Consistency evaluation of thyroid stimulating hormone results from 4 chemiluminescence assays / 临床检验杂志

Objective@#To compare the consistency of thyroid stimulating hormone (TSH) results from four chemiluminescence assays. @*Methods@#A total of 102 fresh serum samples from Peking Union Medical College Hospital during March 2018 and April 2018 were collected for precision evaluation and methodological comparison referring to CLSI EP15-A2 and EP9-A2 protocols. The levels of serum TSH were detected by Abbott i2000 (system A), Beckman DXI800 (system B), Siemens ADVIA Centaur XP (system C) and Roche e601 (system D) automatic chemiluminescence analyzers and their matching reagents, respectively. The obtained results were compared with the passing-bablok and Bland Altman methods. Taking 0.27 μIU/mL and 5.33 μIU/mL as the medical decision level, the expected bias of each detection system was compared. @*Results@#The precisions of systems A,B,C and D were 1.7%-3.3%， 2.3%- 3.9%，0.7%-2.3% and 0.6%-1.5%,respectively. The median (P 25,P 75) of TSH concentrations detected by systems A,B,C and D were 1.898 (0.518,4.809)μIU/mL, 2.819 (0.719,7.020)μIU/mL,2.502 (0.692,6.888)μIU/mL and 3.105 (0.886, 7.905)μIU/mL, respectively. The coefficients of determination (R 2 ) of regression equation were above 0.975 for 4 detection systems. The correlation coefficients (r), intercepts and slopes of 4 detection systems were 0.993 5-0.997 1, 0-0.06 and 0.59-1.15, respectively, and systems B and C had the best correlations with 1.02 of slope and 0 of intercept. The deviation plot showed that the bias% of 4 detection systems was between -48.1% and 17.3%. Among them, systems A and D had the largest bias, while systems B and C had the lowest bias. The expected bias of 4 detection systems at the medical decision level was -40.7%-37.2%. @*Conclusion@#The consistency between Beckman and Siemens TSH detection systems is good, while those of Roche and Abbott TSH detection systems are different from the other two.

Survey for variation of prevalence of hyperuricemia and its correlations with blood glucose and lipid in healthy adults receiv-ing physical examination at Peking Union Medical College Hospital from 2012 to 2017 / 临床检验杂志

Objective To investigate the changes of prevalence of hyperuricemia ( HUA) and its correlations with blood glucose and lipid in healthy adults receiving physical examination at Peking Union Medical College Hospital (PUMCH) from 2012 to 2017. Meth-ods An observational approach was adopted for the data analysis.The test results of uric acid (UA),fasting blood glucose (FBG),to-tal cholesterol (TC),triacylglycerol (TG),low density lipoprotein cholesterol (LDL-C),high density lipoprotein cholesterol (HDL-C), creatinine (Cr) and Urea of 399 089 cases (206 881 males and 192 208 females) at PUMCH from January 2012 to December 2017 were collected and statistically analyzed.Results The total prevalence of HUA was 17.4% in which the prevalence of males was signif-icantly higher than that of females (25.6% vs 8.5%,χ2＝20 234.850,P＜0.01).During the years of 2012 to 2017,the prevalence of HUA was 26.5%,24.7%,28.6%,23.9%,24.8% and 24.5% in males,and 13.8%,6.3%,7.9%,6.1%,6.2% and 6.8% in females for each year respectively.The prevalence of HUA in males aged 18 to 64 years old was significantly higher than that in the age-matched fe-males (all P＜0.05).However, the prevalence of HUA in males aged≥65 years old was similar to the age-matched females.There was no statistically significant difference of HUA prevalence between males and females aged ≥65 in 2013,2015,2016 and 2017 ( χ2＝1.792,0.017,1.440 and 0.205 respectively;all P＞0.05).The percentages of hyperlipidemia in both males and females of HUA group were higher than those of non-HUA group respectively (all P＜0.01).The percentage of hyperglycemia in males of non-HUA group was higher than that of HUA group,but the percentage of hyperglycemia in females of non-HUA group was lower than that of HUA group ( all P＜0.01).High levels of TC,TG and FBG were risk factors of HUA with increased OR values in increased concentrations of TC,TG and FBG,respectively.Conclusion During the recent 6 years, in healthy adults receiving physiced examination at PVMCH, the preva-lence of male HUA diagnosed was at overall high level,but the prevalence of female HUA was in decreasing and relatively stable trend. Hyperlipidemia and hyperglycemia should be the risk factors of HUA.

