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Effect of LIF gene modified ECV-304 cells on the ex vivo culture of HSC/HPC in cord blood

Chinese Journal of Clinical Laboratory Science.2006;0(02):-.

Objective To study the effect of the ECV-304 cells modified with LIF gene on the ex vivo culture of HSC/HPC in cord blood.Methods The ECV-304 cells were infected by Eukaryotic Expression plasmid pcDNA3.0LIF,and the positive ECV-304 cells were obtained by selected with G418.These cells were used to co-culture with CD34+ cells of cord blood.The phenotype of CD34+ and CD34+ CD54+、CD34+ CD62L+ primitive progenitors was detected by flow cytometry.Results The LIF gene can express in ECV-304 cells steadily.ECV-304 cells modified with LIF gene can improve expansion of CB CD34+、CD34+CD54+ and CD34+ CD62L+ cells while sustaining the expression of homing-related adhesion molecule.Conclusion The ECV-304 cells modified with LIF gene can not only significantly expand CB hematopoietic progenitor cells ex vivo,but the expanded CD34+ cells may well retain their homing ability.

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Verification and application of genetic screening processor(GSP) for neonatal screening

Yanmin WANG ; Guoli TIAN ; Zhuo ZHOU ; Jing GUO ; Hongping XU ; Jing YAO

Chinese Journal of Clinical Laboratory Science.2017;35(2):152-154. doi:10.13602/j.cnki.jcls.2017.02.20

Objective To evaluate the feasibility of PerkinElmer Genetic Screening Processor(GSP) in the application of newborn screening for congenital hypothyroidism (CH) and congenital adrenal hyperplasia (CAH) by detecting thyroid-stimulating hormone (TSH) and 17-OH-progesterone(17-OHP).Methods The dried-blood spots specimens from Centers for Disease Control and Prevention(CDC) and the quality control in the reagent kit were detected and the accuracy,precision and linearity were calculated.A total of 1 012 samples of TSH(60 of positive and 952 negative samples) and 991 samples of 17-0HP(34 positive and 957 negative samples)were detected.The initial cut-off value was determined by ROC curve determined.The consistency between the results from GSP and clinical diagnosis was analyzed.Results The average of within-run coefficient of variation(CV) of TSH and 17-OHP were 6.69％ to 12.6％ and 7.52％ to 9.29％,and the average of between-run CV were 6.91％ to 10.96％ and 6.86％ to 12.36％,respectively.The average of bias of TSH and 170HP were-14.28％ to-0.74％ and-0.45％ to 12.54％.The linearity of GSP detection was fine.The initial cut-off values were 23.43 U/mL(TSH) and 21.42 ng/mL(17-OHP).The sensitivity of GSP detection was 100％ and the specificity of TSH and 17-OHP were 98.11％ and 99.58 ％ respectively.The results of GSP detection showed good consistency with clinical diagnosis.Conclusion As the first real automatic fluorescence immunoassay analyzer,GSP could be used in routine clinical diagnosis for CH and CAH.

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Selection and evaluation of calibrators for the conventional measurement system of serum α-Amylase

Yan HUANG ; Yu ZONG ; Juan JIAN ; Huaian MA

Chinese Journal of Clinical Laboratory Science.2017;35(2):149-151. doi:10.13602/j.cnki.jcls.2017.02.19

Objective To select the calibrator for the conventional measurement system of serum a-Amylase (Amy).Methods The Amy levels of forty frozen serum samples were detected by the IFCC reference method (reference system),the conventional system A which used the Bioassay routine reagent and Randox calibrator,and was calibrated by the Roche PNPG7 method,and the conventional system B which used the Bioassay routine reagent and Randox calibrator,and was calibrated by the Rondox liquid stable PNPG7 method,respectively,and the acceptability of the two conventional systems was evaluated.Results The regression equations of the measurement values between the IFCC reference method and the conventional systems A and B were Y =0.964X +0.376 and Y =1.095X + 3.131,respectively.Among them,X and Y represented the results of the IFCC reference method and the conventional system,respectively.Compared with the IFCC reference method,the results of the conventional system A was reliable.Condusion With the guidance of the IFCC reference method,the domestic biochemical reagents matched with the suitable calibrators may provide the acceptable results.

