Table of Content

30 March 2015, Volume 30 Issue 3

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Orginal Article

Introduction of the ICSH guidelines for the verification and performance of automated cell counters for body fluids, 2014 edition

HU Xiaobo, YAO Yiting, WANG Qing, XIONG Lifan

2015, 30(3):  209-213.  DOI: 10.3969/j.issn.1673-8640.2015.03.001

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This article introduces the 2014 edition of International Committee for Standardization in Hematology(ICSH) guidelines for the verification and performance of automated cell counters for body fluids. This guideline provides the particular methods and requirements for laboratories to verify the performance of automated cell counters in the aspects of accuracy, precision, patient correlation, carryover, limits of detection, analytical measurement range, etc. Meanwhile they specify that laboratories should execute the verification of their automated cell counters for different body fluids.

Analysis on the positive rate of M2 anti-mitochondrial antibodies in physical examine screening in Shanghai

CAI Yiting, HUANG Sheng, XIE Songye

2015, 30(3):  214-218.  DOI: 10.3969/j.issn.1673-8640.2015.03.002

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To investigate the early screening significance of M2 anti-mitochondrial antibodies(AMA-M2) for primary biliary cirrhosis(PBC) in physical examine, through analyzing the positive rate of AMA-M2.

Serum samples were collected from 6 658 healthy subjects(3 845 males and 2 813 females), and the sex ratio was 1.37∶1. Their age was 46±13. AMA-M2 was quantitatively determined by enzyme-linked immunosorbent assay(ELISA). Serum alanine aminotransferase(ALT), aspartate aminotransferase(AST), alkaline phosphatase(ALP) and gamma glutamyltransferase (GGT) were determined, and the results were analyzed cemparatively.

AMA-M2 were detected in 97 of 6 658 healthy subjects(1.46%), including 44 males (1.14%) and 53 females(1.88%), and the sex ratio was 1∶1.2. AMA-M2 positive rate of females above 46 years old was higher than those below 46 years old (P<0.01). The 8 of 97 AMA-M2 positive cases without PBC clinical feature had increasing ALP and GGT.

Physical examination should pay attention to the correlation analysis of AMA-M2 with ALP/GGT and ALT with AST. AMA-M2 screening is an effective mean to high risk population in the early detection of PBC. According to American Association for the Study of Liver Disease (AASLD) PBC liver disease diagnostic criteria, positive AMA-M2 and serum liver enzyme help with improving the early detection rate of PBC.

Investigation on the results of thyroid hormone and thyroid autoantibody determinations in 6 112 healthy subjects in Shanghai

YUAN Shuai, JIANG Lu, ZHU Li, LU Longkun, MA Jinlong, WU Yu

2015, 30(3):  219-223.  DOI: 10.3969/j.issn.1673-8640.2015.03.003

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To investigate the situation of thyroid dysfunction among physical examination subjects in Shanghai, and analyze the necessity of thyroid function test in healthy physical examination.

A total of 6 112 subjects in Shanghai undergoing healthy physical examination, including serum thyroid hormone and thyroid autoantibody determinations, were enrolled. Their fasting venous blood samples were collected at early morning. Serum triiodothyronine(T3),thyroxin(T4),free triiodothyonine(FT3),free thyroxin(FT4),thyroid stimulating hormone(TSH),anti-thyroglobulin antibody(TGAb) and thyroid peroxidase antibody(TPOAb) were detected by SIEMENS ADVIA CENTAUR XP automatic chemiluminescence analyzer. The results were analyzed statistically.

Among the 6 112 subjects, the detection rates of hyperthyroidism, hypothyroidism, subclinical hyperthyroidism and subclinical hypothyroidism were 0.47, 0.34, 0.62 and 2.55, respectively. Female incidence rates of hyperthyroidism, hypothyroidism, subclinical hyerthyroidism and subclinical hypothyroidism were higher than those in males(P=0.001). Female abnormal rates of serum thyroid hormone and thyroid autoantibodies were significantly higher than those in males(P=0.001). Serum thyroid hormone in 10.37% of the subjects was normal, however, with the increasing levels of thyroid autoantibodies. With the increasing of age, the incidence rate of thyroid dysfunction and the abnormal rates of serum thyroid hormone and thyroid autoantibodies showed a trend of increase.