Investigated the prevalence of hypouricemia in Peking Union Medical College Hospital / 中华检验医学杂志

Objective To investigate the prevalence and possible factors of hypouricemia in Peking Union Medical College Hospital.Methods A retrospective investigation.Serum uric acid, lipids, glucose and other chemistry tests were analyzed among 83176 outpatients(Male:30795,Female:52381), 15849 inpatients(Male:7402,Female:8447)and 24081 healthy subjects(Male:11859,Female:12222)in Peking Union Medical College Hospital from December 2015 to April 2016.Grouped by gender and age, the prevalence of hypouricemiawas analyzed in all subjects and the etiology and possible risk factors of hypouricemia were explored among all patients.Results The serum uric acid of outpatients,inpatients and healthy subjects were 286(235-348)μmol/L, 282(226-348)μmol/L and 298(244-358)μmol/L, respectively.And the prevalence were 0.6%(499/83176),2.5%(390/15849)and 0.2%(39/24081), respectively.The prevalence of hypouricemia ofwomen was significantly higher than that ofmen(outpatients:0.7%vs 0.4%,P<0.001;inpatients:2.8%vs 2.1%,P=0.004;healthy subjects:0.30%vs 0.04%, P<0.001).After analyzing 507 hypouricemia patients, the top three clinical diagnoses that related with hypouricemia were kidney diseases, tumor and rheumatic diseases.Compared with the control group, the prevalence of hypouricemia in hypertriglyceridemia group and group with eGFR higher than 90 ml/(min· 1.73 m2)were lower(OR:0.33, 95% CI:0.21-0.50; OR:0.16, 95% CI:0.09-0.29), and the prevalence of hypouricemia in hyperglycemia group was higher(OR:1.62, 95% CI:1.12-2.35). Conclusion The prevalence of hypouricemia of Chinese women was higher than that of men and may be related with TG,Glu and eGFR.

The performance evaluation of four Lp-PLA2 activity assays / 中华检验医学杂志

Objective To evaluate the analytical performance of four lipoprotein associated phospholipase A2(Lp-PLA2)activity reagents on Beckman AU5800 automatic biochemical analyzer. Methods The remaining serum samples of 214 patients and 140 apparently healthy individuals were collected from March to July 2017 in Peking Union Medical College Hospital.These samples were used for method comparison and reference interval evaluation.According to the guidelines of EP15-A,EP6-A,EP-17 and EP7-P from Clinical and Laboratory Standards Institute(CLSI)standards,the precision, linearity, sensitivity and common interferences(e.g free bilirubin, conjunct bilirubin, hemoglobin and chyle)were assessed.According to EP9-A2,method comparisons of differents regents(Evermed,DiaSys,Hengxiao and Zhongyuan were labeled as A,B,C and D,respectively)were conducted and the differences were estimated at medical decision levels(328U/L,391U/L and 485U/L).Results The precision of four reagents were acceptable.The repeatability(CV%)of A to D were 0.5%-1.7%, 0.7%-3.0%, 0.9%-2.0% and 0.5%-3.3%,respectively.The reproducibility(CV%)were 0.7%-2.9%, 1.4%-3.2%, 1.3%-1.9%and 0.8%-4.1%,respectively.Both of those achievedlaboratory defined quality objective(<5%).The linearity of A to D were 44 -1 992 U/L,39 -1 798 U/L,13 -540 U/L and 75-1 717U/L,respectively.The regression coefficient R2 was between 0.997 and 1.000, and the correlation coefficient(r)was between 0.998 and 1.000.The interference of chyle were acceptable among these four reagents andmet the manufacturer′s requirementsor clinical needs.In a low level of Lp-PLA2,bilirubin had an obvious interferenceonreagent C;B and C were negatively affected when the hemoglobin was 4.5 g/L; and D was positively affected when the hemoglobin was 2.45 g/L.The regression coefficients R2 of A,C,D compared with B were between 0.978 and 0.995,and the correlation coefficients(r)were between 0.989 and 0.998. The expected differences at medical decision levels ranged from -240 U/L to 113 U/L.For A to D,the Lp-PLA2 activity results of 131(93.6%), 140(100%), 82(58.6%), and 128(91.4%)cases were analysed within the manufacturer′s claimed reference intervals.Conclusion The precision and linearity of the four Lp-PLA2 activity detection reagents used in automatic biochemical analyzer are good, but the anti-interference ability needs to be improved.

