positive correlation of Hcy with Glu and TG（r＝0.447 and 0.426，P＜0.01），and no correlation between Hcy and TC were shown（r＝0.146，P＞0.05）. The levels of Hcy in coronary heart disease group and patient control group were higher than those in healthy control group（P＜0.01）. The levels of TG and Glu in coronary heart disease group were higher than those in patient control group and healthy control group（P＜0.01）. Conclusions　Enzyme transition method is suitable for clinical laboratory with the advantages of good precision，high sensitivity，wide linear range，good anti-interference ability and accurate results. It has a significance for the early diagnosis of coronary heart disease when the level of Hcy rises.

Primary comparison of three automatic analyzers with same principle for glycosylated hemoglobin measurement

YAN Ying;ZHANG Chuanbao;ZHANG Jiangtao;MA Rong;WANG Donghuan;ZHANG Tianjiao;ZENG Jie;ZHOU Weiyan;CHEN Wenxiang

2012, 27(7):  575-578. 

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Objective　To compare TOSOH HLC-723 G7（HLC-723 G7），Bio-Rad D-10（D-10） and Arkray ADAMSTMA1c HA-8160（HA-8160） automatic glycosylated hemoglobin（HbA1c）analyzers with same principle ion-exchange high performance liquid chromatography for the measurement of HbA1c，and evaluate the difference among the results of the 3 automatic analyzers. Methods　The 58 patients′ samples were measured for 13 d by the 3 automatic analyzers respectively，when the low-and high-level HbA1c quality control samples were qualified. The homogeneity of variance and the one-way analysis of variance were tested with the quality control samples and patients′ samples，and the results were used to investigate the differences among the 3 analyzers. The scatter plot and difference plot of the 3 analyzers were made to check the linearity and difference. The data were used for linear regression equation，and the relative difference between each 2 analyzers was evaluated. Results　There was statistical significance among the results of the 3 analyzers for low-level quality control samples（P＜0.05），and there was no statistical significance for high-level quality control samples（P＞0.05）. The relativities were good among the 3 analyzers for measuring patients′ samples（P＞0.05）. When HbA1c level fell in the range of 4.9%-11.4%，the linear equation of HA-8160 and D-10 was Y＝0.963X + 0.179，r＝0.988，r²＝0.976，and the relative difference was －0.37%- 0.72%. The linear equation of HLC-723 G7 and D-10 was Y＝0.968X+0.523，r＝0.991，r²＝0.982，and the relative difference was －0.1%- 0.8%. The linear equation of HLC-723 G7 and HA-8160 was Y＝ 0.987X + 0.480，r＝0.984，r²＝0.969，and the relative difference was －0.13%- 1.12 %. Conclusions　At usual physiological and pathological ranges，there is no significant difference among the results measured by HLC-723 G7，D-10 and HA-8160 automatic analyzers.

Influence of alpha-lipoicacid on serum homocysteine in obese rats

YAN Guochao;SHAO Yun;QIN Xiaoxia;WU Yan

2012, 27(7):  579-584. 

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Objective　To observe the influence of alpha-lipoicacid（ALA） on serum homocysteine（Hcy） level in obese rats fed with high-fat diet（HFD）. Methods　A total of 10 rats were selected randomly from 50 Wistar rats，and were fed as a normal forage group，whereas the other 40 rats received a HFD for 10 weeks. Its obese rat model was evaluated，and obese rats were randomly classified into 2 groups：HFD group and HFD+ALA group who were injected with ALA for 2 weeks intraperitoneally at a dose of 30 mg/kg. The Hcy，fasting blood glucose（FBG），fasting insulin（FIns），triglyceride（TG），total cholesterol（TC），high density lipoprotein cholesterol（HDL-C），low density lipoprotein cholesterol（LDL-C） and non-esterified fatty acid（FFA）in serum were determined. The insulin resistance index（IRI） was calculated homeostatic model assessment of insulin resistance（HOMA-IR）.Results　The obese rat models of 32 obese rats were obtained after 10 weeks′ HFD. Compered to the normal forage group，HFD group gained a higher levels of Hcy，FIns，IRI，TC，TG，HDL-C and LDL-C（P＜0.05）. FBG and FFA had no obvious difference between the 2 groups（P＞0.05）. Compared to the HFD group，HFD+ALA group had a lower levels of Hcy，FIns，IRI，TC，TG，HDL-C，LDL-C and FFA，and FBG had no obvious difference. FFA levels in HFD+ALA group were significantly lower than those in the HFD group and the normal forage group（P＜0.05）. Conclusions　Obese rats induced by HFD have a high level of Hcy. ALA can improve insulin resistance，and reduce the serum Hcy level.