Hypothyroidism and subclinical hypothyroidism are major thyroid dysfunctions in Shanghai, with higher female incidence and abnormal rates than male's, and there is a relationship with age. The determinations of thyroid hormone and thyroid autoantibodies in healthy physical examination are very necessary and valuable.

The analysis of thyroid hormone levels of different pregnant trimesters

KE Wencai, GU Yunxia

2015, 30(3):  224-226.  DOI: 10.3969/j.issn.1673-8640.2015.03.004

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To investigate the thyroid related hormones of differenct pregnant trimesters and put forward suggestions about related routine tests.

1 500 cases of pregnant women at different trimesters were retrospectively analyzed, including 500 cases in first trimester(<12 weeks), 500 cases in second trimester(13-27 weeks) and 500 cases in third trimester(28 weeks to delivery). 200 healthy non-pregnant women were selected as control group. The serum thyroid hormone levels were tested, such as free 3,5,3'-triiodothyronine(FT3), free thyroxine(FT4), thyroid stimulating hormone(TSH), thyroglobulin antibody(A-TG) and anti-thyroid peroxidase antibody(TPOAb). Statistical analysis were performed.

There was statistical significance between the pregnant group and control group in terms of serum thyroid hormones levels(P<0.05). The morbidity rate of pregnant group was higher(8.53%) than that of control group. The difference among the different trimesters showed statistical significance(P<0.05). From first to third trimesters, TSH levels increased gradually while FT3, FT4 decreased gradually. The positive rates of ATG and TPOAb were 4.6% and 3.6%, respectively for the third trimester, which were lower than first(12.4%, 11.4%) and second trimester groups(9.0%, 8.2%).

There is close relationship between serum thyroid hormones level and trimesters. The pregnant women should be screened for serum thyroid hormones levels and intervened if necessary, in order to achieve better delivery.

Correlation research on serum ferritin and homocysteine levels with carotid atherosclerosis in essential hypertension patients

YANG Wendong

2015, 30(3):  227-229.  DOI: 10.3969/j.issn.1673-8640.2015.03.005

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To investigate the correlations of serum ferritin (SF) and homocysteine (Hcy) levels with carotid atherosclerosis (CAS)in patients with essential hypertension (EH).

SF and Hcy levels in 266 patients with EH were determined by electrochemiluminescence immunoassay and circulating enzymatic method. According to carotid intima-media thickness (CIMT, color doppler ultrasound instrument), 266 patients with EH were classified into CIMT thickening group (167 cases, CIMT>1.0 mm) and CIMT non-thickening group (99 cases, CIMT≤1.0 mm). A total of 50 healthy subjects were enrolled as control group.

SF and Hcy levels in CIMT thickening group were significantly higher than those in CIMT non-thickening group(t=12.561 and 4.581, P<0.01). SF and Hcy levels in CIMT non-thickening group were higher than those in control group (t=6.305 and 2.913, P<0.01). SF and Hcy levels in CIMT thickening group were positively correlated with CIMT (r=0.597 and 0.451, P<0.01).

EH patients have high levels of SF and Hcy, and SF and Hcy levels are correlated to EH combined with CAS. The combined detection of SF and Hcy levels contributes to the condition and prognosis judgement.

The significance for thyroglobulin detection in suspicious cervical lymph node fine-needle aspirate washout fluid after surgery in papillary thyroid carcinoma patients

MAO Minjing, YE Tingjun, SHI Xinming, PENG Yibing, WANG Xuefeng

2015, 30(3):  230-233.  DOI: 10.3969/j.issn.1673-8640.2015.03.006

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To determine the change of thyroglobulin(Tg) in fine-needle aspirate(FNA)of cervical lymph node(CLN) in order to provide the reference of papillary thyroid carcinoma(PTC) metastasis identification in lymph node Under ultrasound-guided FNA, 109 samples from patients that were suspicious for metastatic PTC were collected, and 16 samples from patients whose cervical lymph nodes diagnosed as reactive were used as control group. FNA-Tg levels were measured by immunochemiluminescence assay. Then, the relationships between FNA-Tg levels and metastatic PTC were investigated. The positive detection rates of FNA-Tg test and traditional cytology test were compared. The macrophage cell and tumor cell in FNA slides were counted, then divided them into 4 groups.The rank sum test was used to analyze the differences of FNA-Tg levels from samples of different cellular components.