Performance verification of six enzymatic glycated albumin reagents / 中华检验医学杂志

Objective To validate the performance of six enzymatic glycated albumin reagents and evaluate their clinical application.Methods The performance of six enzymatic glycated albumin reagents(labled as A,B,C,D,E,F) from Beijing Jiuqiang Co, Beijing Lideman Co,Ningbomeikang Co, Beijing Haomai Co, Sichuan Maike Co and Asahi Kasei Co were assessed on Olympus AU5800 automatic biochemistry analyzer.According to the standard of CLSI,the precision,interference and linear correlation of these reagents were assessed.To assess the accuracy of GA% ,we used GA standard material whose value had been assigned using ID-LC/MS method provided by ReCCS.To do the method comparison and determine the consistency of assay, 50 fresh serum samples of T2DM outpatient and 80 fresh serum samples of apparently healthy people in Jan 2016 were tested using six kits.According to the EP28-A3C protocol, the reference range for GA%was validated in 122 apparently healthy individuals undertaking medical examination from January to February 2016 in PUMC.Results The precision,and the ability of anti-interference of the six reagents were good.The accuracy percentage deviation of six reagents was-19.3%-9.2%.The correlation coefficient of domestic reagents A to E and imported reagents F in the determination of GA% was 0.966-0.999, the average absolute bias was 7.0%-10.4%.The coincidence rate of A-E and F in determining abnormal GA% was between 88.5% and 96.9%.The coincidence rate was increased after switching to the reference range for preliminary clinical evaluation.Conclusion Six GA enzymatic kits used in automatic biochemical analyzer have high precision and strong anti-interference ability.Accuracy still needs to be improved.

Effects of hydrogen sulfide on impaired wound healing in ob/ob mice / 中国病理生理杂志

[ ABSTRACT] AIM:To investigate the role of hydrogen sulfide ( H2 S) on impaired wound healing in ob/ob mice and the underlying mechanism.METHODS:The ob/ob mice were randomly divided into 3 groups, including vehicle, in-sulin and NaHS for treatment.C57BL/6 mice were treated with vehicle as control.Full-thickness punch biopsy wounds were created on the mice.Firstly, H2 S concentrations in the skins and granulation tissues were measured.The mRNA ex-pression of cystathionineγ-lyase ( CSE) was detected by RT-qPCR.The protein expression of CSE and MMP-9 were deter-mined by Western blot.The neutrophil and monocyte/macrophage infiltration was analyzed by immunohistochemistry me-thod.The levels of tumor necrosis factor (TNF)-αand interleukin (IL)-6 were measured by ELISA.Collagen formation was measured by Masson staining.RESULTS:The H2 S levels in the skin and granulation were significantly decreased in ob/ob mice and increased in the NaHS-treated mice ( P to impaired wound healing in diabetes, which is alleviated by exogenous H2 S possibly through anti-inflammation.

Investigation on the Consistency of Five Measurement Systems for Serum Lipids / 现代检验医学杂志

Objective To explore achieving the consistent method of blood lipid examination by comparing the results of 5 dif-ferent blood lipid detection system commonly used in the use of refernce method to assign freach blood serum before and af-ter calibration.Methods Used the indoor quality control total variation (CV%)to evaluate the 5 blood lipid examination system of the imprecision.Referenced the United States Clinical and Laboratory Standardization Institution (CLSI)9A2 EP program,compared with 54 fresh blood serum in 5 commonly used examination system of Total Cholesterol (TC)and Tri-glyceride (TG),and then estimated the bias between the different detection systems and mean value.8 of the samples were determined by the reference method and estimate the bias of different system.The fresh frozen serum samples assigned by reference method were used to evaluate the above examination system,then compare and estimate the bias again with the same 54 fresh serum samples.Compared the variation of 54 samples in different detection system before and after calibra-tion.Results The TG imprecision of 5 examination system were between 3.76%~23.65%,the TC imprecision between 2.19%~23.43%,that mean the results were good,the r value of TG were between 0.996 7~0.999 6 and the TC were 0.956 2~0.996 7.But there were obvious differences between the results of the systems,and the biggest difference were 14.72%~34.21% in TG and 3.11%~14.57% in TC.After use the serum assignment by reference method,the variation of the systems has been significantly decreased.Conclusion Using the reference method to assign the fresh serum of different blood lipid detection system can effectively improve the consistency of the results.

Negative interference by calcium dobesilate in uricase-peroxidase coupled assays of serum uric acid / 中华检验医学杂志