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Application of the 5W1H method combined with PDCA circulation in the protection of occupational risks for technicians in blood centers

Yi LIU ; Liqiang FU ; Shoubing ZHU

Chinese Journal of Clinical Laboratory Science.2017;35(2):145-148. doi:10.13602/j.cnki.jcls.2017.02.18

Objective To improve the protective strategy of occupational risks for technicians in blood centers so as to ensure their health and safety.Methods The occupational risk factors were analyzed by the 5W1H method,and the corresponding protective measures were introduced.Then,the protective measures were verified dynamically by the PDCA circulation,and were standardized and further spread.The projects with poor effect were replanned and entered the PDCA recycling.Results The system of the 5W1H method combined with PDCA circulation could identify and control various occupational risks,and effectively prevent the occupational detriments.Condusion Combined application of the 5W1H method with PDCA circulation is an effective way to protect the health and safety of technicians in blood centers and an effective mode to improve the management of occupational risks.

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Retrospective analysis of external quality assessment results for hemoglobin measurement in Guangxi Province during 2012 to 2016

Juan TANG ; Weiya ZHOU ; Xiangyang ZHOU ; Xiaochun LIU ; Yi HE

Chinese Journal of Clinical Laboratory Science.2017;35(2):142-144. doi:10.13602/j.cnki.jcls.2017.02.17

Objective To evaluate the detection competence of HbA2 and HbF in Guangxi medical laboratories.Methods The external quality assessment(EQA) of HbA2 and HbF was conducted twice a year and five samples was detected each time during 2012 to 2016.The laboratories participated in EQA completed the samples' detection and submitted the detection results at specified time according to the requirements of EQA.The distribution of each detection system,the qualification rate of each laboratory,the variation degrees of each detection system and each detection method,and the variations of results for different levels of quality control(QC) materials during 5 years were analyzed based on the returned results.Results The application of high performance liquid chromatography (HPLC) and capillary electrophoresis(CE) increased year by year and their usage rates in 2016 reached up to 46.1％ (82/178) and 18.0％ (32/178),respectively.The qualification rates of HbA2 and HbF increased from 51.5％ (34/66) and 60.6％ (40/66) in 2012 to 93.3％ (166/178) and 92.1％ (164/178) in 2016,respectively.The average coefficient of variation(CV) of each detection system decreased year by year.There were good CVs for the results of high,medium and low levels of HbA2 QC materials detected by the Bio-Rad Variant Ⅱ and Sebia CAPILLARYS 2 systems,and they were less than 6.0％.HPLC and CE could quantitatively detect the HbA2 and HbF levels,and their total detection competence was superior to that of agarose gel electrophoresis.Conclusion EQA can assess the abihty of one laboratory detecting HbA2 and HbF,and quantitatively analyze the levels of HbA2 and HbF,which may provide the quality assurance and data support for the screening and prevention of thalassemia.

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Differential proteomic analysis for immune complexes deposited in synovial tissues of RA patients using liquid chromatography-tandem mass spectrometry

Haiyong WANG ; Xiaolong GUAN ; Qinfei ZHAO ; Yuzhong XU ; Yu ZHANG ; Shusheng SONG ; Ang LI ; Xing LV ; Ying ZHOU ; Wei YU ; Jianning ZHAO ; Xiaojun LI

Chinese Journal of Clinical Laboratory Science.2017;35(1):15-20. doi:10.13602/j.cnki.jcls.2017.01.04