Among the 109 samples, 84 samples had positive FNA-Tg results which were consistent with their pathological results(metastatic PTC). The detect rates of traditional cytology test in simple metastatic PTC and cystic metastatic PTC were 93.9% and 88.6%, respectively. However, the detection rates of FNA-Tg test were 100% in both simple and cystic metastatic PTC. After group clavsification, according to their cellular components, is showed that FNA-Tg levels were not associated with the number of tumor cells. FNA-Tg levels of the group with cystic macrophage were significantly higher than those of the group with no macrophage(U=367.00,P<0.01).

Tg test by FNA washout fluid can be used as a valuable marker of lymph node metastasls from PTC. FNA-Tg test could improve the deteetiom sensitivity in diagnosing metastatic PTC especially in cystic metastasis. Macrophages seen in traditional cytology test could be indicative of metastasis.

The clinical significance of ischemia modified albumin and cardiac troponinⅠcombined determination in the diagnosis of acute coronary syndrome

YU Pingan, PAN Feng

2015, 30(3):  234-237.  DOI: 10.3969/j.issn.1673-8640.2015.03.007

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To investigate the clinical significance of ischemia modified albumin (IMA) and cardiac troponinⅠ (cTnI) combined determination in the diagnosis of acute coronary syndrome (ACS).

A total of 127 patients with ACS were enrolled, including 49 patients with unstable angina pectoris(UAP), 40 patients with ST elevation acute myocardial infarction (STEMI) and 38 patients with non-ST elevation acute myocardial infarction (NSTEMI). In the same period, 60 healthy controls were enrolled. The cTnI concentration in patients with ASC for the first-time determination and IMA concentration by enzyme-linked immunosorbent assay (ELISA) to detect the same sample retrospectively were taken.

Compared with control group, IMA concentrations in UAP, STEMI and NSTEMI groups were higher (P<0.05), those in STEMI and NSTEMI groups increased compared with that in UAP group (P<0.05), and that in STEMI group significantly increased compared with that in NSTEMI group (P<0.05). The cTnI comcentrations in STEMI and NSTEMI groups were higher than that in UAP group (P<0.05), that in STEMI group had no difference with that in NSTEMI group (P>0.05). IMA and cTn Ⅰ expression existed positive correlation (r=0.536,P<0.05). Receiver operating characteristic (ROC) curve showed the area under the ROC curve of IMA in the diagnosis of ACS was 0.87±0.028, the corresponding sensitivity 92%, the specificity 78%, while the area under the ROC curve for cTnI was 0.74±0.021, the corresponding sensitivity 77%, the specificity 91%, and there was statistical significance (P<0.05).

IMA and cTnI assays for the diagnosis of myocardial infarction has important clinical value, and IMA diagnosis compared with cTnI is more ideal.

Early diagnosis significance of immature granulocyte flag in patients with sepsis

WANG Lejian, SHI Chunjuan, LI Fei, ZHANG Xiwen

2015, 30(3):  238-239.  DOI: 10.3969/j.issn.1673-8640.2015.03.008

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To investigate the early diagnosis significance of immature granulocyte (IG) in patients with sepsis.

A total of 185 in patients with suspected sepsis in our hospital between January 2008 and June 2013 were enrolled. Their relative data were analyzed retrospectively. The blood culture and peripheral blood IG warning flag results were analyzed statistically.

Among the 185 cases, there were 134 cases of blood culture positive. There were IG ≤0.50% (7 cases), IG ≤3.00% (32 cases), IG ≤4.00% (65 cases) and IG >4.00% (30 cases). Blood culture negative had 51 cases in the 185 cases, and IG ≤0.50% (26 cases), IG ≤3.00% (17 cases), IG ≤4.00% (5 cases) and IG >4.00% (3 cases). It was significant between IG >3.00% group and the other groups(P<0.05), when the results of blood culture was positive. It was significant between IG ≤3.00% group and the other groups(P<0.05), when the results of blood culture was negative.