Objective To assess the interference by calcium dobesilatein 7 peroxidase-baseduric acid assays and to determine its clinical significance.Methods In the in vitro experiments, uric acid in pooled serum with final concentrations of calcium dobesilate additions (0, 2, 4, 8, 16, 32, and 64μg/ml) were measured by 7 peroxidase-based assays.Percent Bias (%) was calculated relative to the drug-free specimen.In the in vivo experiments, changes in serum uric acid and calcium dobesilate concentrations were observed before and after calcium dobesilate administration ( baseline, 0 h, 1 h, 2 h, 3 h, 4 h, 6 h ) involunteers.The interference in different assays was assessed compared with LC-IDMS/MS method. Calcium dobesilate levels in 40 specimens from those taking calcium dobesilate were measured by HPLC method.Of the 40 specimens, 10 were selected to analyse the levels of uric acid by both peroxidase and UV measurement method to assess the impact in clinical status.Results In the in vitro study, concentrations of uric acid measured by 7 peroxidase-based assays were reduced by -6.3%to -21.2%compared with drug-free serum, when theconcentration of calcium dobesilate was16μg/ml.In the in vivo study, comparedto UA levels at 0 h, the biasesof serum uric acid determined by peroxidase method after calcium dobesilate administration(1 h, 2 h, 3 h, 4 h, 6 h) were of -3.33%, -6.79%, -7.49%, -6.07%, -4.09%, respectively.The observed uric acid concentrations for 8 participants measured by enzymatic assays were inhibited by -3.75% to -6.89% at 0 hour and by -16.9% to-22.22% at 2 hours relative to the concentrations measured by the LC-IDMS/MS method. Conclusions Calcium dobesilate produced a clinically significant negative interference with uric acid in all peroxidase-based uric acid assays,which may result in false evaluation of uric acid level in clinical status.Significant differences in the degree of interference were observed among the assays.

Assessment of the consistency of five standardized cystatin C measurement systems / 中华检验医学杂志

Objective To assess the consistency of four standardized cystatin C particle-enhanced turbidimetric assay (PETIA) and one particle-enhanced nephelometric immunoassay (PENIA) measurement systems Methods Performance verification test was conducted according to CLSI EP 15-A2 and EP9-A2. Fourty serum samples in comparative test were obtained from the remaining serum samples of outpatients in Peking Union Medical College Hospital in March 2013.Fourty serum samples were tested on Olympus AU5400 automatic biochemical analyzer ( four PETIA Cys C reagents:Shanghai Jingyuan Co ., Ltd, Beijing Leadman Biochemistry Co ., Ltd, Beijing Strong Biotechnologies , Maker Biotechnology in Sichuan , and labelled as A, B, C, D respectively) and PENIA N Latex Cys C measurement system on Siemens BNⅡ(labelled as E).Correlation analysis were performed among four PETIA methods one PENIA method Differences of each detection system were compared in the medical decision level 1,2,3,4 mg/L.The reference material ERM-DA471/IFCC was measured by five systems and bias ( percentage bias ) was calculate for each system.Results Results of systems A, B, C, D, E were 1.29(0.89-2.43), 1.30 (0.96-2.59), 1.22(0.90-2.44), 1.27(0.96-2.47), 1.14(0.82-2.05)mg/L.Chart shows bias among these five systems was small when Cys C concentration was less than 4mg/L.PETIA method A, B, C, D correlated with their mean value well , with the average deviation from their mean value ( percent deviation) at -0.017 -0.031 mg/L ( -3.1%-2.1%), and all were less than allowed bias from the biological variation (3.4%).The deviation of PETIA method A, B, C, D with their mean value in medical decision level at -0.176 -0.178 mg/L.Systems A, B, C, D correlated well with the result of PENIA method system E , and the mean deviation ( percent deviation ) was at 0.278 -0.326 mg/L ( 12.6%-18.5%) , and the deviation ( percent deviation ) in the medical decision level 0.055 -1.079 mg/L (5.51%-26.98%).Bias of PETIA method A, B, C, D Cys C system measuringERM -DA471/IFCC ranged from 0.22 to 0.39 mg/L ( 3.9%-7.0%) , which exceeded the allowable range of the reference material target value, and were larger than the allowable bias from biological variation (3.4%).Bias ( percent ) of PENIA method system E was -0.1 mg/L ( -1.7%) , within the allowable range of ERM-DA471/IFCC target value .Conclusions The consistency of four assesed PETIA Cys C reagents was relatively ideal, and improved markably after being traced to ERM-DA471/IFCC.Besides, the results of PETIA were higher than those of PENIA .Bias among these five systems was small when Cys C concentration was less than 4 mg/L, and the bias became larger in higher Cys C concentration.

The status of research on estimated glomerular filtration rate and its challenges / 中华检验医学杂志

Chronic kidney disease ( CKD ) is a worldwide public health problem.Glomerular filtration rate ( GFR ) is a key indicator for early diagnosis and accurate classification of CKD , how to estimate GFR accurately and conveniently has been a difficulty and hot-points.We describes the development of several major eGFR equations based on creatinine and cystatin C , analysis the impact of the different research methods on performance of equation , and discusses the issue in research and application of eGFR in China.Accordingly make the following recommendations ( 1 ) Expand the study size ( multiple centers) and participants number , then develop and validate eGFR equations based on Chinese population ;(2) Standardize the gold standardof GFR in the study and unify the analysis and evaluation methods;(3) Promote the consistency and standardization of creatinine and cystatin C which is the basis for the wide range of applications of the eGFR formula.