Objective To scan protein expression profile of immune complexes (ICs) derived from the synovial tissue of the patients with rheumatoid arthritis (RA) based on liquid chromatography-tandem mass spectrometry (LC-MS).Methods The samples of synovial fluid were obtained from knee joints of the patients with RA and osteoarthritis (OA) used as control during therapeutic arthrocentesis in knee jiont at the Department of Orthopedics of Jinling Hospital,School of Medicine,Nanjing University.The protein expression profile of ICs was identified by enrichment strategy based on immunoprecipitation and LC-MS analysis.The value of fraction of total (FOT) was used to estimate protein abundance and screen the up-and down-regulated proteins.The function enrichment,interaction network and signal pathway of differential proteins were analyzed using softwares David and String.Results A total of 511 and 526 protein spots in ICs of RA and OA patients were identified respectively.Among them,170 proteins existed only in RA group.45 and 85 proteins in RA group were statistically up-and down-expressed compared with controls.Conclusion HSP90AA1,HSP70,HLAG,Thioredoxin,Annexin A2 and vitronectin may be involved in the pathogenesis of RA through different paths and possible to become promising diagnostic indicators or new therapeutic targets for RA.

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Correlation of baseline neutrophil-to-lymphocyte ratio with the curative effect and prognosis of breast cancer patients after neoadjuvant chemotherapy

Meirong FAN ; Yangtao OU ; Yingjian HE ; Zhexuan LI ; Xueshuo GAO ; Qingyun ZHANG ; Guobin XU

Chinese Journal of Clinical Laboratory Science.2017;35(2):114-117. doi:10.13602/j.cnki.jcls.2017.02.09

Objective To investigate the predictive value of baseline neutrophil-to-lymphocyte ratio (NLR) in the curative effect and prognosis of breast cancer patients treated with neoadjuvant chemotherapy.Methods The clinical data of 304 breast cancer patients received neoadjuvant chemotherapy in Beijing Cancer Hospital during 2010 were retrospectively analyzed.The correlations of baseline NLR with clinicopathological parameters of breast cancer patients were analyzed by Wilcox or Kruskal-Wallis tests,and the correlation of NLR with pathologic complete response (pCR) after neoadjuvant chemotherapy was analyzed by the binary logistic regression.The effect of clinic pathological parameters on the disease-free survival(DFS) of the patients was evaluated by the univariable and muhivariable Cox regression models.Results The medians of NLR in pCR + patients (n =62) and pCR-patients (n =242) were 1.76 and 1.72,respectively,and there was no significant difference between themn (P ＞ 0.05).Multivariate analysis indicated that the TNM stage (P ＜ 0.05) and pCR (P ＜ 0.05) were the independent risk factors influencing the prognosis of breast cancer patients.There was no significant correlation between NLR and 5-year DFS (HR=0.842,95％CI=0.566-1.255,P＞0.05).Condusion NLR lacks the predictive value in the curative effect and prognosis of breast cancer patients treated with neoadjuvant chemotherapy,while TNM stage and pCR are the independent risk factors influencing the prognosis of breast cancer patients.

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Distribution characteristics and clinical significance of neutrophil VCS parameters in patients with zinc oxide toxicosis

Xuqian FANG

Chinese Journal of Clinical Laboratory Science.2017;35(2):111-113. doi:10.13602/j.cnki.jcls.2017.02.08

Objective To observe the clinical value of neutrophil VCS parameters in the patients with zinc oxide toxicosis.Methods The peripheral blood samples from 120 patients with zinc oxide toxicosis,50 patients with bacteria infection and 60 healthy subjects were tested using Coulter LH 750 hematology analyzer.VCS parameters,including mean neutrophil volume (MNV),neutrophil distribution width (NDW),mean neutrophil conductivity (MNC) and mean neutrophil scattering value (MNS),were examined before and after treatment.Results MNV in patients with zinc oxide toxicosis increased significantly compared with those of healthy group and bacterial-infectious group.On the contrary,MNC decreased significantly in the patients with zinc oxide toxicosis compared with that of healthy group and bacterial infection patients.There was significant statistical difference between the two groups (P ＜ 0.01).When the cut-off value is less than 144.70,the sensitivity and specificity of differential diagnosis are 90.83％,98.33％,respectively,higher than those of WBC count and other VCS parameters.The values of MNC in WBC greater than or equal to 10.0 × 109/L group and WBC less than 10.0 × 109/L group were (140.05 ±4.26) and (138.87 ±5.87) respectively.There was no significant difference between the two groups (P ＞ 0.05).The values returned to normal range after treatment for 3 days.Conclusion The MNC of neutrophil may reflect the morphologic change of neutrophils in zinc oxide toxicosis and restore its normal value after treatment,so it can be used as a marker for diagnosis and monitoring of disease.