The results of IG >3.00% can be used as a early auxiliary diagnosis index for patients with sepsis. It has specificity in the diagnosis of sepsis.

The comparability verification for the results of different automated hematology analyzers

ZHU Xiang, HAN Lingfei, XIE Ping, ZHAO Weisong

2015, 30(3):  240-242.  DOI: 10.3969/j.issn.1673-8640.2015.03.009

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To analyze comparatively for white blood cell (WBC), red blood cell (RBC) and platelet (PLT) results of different automated hematology analyzers, to evaluate the consistency, in order to provide reliable results for clinic.

According to THE Guideline for Comparability Verification of Quantitative Results within One Health Care System, the WBC, PLT, RBC, HGB, HCT, MCV, MCH and MCHC were detected by different automated hematology analyzers, and their relative bias (R) was calculated. As the standard of 1/3 allowed total error (TEa) of Clinical Laboratory Improvement Amendments (CLIA)'88, the comparability among different analyzers was analyzed.

The R of WBC, PLT, RBC, HGB, HCT, MCV, MCH and MCHC by different automated hematology analyzers were <1/3 CLIA'88 TEa, which were all acceptable.

The results of WBC, PLT, RBC, HGB, HCT, MCV, MCH and MCHC by different analyzers have good consistency and comparability which is suitable for clinic.

Automatic verification procedure and evaluation of Mindray CAL 8000 auto sample processing line

WEI Jian, WANG Jianbiao, SONG Weixing, CHEN Liting, SHI Hourong, DONG Shuzhen, LU Ying

2015, 30(3):  243-246.  DOI: 10.3969/j.issn.1673-8640.2015.03.010

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To establish the automatic verification rules and validate the automatic verification system of Mindray CAL 8000 auto sample processing line.

1 025 EDTA-K₂ anti-coagulated venous whole-blood samples were collected from Ruijin Hospital(including outpatient and inpatient samples). Each sample was both analyzed on the analyzer in CAL 8000 auto sample processing line and performed manual differential. The analysis results and microscopic results were used to evaluate the drafted automatic verification rules. Furthermore, 40198 outpatient and inpatient sample results obtained from March to May in 2014 were used as experimental data by Mindray automatic verification software(LabXpert), where the percentage of samples passing the automatic verification was obtained and the samples failing in the verification were categorized. A comparison of verification time using the software and in manual way was also performed.

The false negative percentage of 1 025 samples tested was 0.78%, lower than the 5% required by the International Hematology Consensus Group. The false negative items mainly included erythrocyte morphology, platelet clump and toxic granules of neutrophils. By analysis of the 40 198 samples using the automatic verification software, 65.3% samples need automatic verification, and 34.7% need manual verification. Samples requiring manual verification included: PLT out-of-range samples, WBC out-of-range samples, samples from hematology department, samples with immature granulocyte flag, RBC and MCV out-of-range samples, samples with blast cell flag, HGB out-of-range samples, etc.

Using the automatic verification software increases clinical work efficiency, decreases false negative ratio, saves the report time and improves the patients' satisfaction.

Study on the coagulation function in late-pregnancy women with viral hepatitis and its clinical significance

FU Jie, ZHANG Rui, XIE Lili, LI Jianwei, DONG Zhiwu

2015, 30(3):  247-249.  DOI: 10.3969/j.issn.1673-8640.2015.03.011

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To observe the variation of coagulation function and discuss its clinical significance in late-pregnancy women with viral hepatitis.

A total of 134 late-pregnancy women with viral hepatitis were classified into 5 subgroups: acute hepatitis(39 cases), chronic mild hepatitis(43 cases), chronic moderate/severe hepatitis(25 cases), cirrhosis(15 cases) and severe hepatitis(12 cases) groups. The prothrombin time (PT), activated partial thromboplastin time (APTT) and fibrinogen (FIB) were determined and compared with the results of 45 normal late pregnant women. Meanwhile, the blood coagulation factors, including FⅡ, FⅤ, FⅦ, FⅧ, FⅨ, FⅪ and FⅫ, of 23 chronic mild hepatitis patients, 13 chronic moderate hepatitis patients, 10 cirrhosis patients and 26 normal late pregnant women, were measured respectively.