Correlation of ApoE gene polymorphism and serum uric acid in Ningxia Hui Autonomous Region / 中华检验医学杂志

Objective Toexplore the correlation of ApoE gene polymorphism and serum uric acid levels in Ningxia Hui Autonomous Region , China.Methods A case-control study.October 2011 to November 2011, five hundred twenty eight ( 296 male, 232 female ) apparently healthy individuals were studied.Questionnaires and physical examinations were performed by standard operation procedure.Fasting blood was collected for biochemistry testing including serum lipid parameters , uric acid concentration and creatinine levels.The multi-ARMS PCR was applied to determine ApoE genotypes ,and the relation of ApoE genotypes with serum lipid parameters and uric acid levels were analyzed.Non-normal distribution were compared using cause and inspection.Results The common six kinds of ApoE genotype can be detected.The total cholesterol ( TC) ,low density lipoprotein cholesterol ( LDL-C) and uric acid ( UA) levels in different genotype subgroups had statistical differences.The individuals with ε2/3 genotype had a significantly greater reductions in TC and LDL-C levels but increment in uric acid concentration than those withε3/3 and ε3/4 genotype (P<0.05).The effect of ApoE gene polymorphism on uric acid levels still remained significantly after adjustment for age , gender , region and other factors.Conclusion The ApoE polymorphism is associated with serum uric acid levels and individuals with ε2 allele have higher serum uric acid levels.

Validation study of chronic kidney disease epidemiology collaboration predictive equations based on serum creatinine and cystatin C in Chinese chronic kidney disease patients / 中华检验医学杂志

Objective To validate the four chronic kidney disease epidemiology collaboration (CKD-EPI) predictive equations based on serum creatinine (SCr) and cystatin C (Cys C) in Chinese CKD patients,and try to develop the GFR predictive equations for Chinese CKD patients.Methods254 CKD patients were randomly selected from four Grade ⅢA hospitals in different regions in China from September 2007 to December 2010.Clearance of dual plasma sampling 99mTc-DTPA was used to measure glomerular filtration rate (rGFR) in 254 CKD patients.The serum concentration of Cr and Cys C were measured.CKD-EPI SCr equation,Cys C equation,Cys C equation adjusted for age,sex and race,SCr/Cys C combinated equation adjusted for age,sex and race were used to estimate GRF ( labeled as eGFR1,eGFR2,eGFR3 and eGFR4,respectively).The correlation,bias and precision of eGFRs were compared with rGFR by Wilcoxon signed rank test,intraclass correlation coefficient (ICC) and Spearman correlation analysis.The deviation degree between rGFR and different eGFRs was compared via Bland-Altman graph.The accuracy within 15％,25％,30％ ( P15,P25,P30) and the staging correctness of eGFR against CKD at different stages was calculated.ResultsThe rGFR in 254 CKD participants was [ 48.07 (26.19 -92.97 )] ml · min -1·(1.73 m2) -1.The Spearman correlatiou coefficients (CC) of eGFR and rGFR varied within the range of 0.873 - 0.896 ( P =0.000 ).The intra-class CC ( ICC ) varied within the range of 0.920 - 0.942.The correlation of eGFR4 was the best.The absolute deviations of 4 eGFRs and deviation precision were eGFR4 ＜eGFR3 ＜ eGFR2 ＜ eGFR1.The 95％ confidence intervals for the regression line of 4 eGFRs shown by Bland-Altman graphs were 92.5,87.3,83.0 and 76.1 ml · min-1 · ( 1.73 m2 ) -1,respectively,with the best result of eGFR4.For P30,the correctness of 4 eGFRs were eGFR4 ＞ eGFR3 ＞ eGFR2 ＞ eGFR1,but no significant difference was found by Chi square test (x2 =6.448,P =0.092).The overall correctness rate in 4 eGFRs against CKD stages were 48.4％ -57.5％,with the highest consistency of eGFR4,but their staging correctnessratewerenotideal(Kappa values were 0.405,0.348,0.366 and 0.463,respectively).Conclusions Compared with CKD-EPI SCr equation,no advantage was found in CKD-EPI Cys C equation.The Cys C equation adjusted by age and sex shows a little advantages over CKD-EPI Cys C equation in bias,precision,correlation and accuracy.The CKD-EPI SCr/Cys C combinated equation adjusted by age,sex and race has advantage over other three equations not only in bias,precision,correlation and accuracy,but also in staging correctness.However,the validation of this equation is still not fairly ideal for Chinese CKD patients.Based on these findings,it is essential for the Chinese CKD patients to develop SCr/Cys C combined predictive equation which adjusted by age,sex or other factors.(Chin J Lab Med,2012,35:798-804)