Establishment of the complement-mediated indirect immunofluorescence method for detecting anti-perinuclear factor and its primary clinical application

LI Yuqin;LI Lihe

2012, 27(7):  584-587. 

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Objective　To establish a complement-meditated indirect immunofluorescence method（two-step method） for detecting anti-perinuclear factor（APF） and apply it primarily. Methods　The epithelial cells of oral cheek from non-smoking healthy and non-connective tissue disease subjects were used as substrate. The diluted serum，complement and fluorescent label anticomplement C_3C were added. The nuclear fast red-glycerol was performed. By microscopy，the sizes with round or oval homogeneous fluorescent granules in cytoplasm or around the nucleus were as positive. The positive cells accounted for 50% of total cells were regarded as APF-positive，and the results were compared before and after inactivating serum complement. The APF of 76 rheumatoid arthritis（RA） patients at activity and remission periods and 61 controls without RA were determined，and the results were analyzed comparatively with the fluorescent label method（one-step method） for anti-human IgG APF. Results　The positive rates of both methods had no significant differences before and after inactivating serum complement（P＞0.05）. The positive rates of the 2 methods were significantly different（P＜0.05）. Two-step method surpassed the one-step method in sensitivity（75.0%），specificity（96.7%） and positive predictive value（96.6%） which were higher than those of the one-step method. The APF fluorescence intensity in activity period was higher than that in remission period in the identical patients with RA. Conclusions　Two-step method can detect APF antibody of IgM，IgA and IgG types simultaneously，and surpasses the one-step method in resolution，sensitivity and specificity. With rheumatoid factor（RF） and anti-keratin antibodies（AKA），they can complement each other in diagnosis of RA and improve the diagnosis of RA.

Result analysis on anti-HCV reaction determination by ELISA reagents

FU Liqiang;SANG Lieyong;JIANG Guojin

2012, 27(7):  588-591. 

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Objective　To evaluate the efficiency of imported anti-hepatitis C virus（HCV） enzyme-linked immunosorbent assay（ELISA） reagent in blood screening test and analyze the correlations among imported ELISA reagent，domestic ELISA reagent and recombinant immunoblot assay（RIBA）. Methods　A total of 56 anti-HCV positive specimens tested by imported ELISA reagent were retested by 2 domestic ELISA reagents，and the confirmatory test RIBA was also carried out. Results　Among 56 positive specimens with imported anti-HCV ELISA reagent，the confirmatory test RIBA showed that the 12 specimens were confirmed to be positive ，18 specimens were indeterminate and 26 specimens were negative. Of the 9 specimens which were positive with 3 ELISA reagents，7 specimens were confirmed to be positive，1 specimen was indeterminate and 1 specimen was negative. Conclusions　The false positivity with anti-HCV ELISA reagent is obvious，and there would be some different results with different anti-HCV ELISA reagents. For ensuring the blood safety，the imported ELISA reagent and the domestic ELISA reagent should be used simultaneously in anti-HCV screening test.

A retrospective descriptive study on the significance of carbohydrate antigen 125 in differential diagnosis of high and low grade serous ovarian cancer

MAO Peimin;LIU Xiaoyan

2012, 27(7):  592-594. 

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Objective　To investigate the differential diagnosis significance of high and low grade serous ovarian cancer with carbohydrate antigen 125（CA125）. Methods　A total of 128 patients with ovarian tumors were retrospectively analyzed. The serum levels of CA125 of patients with high and low grade serous ovarian cancer and cystadenocarcinoma were determined preoperatively. Results　Serum levels of CA125 among the high and low grade serous ovarian cancer patients were significantly higher than those of serous cystadenocarcinoma patients，respectively（P＝0.002，P＝0.003）. The serum levels of CA125 between the high and low grade serous ovarian cancer patients showed no statistical significance（P＝0.426）. Conclusions　Serum CA125 plays an important role in the differential diagnosis of malignant ovarian tumors，but it cannot identify the high and low grade serous ovarian cancer.

Clinical significance analysis of determination of serum IL-2，4，6，10，TNF-α and IFN-γ in ITP patients

LI Shouwei;LI Xia

2012, 27(7):  595-597. 