Compared with normal late pregnancy group, PT and APTT were prolonged in severe hepatitis group (P<0.05). FIB decreased in both cirrhosis group and severe hepatitis group (P<0.05, P<0.01). FⅦ:C in chronic mild hepatitis and chronic moderate hepatitis groups were lower than that in normal pregnant group. Most coagulation factors, except FⅧ:C, in cirrhosis group were lower than those in the normal late-pregnancy group (P<0.05).

Coagulation function in late-pregnancy women is affected with viral hepatitis. It is of great clinical significance to determine the coagulation indices for preventing postnatal hemorrhea and other complications.

Evaluation on the performance of a latex agglutination assay for treponemal specific antibody

HU Liang, CHEN Yi, CHEN Changqiang, QU Chenyun, GU Zhidong, FAN Qishi

2015, 30(3):  250-253.  DOI: 10.3969/j.issn.1673-8640.2015.03.012

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To evaluate the analytic performance of a latex agglutination assay for treponemal specific antibody (Mediace TPLA).

The EP documents established by Clinical and Laboratory Standards Institute (CLSI) were referred to evaluate the analytic performance. A total of 110 syphilitic positive and 78 syphilitic negative sera were detected by Mediace TPLA,and the results were compared with those of chemiluminescence microparticle immunoassay (CMIA).

The total coefficients of variation (CV) were 8.77%, 3.94% and 6.57% respectively at the concentration levels of 8.0, 28.6 and 166.0T.U.. The linear range was from 0 to 250 T.U., and the total recovery rate was 95.31%. The results were not significantly affected by hemolysis (hemoglobin<4.8 g/L), lipidemia (triglyceride<8 mmol/L), jaundice (bilirubin <20 μmol/L) and rheumatoid factor (<450 IU/mL). The results of 30 antinuclear antibody (ANA) positive patients, 10 hepatitis patients and 10 pregnant women were all negative. When the results of CMIA were defined as the reference standard,the sensitivity of Mediace TPLA was 99%, the specificity was 100%, and the cut-off value was 10 T.U..

Mediace TPLA has excellent anti-interference ability, repeatability, high accuracy, wide linear range and specificity. It can also be used in automated biochemical analyzers.

Study on the diagnostic significance of T cells spot test in Mycobacterium tuberculosis infected patients with pleural effusion

XU Xuecheng, MAO Liming

2015, 30(3):  254-257.  DOI: 10.3969/j.issn.1673-8640.2015.03.013

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To investigate the application significance of Mycobacterium tuberculosis (MTB) T cells spot test (T-SPOT) in the rapid diagnosis for suspected MTB infected patients with pleural effusion.

The 124 patients with pleural effusion which were suspected as having suspected MTB infection were enrolled. T-SPOT was used to detect the immune response of peripheral blood lymphocyte against MTB special secreted protein EAST-6 and CFP-10. The MTB DNA method was used to detect MTB in pleural effusion, meanwhile serum anti-MTB antibody was detected by immuno-gold filtration assay, and the results were analyzed statistically.

The 44 of 124 cases were diagnosed as tuberculosis by sputum, pleural effusion and ascites smear test, sputum, pleural effusion and ascites MTB culture, tuberculin test, MTB DNA method and anti-MTB antibody test, and the positive rate was 35.5%. The positive rate of T-SPOT in the 124 patients with pleural effusion was 33.06%(41/124). The positive rate of MTB DNA method was 14.52%(18/124), and that of anti-MTB antibody test was 9.68%(12/124). Among the 44 confirmed tuberculosis patients, T-SPOT had the consistency rate of 86.36%(38/44), while the consistency rates of MTB DNA method and anti-MTB antibody test were 27.27%(12/44) and 18.18%(8/44), respectively. The positive rate and consistency rate of T-SPOT were significantly higher than those of the others (P<0.05). The positive and negative predictive values of T-SPOT in suspected MTB infected patients with pleural effusion were 92.68% and 92.77%, respectively.

T-SPOT is fast, sensitive and specific to diagnose the suspected MTB infected patients with pleural effusion. T-SPOT is better than MTB DNA method and anti-MTB antibody test. T-SPOT can become a new diagnostic method to be applied in the diagnosis of MTB infection.