Creatinine concentration of 2630 healthy students in primary and middle schools in Inner Mongolia / 中华检验医学杂志

ObjectiveTo analyze the serum creatinine level among apparently healthy primary and secondary school students in Inner Mongolia and explore the distribution of serum creatinine by ethnic,regional,gender and age,and establish the reference interval of serum creatinine in different gender and different age groups of primary and secondary school students.MethodsLargesample clinical epidemiological investigation was applied by two-stage clustering sampling method.Random sample of 2630 primary and secondary school students from 9 to 18-year-old was selected from four district in Inner Mongolia including Hohhot,Wulanchabu,Xilin Gol and Bayan Drow from July 2009 to June 2010.After screening outlier individual,the total of 2614 subjects were enrolled,involving 1288 male and 1326 female subjects,1584 Han and1030 Mongolian.The venous blood was collected and serum was separated.The serum creatinine concentration was measured as soon as possible.Furthermore, creatinine levels of different regions,ethnic,gender and age group were compared by analysis of variance or t-test and that of different group were compared by SNK method.Percentile was used to describe the distribution of serum Cr level of different age groups.The reference interval of serum Cr for primary and secondary school students were established by gender and age (P2.5 -P97.5 ).The curve was smoothed using age-specific percentile ( LMS )curve smoothing method.ResultsThe differences of Cr levels were statistically significant between different regions,ethnic,gender and age groups.The reference intervals of creatinine for 9 - 11,12,13 - 14,15,and 16 - 18 year-old males were 35 - 66,37 - 73,39 - 78,47 - 87 and 49 - 91 μmol/L,respectively.The intervals for 9 - 10,11 - 12,13 - 15,and 16 - 18 year-old females were 32 - 60,34 - 63,38 -73 and 40 -74 μmol/L,respectively.Conclusion The reference intervals of serum creatinine for health primary and secondary school students in the Inner Mongolia is established,which is useful for clinicians,especially pediatricians to judge and assess renal function for 9 to 18 year-old patients.( Chin J Lab Med,2012,35:805-809 )

Construction of a bicistronic expression vector containing human papillomavirus (HPV) type 6b L1 gene and its expression in mammalian cells / 中华皮肤科杂志

Objective To construct a bicistronic expression vector containing HPV type 6b L1 gene, to express the recombinant vector in mammalian cells, and to establish a cell strain stably expressing HPV6b L1 gene. Methods After endonuclease digestion and purification, the gene fragment of HPV6b L1 was cloned into the eukaryotic expression vector pIRES2-enhanced green fluorescent protein (EGFP). The identification of the recombinant was realized via endonuclease digestion and sequence analysis. Then, the recombinant plasmid pIRES2-HPV6bLl-EGFP was transfected into NIH3T3 (a mouse embryonic fibroblast cell line) cells. Subsequently, the expression of EGFP was observed by fluorescent inverted microscopy, and HPV6b L1 mRNA expression by reverse transcription (RT)-PCR. Results The recombinant plasmid pIRES2-HPV6bLl-EGFP was successfully constructed, transfected into N1H3T3 cells, and selected by G418. The expression of EGFP was seen under an inverted fluorescence microscoy. RT-PCR proved the expression of HPV6b LI mRNA in transfected cells. Conclusions The recombinant plasmid pIRES2-HPV6bLl-EGFP was successfully constructed and transfected into NIH3T3 cells. Inverted fluorescent microscopy and RT-PCR confirmed the successful expression of HPV6b L1 in NIH3T3 cells.

Impacts of different creatinine detection methods on the efficacy of different GFR estimation equations / 中华检验医学杂志