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Objective　To evaluate the changes of serum interleukin（IL），tumor necrosis factor-alpha（TNF-α） and gamma-interferon（IFN-γ） in idiopathic thrombocytopenic purpura（ITP） patients before and after treatment. Methods　The concentrations of serum IL-2，4，6，10，TNF-α and IFN-γ were respectively measured by enzyme-linked immunosorbent assay（ELISA） in 53 ITP patients before and after treatment and 50 healthy controls. The results were analyzed. Results　The serum IL-2，IL-6，TNF-α and IFN-γ levels in 53 ITP patients before the treatment were significantly higher than those after the treatment and in the control group.（P＜0.05，P＜0.01）. However，the serum IL-4 and IL-10 levels before the treatment were significantly lower than those after the treatment and in the control group（P＜0.01）. The serum IL-6 levels in 53 ITP patients after the treatment were higher than those in the control group，and the other parameters in 53 ITP patients after the treatment showed no significant difference with those in the control group（P＞0.05）. Conclusions　ITP is characterized by the abnormal presence of multiple cytokines，as an autoimmune disease.The concentration of multiple cytokines can return to be normal after effective treatment.The serum concentrations of IL，TNF-α and IFN-γ in ITP patients should be regularly observed，in order to determine changes in their condition，treatment efficiency and prognosis.

Changes of the number of atypical lymphocyte and T-lymphocyte subsets in children with acute rotavirus diarrhea

YANG Zhen;PAN Ming;ZHAO Haijun

2012, 27(7):  598-600. 

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Objective　To investigate dynamic changes of the number of peripheral blood atypical lymphocyte and T-lymphocyte subsets in children with acute rotavirus（RV） diarrhea. Methods　A total of 40 children with acute RV diarrhea and 20 healthy controls were enrolled in this study. The proportions of peripheral blood atypical lymphocytes in their peripheral blood smear with Wright stain were figured out by microscopy. Meanwhile，the T-lymphocyte subsets were examined by flow cytometry. Results　The proportion of peripheral blood atypical lymphocytes was 16.18%±3.59% in children with acute RV diarrhea and had an obviously increase compared with that in healthy controls（P＜0.001）. The percentages of CD3⁺，CD4⁺ and CD8⁺T cells in peripheral blood of children with acute RV diarrhea and CD4⁺/CD8⁺ ratio were 50.93%±5.86%，22.02%±5.17%，27.07%±4.79% and 0.84%±0.29% respectively. Compared with the healthy controls，the infected groups had a decreased percentage of T cells（P＜0.01）. Conclusions　There are the increasing of atypical lymphocyte number and atypical T-lymphocyte subsets in children with acute RV diarrhea. The determination of atypical lymphocyte number and T-lymphocyte subsets could satisfactorily reflect the immunological state in children with acute RV diarrhea.

Research on correlation of bone marrow CD4⁺CD25^highFoxp3⁺ regulatory T cells with childhood acute B lymphoblastic leukemia

JIANG Limin;XU Chong;FU Qihua

2012, 27(7):  601-605. 

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Objective　To analyze the forkhead box protein 3（Foxp3） mRNA expression and the CD4+ regulatory T cells（Treg） frequency in the bone marrow of childhood acute B lymphoblastic leukemia（B-ALL），and to investigate the relationship between Treg and the development and prognosis of childhood B-ALL. Methods　The levels of Foxp3 mRNA were detected in 35 children with B-ALL，68 patients with remission and 10 healthy controls by real-time polymerase chain reaction（PCR）. The CD4⁺ CD25^highFoxp3⁺ Treg were identified by flow cytometry. The interleukin-10（IL-10） expression level in bone marrow was detected by enzyme-linked immunosorbent assay（ELISA）. Results　The level of Foxp3 mRNA expression in B-ALL group was 4.44 times higher than that of control group. In minimal residual disease（MRD） positive group，it was 2.39 times higher than that in MRD negative group. The percentage median（M） of Treg in CD4+T cells was 12.77% in B-ALL group，which was significantly higher than that in control group（8.09%，P＜0.01）. In addition，Treg frequency in MRD positive group（M＝14.74%） was significantly different from that in MRD continuously negative group（M＝11.30%，P＜0.05）．The level of IL-10 in MRD positive group was（24.69±11.73）pg/mL，which was higher than that of MRD continuously negative group[（9.19±5.82）pg/mL，P＜0.05]. Conclusions　The frequency of Treg and Foxp3 mRNA are significantly elevated in patients with childhood B-ALL and patients with positive MRD，and the IL-10 level in bone marrow is consistent with the frequency of Treg. Treg can be used to evaluate the prognosis and relapse of childhood B-ALL，and may play its role by secreting inhibitory cytokine IL-10.