The clinical significance of smear and Gram staining for microscopy before microbial specimens being cultured

ZHANG Guoying, XIA Xuehong

2015, 30(3):  258-261.  DOI: 10.3969/j.issn.1673-8640.2015.03.014

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To investigate the clinical significance of microbial specimens being smeared and Gram stained for microscopy before they were cultured .

A total of 365 clinical specimens with different types of microorganisms were firstly smeared and stained for microscopy, then were inoculated for culture. Finally, the coincidence rate was analyzed.

Among the 365 specimens, there were 211 sputum specimens, 40 urine specimens, 76 secretion specimens and 38 other-type specimens (mainly pus and pleural effusion specimens). Among them, there were 17 unqualified specimens. Of the 348 qualified specimens, 93 cases were cultured positive. The detection results of sputum, urine, secretion and other-type specimens were no statistical significance by the smear microscopy and the culture method. The coincidence rates were 76.6%, 62.5%, 87.1% and 80.0%, respectively (P>0.05).

The smear microscopy can not only check the quality of specimens, but also reduce the result errors of culture, so as to improve the positive rate of specimens by culture. Therefore, it is of great significance in the actual microbiological testing work.

Analysis on the in vitro susceptibility test results from two groups of traditional Chinese medicine prescriptions for diabetic foot producing ESBLs Escherichia coli and MRSA

LIANG Xiaoyi, ZHU Haiying, LIU Xiaomei, PAN Min

2015, 30(3):  261-264.  DOI: 10.3969/j.issn.1673-8640.2015.03.015

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To understand the inhibitory and treatment effects of Coptis chinensis and Coptis chinensis with Musk prescriptions for diabetic foot producing extended-spectrum beta lactamase (ESBLs) Escherichia coli and methicillin-resistant Staphylococcus aureus (MRSA).

The patients diagnosed as diabetic foot were enrolled. After culturing, 138 isolates of bacteria were isolated, and 67 isolates of ESBLs Escherichia coli and MRSA were screened. The minimum inhibitory concentrations (MIC) of Coptis chinensis (prescription 1) and Coptis chinensis with Musk (prescription 2) were determined by agar dilution method. The treatment effect of the 2 groups of prescriptions for diabetic foot was observed.

The MIC of prescription 1 for 35 isolates of ESBLs Escherichia coli was ≥128 mg/mL, and the MIC of prescription 2 was 32 mg/mL. The MIC scope of prescription 2 was 32-128 mg/mL. The MIC for 32 isolates of MRSA droped from 32 mg/mL (prescription 1) to 8 mg/mL (prescription 2). Compared with prescription 1, the MIC of prescription 2 decreased significantly, the treatment effect was distinct, and the differences were statistically significant (P<0.05).

Traditional Chinese medicine prescription Coptis chinensis with Musk has inhibitory and treatment effects for diabetic foot producing ESBLs Escherichia coli and MRSA, which is a novel method for clinical treatment.

Establishment of fluorescence quantitation PCR for the quantitative detection of five species of Candida

XIA Qianfeng, FU Qiongyao, WU Qiang, QIN Xi, YAO Maozhong, HUANG Mianqing, XING Guilan, QIAN Shiyun

2015, 30(3):  265-268.  DOI: 10.3969/j.issn.1673-8640.2015.03.016

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To establish and evaluate fluorescence quantitation polymerase chain reaction (PCR) for the quantitative detection of 5 species of Candida.

The presence of DNA from pathogens in the Candida species was detected by the PCR targeting internal transcribed spacer region of fungal gene. The PCR was optimized and evaluated methodologically.

The fluorescence quantitation PCR showed a wide range of linearity from 10¹-10⁹ CFU/mL, high sensitivity 5 CFU/mL, within-run and between-run coefficients of variation (CV) <5%. When the PCR was applied directly to identify 19 mocked double-blind blood samples, 5 of them were positive to 5 species of Candida, and the other 14 of them were negative to other pathogens.

The PCR may be appropriate for clinical laboratories as accurate, rapid and reliable determination for the most frequently encountered Candida species.