ObjectiveTo investigate the impacts of different serum creatinine detection methods,including Jaffe and enzymatic methods,on the efficacy of different GFR estimation equations in CKD patients in China.MethodsrGFR of 176 patients with CKD were determined by dual plasma sample method 99mTc-diethylenetriamine pentaacetic acid (99mTc-DTPA) plasma clearance rate.Serum creatinine was detected with four kinds of creatinine reagents from different manufacturers.Cockcroft-Gault Equation corrected for body surface area (CG/BSA),simplified Modification of Diet in Renal Disease (MDRD) Study equation,IDMS-traceable MDRD equation,CKD epidemiology collaborative research (CKD-EPI) equation and two Chinese simplified MDRD equation (project group equation 1,2) were applied to calculate estimated GFR (eGFR)respectively.eGFRwerecomparedwithrGFRforthecorrelation, deviation, precisionand30％ accuracy.ResultsThe mean rGFR of 176 patients with CKD,was [ 40.70 ( 19.41 -84.35 ) ] ml · min- 1 ·( 1.73 m2 ) -1.For all GFR estimation equations,there were significant differences in eGFR results between enzymatic method and Jaffe method,when analyzed by the Wilcoxon signed-rank test.eGFR results assessed by two enzymatic creatinine detection systems showed no significant difference,while eGFR results analyzed by two Jaffe detection system were significantly different.The intraclass correlation coefficient (ICC) of eGFR and rGFR ranged from 0.879 to 0.923 by Jaffe method,while from 0.925 to 0.946 by enzymatic creatinine method.ICC and Pearson correlation analysis revealed a significant correlation between eGFR and rGFR,and the correlation was better when using enzymatic method.Bland-Altman plots indicated that large deviation occurred in the high value area of GFR using various equations.However,deviation with the enzymatic creatinine method was smaller than that with the Jaffe method. When rGFR ≥ 60 ml · min- 1 ·(1.73 m2) -1,the 30％ accuracy of eGFR using enzymatic creatinine method for all six equations was between 68.3％ and 90.0％,while it was between 41％ and 75％ when using Jaffe method. The 30％ accuracy of eGFR using enzymatic creatinine method was significantly higher than that using picric acid method for these equations except for the project group equation 1.When rGFR ＜60 ml · min -1 · ( 1.73 m2 ) -1,the 30％accuracy of eGFR using both methods was between 39.7％ -49.1％,40.5％ -52.6％respectively,and the difference of data showed no statistical significance.For the same equation,there was a significant differernce in 30％ accuracy of eGFR between two enzymatic creatinine detection systems,while there was no significant differernce between two Jaffe creatinine detection systems.ConclusionsA significant difference was demonstrated in the same GFR evaluation equation using two different creatinine detection methods (Jaffe method and enzymatic method).The correlation between rGFR and eGFR,the degree of deviation,and accuracy of eGFR results assessed by enzymatic creatinine method were better than those by Jaffe method.The eGFR results assessed by different enzymatic detection systems revealed no significant difference.

Analytical performance of three cysteine proteinase inhibitor c reagents applied in the automatic biochemistry analyzer / 中华检验医学杂志

Objective To validate the analytical performance of three Cys C reagents with particle-enhanced turbidimetric immunoassay(PETIA) method used on the automatic biochemistry analyzer for preliminary clinical application.Methods The performance of three Cys C reagents (labeled as A, B, C) with PETIA method from Shanghai Jing Yuan Co., Beijing Leadman Co. and Beijing Jiuqiang Co. on OlympusAU2700 automatic biochemistry analyzer were assessed.According to the standard of CLSI EP6-A, EP15-A and EP7-P, the precision, linearity range, disturbance (bilirubin, hemoglobin, chyle) were assessed, and compared with those of Cys C reagent based on particle-enhanced nephelometric immunoassay(PENIA) from Dade Behring Co.. The reference ranges for Cys C in serum of 120 healthy individual were evaluated.Results The within-run CVs of the three reagents (A, B and C) were 3.08%-3.2%, 2.3%-4.15% and 1.38%-1.53% respectively.The total CV in A, B and C were 3.29%-3.44%, 2.65%-5.18% and 1.67%-1.69% respectively, lower than the stated.Limits of quantitative determination (LOQ) of the three reagents were 0.41, 0.23 and 0.07 mg/L, basically meeting the testing requirement.The linearity range was 0.22-7.26 mg/L(r=0.996), 0.20-7.72 mg/L(r=0.999)and 0.20-7.62 mg/L(r=0.997)in the three reagents, which demonstrated a sound linear correlation. For interference tests, no remarkable interference (<±10%) of reagent C was detected when bilirubin≤684 μmol/L, hemoglobin≤9.7 g/L and Chyle turbidity≤6 200 FTU; and no significant interference of reagent B was found when bilirubin≤684 μmol/L, hemoglobin≤6.79 g/L and Chyle turbidity≤6 200 FTU; when bilirubin≤684 μmol/L, hemoglobin≤4.85 g/L and Chyle turbidity≤1 240 FTU reagent A was not interfered significantly. The comparison afte and before the high-speed centrifugation reveals that the average percentage of bias for reagents A, B, C measured Cys C in chylous serum samples of patients was -8.31%, 1.52%, 1.32%, respectively.In method comparison tests, the regression equations of the three reagents compared with Dade Behring PENIA Cys C reagent were as follows:Y=0.787X+0.492 (R2=0.976), Y=1.098X+0.137 (R2=0.982) and Y=1.037X+0.249 (R2=0.996), respectively. Agreement rates of the high Cys C in reagent A, B, C and Dade Behring Cys C reagent were 80% (Kappa=0.615,P=0.000), 100% (Kappa=1.000,P=0.000), 91.2% (Kappa=0.824,P=0.000); While for reference range of preliminary clinical assessment, diagnosis coincidence rate of reagent A increased to 98.8% (Kappa=0.974,P=0.000). Conclusions When used in automatic biochemical analyzer, the three Cys C reagent with PETIA showed high precision,sensitivity, and sound correlation with Dade Behring PENIA reagents.The three reagents are all able to meet clinical test requirements, nevertheless, anti-interference capability were diffierent and the reference range should be further validated.