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Establishment and verification of personalized reference interval of blood cells

Bo YUE ; Manjiao LIU ; Dahai TANG ; Jialei HE ; Mengjie JIANG ; Xiaoqiang WEI ; Danjie LI ; Shangzhen ZHANG

Chinese Journal of Clinical Laboratory Science.2017;35(2):107-110. doi:10.13602/j.cnki.jcls.2017.02.07

Objective To establish and verify the personalized reference interval of blood cells.Methods The results of blood cells from 2 089 health subjects in 2014,2015 and 2016 were collected.The ratio of the later results to the previous results was defined as the fluctuation (λ).The ratio (λ1) of the results of 2015 to the results of 2014 was calculated and λ1 was analyzed statistically to establish the fluctuation reference interval (CIλ).The ratio (λ2) of the results of 2016 to the results of 2015 was calculated.λ2 was used to verify λ2.The personalized reference interval (CIp) was established by multiplying each result of 2015 and CIλ.CIp was verified by results of 2016.The ratio of the upper and lower limits of CIp was calculated.The ratio of the upper and lower limits of the reference interval (WS/T 405) was calculated.Results The values of CIλ were as follows:WBC (0.66 to 1.53),L(0.67 to 1.51),M (0.50 to 2.00),N(0.56 to 1.78),E(0.4 to 2.51),PLT(0.76 to 1.32),RBC(0.92 to 1.12),Hb(0.92 to 1.11),Hct(0.91 to 1.12),MCV(0.95 to 1.07),MCH(0.95 to 1.05)and MCHC(0.94 to 1.06).The validation tests of CIλ and CIp showed that both CIλ and CIp were suitable for this laboratory.Compared with the reference interval of professional criteria,the ratio of the upper and lower limits of the CIp was smaller than that of traditional criteria.Conclusion CIp for this laboratory was established and verified.Compared with traditional criteria,CIp should be more personalized and highly sensitive.

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Clinical significance of the detection of serum cardiac troponin Ⅰ autoantibodies in the patients with coronary atherosclerotic heart disease

Jinhua ZHANG ; Kui ZHANG ; Hongxia WEI ; Fang QIU

Chinese Journal of Clinical Laboratory Science.2017;35(2):132-135. doi:10.13602/j.cnki.jcls.2017.02.14

Objective To observe the positive rate of serum anti-cardiac troponin Ⅰ autoantibodies (anti-cTnI) in the patients with coronary atherosclerotic heart disease(CHD) and its clinical significance.Methods A total of 200 CHD patients and 200 healthy controls from the Department of Cardiology,Drum Tower Hospital of Nanjing University Medical School during September 2015 and May 2016 were collected in the case-control study.Serum anti-cTnI was detected with an indirect ELISA method.Meanwhile,the general data,chnical biochemical parameters and coronary vascular lesion from the two groups were analyzed.Results The positive rate of anti-cTnI in CHD patients(21/200,10.5％) was significantly higher than that in healthy controls(4/200,2.0％,P ＜ 0.01).According to the detection results of serum anti-cTnI,the participants were divided into three groups,including anti-cTnI positive group with CHD,anti-cTnd negative group with CHD and anti-cTnI positive group in healthy controls.The median age and serum Glu level in the anti-cTnI positive group with CHD were significantly higher than those in the anti-cTnI positive group in healthy controls (P ＜ 0.05),while the levels of serum HDL-C and apoA I were inverse (P ＜ 0.01).The median age and Gensini score in the anti-cTnⅠ positive group with CHD were significantly higher than those in the anti-cTnI negative group with CHD,while the level of serum HDL-C was inverse (P ＜ 0.05).Conclusion Anti-cTnI may be more likely to occur in old CHD patients with severe coronary artery lesions,and the low levels of serum HDL-C and apoA Ⅰ may correlate to the production of anti-cTnⅠ.

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