Primary research on freeze drying technology for the preparation of coagulation factor external quality control material in local area

SONG Ying, DAI Jing, WANG Qing, DING Qiulan, WANG Xuefeng

2015, 30(3):  269-273.  DOI: 10.3969/j.issn.1673-8640.2015.03.017

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To use freeze drying technology for preparing plasma coagulation factor external quality control material, in order to use it into local quality control scheme.

Fresh plasma was treated with freeze drying protective agent. Filling the plasma in vials, Germany Christ freeze dryer was used to lyophilize the plasma. The self-made materials were prepared, which should meet the requirements of Guidance on Evaluating the Homogeneity and Stability of Samples Used for Proficiency Testing (CNAS-GL03). The materials were deliveried into several hospitals for the determination of coagulation factor.

By screening tests and orthogonal experiments, the optimum freeze drying protective agent's formula was trehalose (1.5%), glycine (0.5%) and HEPES (0.2%). The average packing bottle precision value was 0.569 9 g (net weight), ranging from 0.564 6 g to 0.576 7 g, S was 0.003 5 g, and coefficient of variation (CV) was 0.61%. The average moisture of self-made lyophilized product was 1.22%, S was 0.197%, and CV was 16%. Homogeneity testing used 2 methods to evaluate (1) F test: F=0.06; (2) T test: P=0.053 so the homogeneity met the requirements of CNAS-GL03. The stability testing used |\begin{array}{c}\stackrel{\text{?}}{x}\end{array}-\begin{array}{c}\stackrel{\text{?}}{y}\end{array}|≤0.3σ, and the absolute value of difference was 0.07, which was < critical value 1.702, which met the CNAS-GL03 requirements. By using self-made lyophilized product in external quality control survey, the results of coagulation factor Ⅷ CV% were 21.3, 16.6 and 25.8(the results were similar to those of the National Center for Clinical Laboratory).

The self-made coagulation factor external quality control material can meet the requirements of external quality control scheme in local area.

Comparison and application of low platelet counts by CD61 immunological, optical, electrical impedance methods of CELL-DYN Sapphire hematology analyzer

ZHANG Chi, ZHANG Hongbo

2015, 30(3):  274-279.  DOI: 10.3969/j.issn.1673-8640.2015.03.018

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To evaluate and compare the advantages and disadvantages of 3 detection methods of CELL-DYN Sapphire hematology analyzer in the aspect of low platelet (PLT) detection.

A total of 100 patients whose platelet <50×10⁹/L caused by basis hematonosis or after chemotherapy were enrolled. PLT counts were determined by electrical impedance method (IPLT), optical method (OPLP) and CD61 immunological method (CD61-PLT). The results were analyzed comparatively with those of manual microscopy. Variance analysis, Passing-Bablok regression analysis and Bland-Altman bias analysis were performed by SPSS 19.0 and MedCalc V12.7.2.0 softwares statistically.

ANOVA analysis showed that there was statistical significance for IPLT and OPLT with manual method (MPLT). (P=0.00, P=0.002). CD61-PLT and MPLT had no statistical significance (P=0.915). OPLT and CD61-PLT had good correlation with MPLT without statistical significance[slope 1.0, 95% confidence interval (CI):0.95-1.06, r=0.946 and slope 1.0, 95%CI: 0.99-1.01, r=0.998]. IPLT had poor correlation with MPLT with statistical significantce (slope 1.27, 95%CI: 1.10-1.44, r=0.845). According to variance analysis, the values of IPLT and OPLT were higher than those of MPLT (mean deviations were 6.3 and 1.3). CD61-PLT and MPLT had no significant difference(mean deviation was -0.02).

IPLT has significantly statistical difference with MPLT for low PLT determination, with poor correlation, and the result has a significant upward deviation. The means of OPLT and MPLT have statistical significance with good correlation, but the result has a still small upward deviation. The CD61-PLT and MPLT have no obvious difference with a good correlation, and the results are not significantly different. Therefore, it is recommended that we can use CD61-PLT as a new reference method and OPLT and IPLT as alternative methods.

The influence of beta-glucan on TNF-α,IL-8 and IL-12 in monocytes

WU Rongqiang, WANG Sujian, WANG Shizhong, ZHOU Haijun, QI Chunjian

2015, 30(3):  280-283.  DOI: 10.3969/j.issn.1673-8640.2015.03.019

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To study the influence of beta-glucan on tumor necrosis factor alpha (TNF-α), interleukin 8 (IL-8) and interleukin 12 (IL-12) in atherosclerosis pathogenesis monocytes.