Application of Cystatin C-based formulas for prediction of glomerular filtration rate / 中华检验医学杂志

Objective To investigate the applicability of Cys C-based formulas for prediction of GFR in Chinese patients with CKD.Methods A total of 176 adult patients with CKD including 90 males and 86 females collected from 4 hospitals located in different geographic regions of China (Beijing,Shanghai,Dalian and Changsha) were enrolled in this study from September 2007 to July 2009.The rGFR was measured using 99mTc-DTPA clearance rate two-sample method.Cystantic C was measured by PETIA and PENIA respectively.The results of eGFR in the Larsson formula,Grubb formula,Hoek formula,Filler formula,Stevens formula and Hojs formula were compared with the rGFR to evaluate the calculation coherence,bias,precision,accuracy and the performance of correct phasing of the formulas.Results The mean 99mTc-DTPA clearance was [40.70 ( 19.09 - 86.49)] ml · min-1 · ( 1.73m2 ) -1.Significant difference was witnessed in the evaluation of GFR estimation formulas calculated by PETIA and PENIA (P ＜0.01).ICC and Spearman correlation analysis revealed a significant correlation between eGFR and rGFR.The ICCs of eGFR and rGFR ranged from 0.874 to 0.938.Compared with rGFR,the 30％ accuracy of all the eight evaluation formulas using PETIA and PENIA method were lower than 60％.The percentages of correct phasing in all the 5 stages of CKD were not ideal.With these formulas,percentages of correct phasing from CKD stage 2 to CKD stage 4 were lower than 65％.The eGFRs were underestimated by formulas evaluated by PENIA in CKD stage 1.All the eGFRs were overestimated remarkably by all equations in CKD stage 5.Conclusions None of the eight Cys C based formulas are ideal for estimation of GFR in Chinese CKD patients,and they can not be applied to Chinese patients directly.For this patient population,further studies will be needed to develop a more accurate Cys C-based GFR estimation formula that includes ethnicity,age and other factors.

Expressions of survivin, COX-2, VEGF and their correlations with angiogenesis in condyloma acuminatum / 中华皮肤科杂志

Objective To investigate the expressions of survivin, cyclooxyenase-2 (COX-2) and vascular endothelial growth factor (VEGF) and their relationship with angiogenesis in condyloma acuminatum (CA) tissues. Methods Immunohistochemistry using PowerVision staining kit was performed to detect the expression of survivin, COX-2 and VEGF protein in 60 CA tissue samples from patients and 21 normal skin samples from the foreskin of human controls. At the same time, the microvessel density was determined in CA tissues by staining blood vessel endothelium with anti-CD105 monoclonal antibody. Results The positivity rate of survivin and COX-2 expression was 56.67% and 63.33%, in CA tissues, 9.52% and 0 in normal skin tissues, respectively. There was a significant difference between the two groups of tissue samples in the positivity rate and intensity of survivin and COX-2 expression (all P < 0.05). VEGF was expressed in all of the CA tissues and normal skin tissues, while the intensity of VEGF expression was statistically different between the two groups of tissue samples (P < 0.05). The MVD was 16.38 ± 5.46 and 0.62 ± 0.44 in CA tissues and normal skin tissues, respectively (P < 0.05). There was a significant positive correlation between the expressions of survivin, COX-2 and VEGF, as well as between MVD and the expressions of survivin and COX-2 in CA tissues. Conclusion The expression levels of survivin, COX-2 and VEGF are significantly higher in CA tissues than in normal skin tissues.

Immunoblot analysis of hepatitis C antibody-weak positive samples detected by chemiluminescent assay / 中华检验医学杂志

ted by the ECI analyzer.

Study on Methylation of p15(INK4B) Gene in Acute Myeloid Leukemia and Chronic Myeloid Leukemia / 中国实验血液学杂志

In order to explore the role of p15(INK4B) gene with highly methylated CpG island in the pathogenesis of leukemia, the expression levels of p15(INK4B) gene was detected in patients with AML and CML. Methylation-specific PCR (MSP) assay was employed in the experiments. The methylation incidence was 83.9% (26/31) in AML and 0% (0/28) in CML. The results showed that methylation of p15(INK4B) gene was the one of the major ways for inactivation of the gene, and the methylation could be appeared in clinical development of the disease and patients condition worsened. Methylation of p15(INK4B) did not occur and its function probably is normal in CML.