The monocytes of healthy subjects were separated by density gradient centrifugation. The monocytes were induced into macrophages by phorbol myristate acetate (PMA), and the macrophages were induced into foam cells by oxidized low-density lipoprotein (ox-LDL). There were control group, ox-LDL group, whole beta-glucan particle (WGP) group and WGP + ox-LDL group, the total RNA was extracted, and the TNF-α, IL-8 and IL-12 cDNA was amplified. The concentrations in supernates were determined by enzyme-linked immunosorbent assay(ELISA).

Compared with control group,the protein expression of TNF-α ,IL-8 and IL-12 in monocytes were significantly up-regulated by ox-LDL (P<0.05). After pretreated with WGP,the protein expressions of TNF-α, IL-8 and IL-12 and the mRNA expressions of TNF-α and IL-8 were inhibited significantly (P<0.05).

beta-glucan may inhibited ox-LDL-induced pro-inflammatory effects in macrophages,which attenuates the development of atherosclerosis.

Result analysis of various detection methods for homocysteine

ZHAO Yan, KANG Fengfeng, WANG Wei, HE Falin, ZHONG Kun, WANG Zhiguo

2015, 30(3):  284-286.  DOI: 10.3969/j.issn.1673-8640.2015.03.020

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To evaluate the results of various detection methods for homocysteine (Hcy).

The internal quality control (IQC ) information of 110 laboratories from National Center for Clinical Laboratory (NCCL) was collected. The imprecision [coefficient of variation (CV)] of Hcy was analyzed. Bias was estimated according to various detection methods in NCCL External Quality Assessment (EQA) in 2012 through online EQA platform. With current evaluation criteria for EQA(TEa=20%), sigma (σ) value of each laboratory was calculated as the formula: σ= (TEa-bias)/CV. Furthermore, the passing rates and inter-laboratory CV for each group were evaluated.

The proportions of σ>6, 3<σ≤6, 0<σ≤3 and σ≤0 by chemiluminescence, immunoturbidimetry and enzymatic cycling were 0%, 7.69%, 30.77%, 61.54%;9.09%, 0%, 63.64%, 27.27% and 11.63%, 31.40% 47.67%, 9.30%, respectively. The inter-laboratory CV by the 3 methods for 5-run quality control materials were 33.66%-43.77%, 19.54%-25.24% and 10.46%-14.21%, and the passing rates were 53.3%, 53.3%-80.0% and 85.9%-92.3%.

Laboratories in China need further efforts to improve the performance of Hcy detection.

The root cause analysis of common nonconformity in medical laboratory accreditation

GUO Xiaojun, HUANG Weigang

2015, 30(3):  295-297.  DOI: 10.3969/j.issn.1673-8640.2015.03.025

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By comparing the distribution of non-conformity clauses in the activity of medical laboratories accreditation,it was found that the first six clauses differed between our investigated 18 medical laboratories in Shanghai and the 31 ones nationwide from 2004 to 2008. In order to improve the quality and provide valuable information for laboratory accreditation, we analyzed root cause of nonconformity items that happen most in frequency, and the difference with the efforts to find the reasons, referred to the advanced experiences from overseas on quality management and then discussed the effective measures to make progress.

The immunosuppression of lymphocyte activation gene 3 and its effects in chronic viral infection

MA Chenyun, LU Zhicheng, SHEN Qian

2015, 30(3):  298-302.  DOI: 10.3969/j.issn.1673-8640.2015.03.026

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T cell depletion is the main mechanism that T cell responses can not effectively control viral replication and eliminate virus. Lymphocyte activation gene 3 (LAG-3) is a vital immune co-inhibitory molecule which is similar to CD4 molecule structure and mainly expresses on activated T cells. LAG-3 can be combined with major histocompability complex (MHC)-Ⅱ molecules, and it's also a negative regulator of T lymphocyte's proliferation and function for maintaining the homeostasis. LAG-3 highly expresses on the surface of virus-specific T cells causing T cell depletion in chronic viral infection.