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Objective:To establish a matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) method for the direct detection of serum M protein without antibody enrichment, and to assess its detection performance.Methods:Method establishment. A total of 712 waste serum samples were collected from patients who applied for the M protein identification test in Beijing Chaoyang Hospital affiliated to Capital Medical University. The immunoglobulin light chain was obtained by reduction of IgG and IgA by TCEP, and the detection method was preliminarily determined. The waste serum samples from 20 healthy people were collected to determine the range of mass-to-charge ratios of κ and λ light chain ions. 8 parallel tubes and 8 batches were set up for intra-and inter-batch reproducibility evaluation. 10-fold, 100-fold and 200-fold diluted M protein from 23 positive samples were detected by established MALDI-TOF MS method, and its sensitivity was evaluated. 3 methods of IFE, SPE and MALDI-TOF MS were used to detect M protein simultaneously, and the coincidence rate between MALDI-TOF MS and IFE and SPE was calculated.Results:The repeatability within and between batches was 100%, respectively. The original, 10-, 100-and 200-fold dilutions of 23 M protein-positive samples were determined, and the detection limit of MALDI-TOF MS for M protein was 0.06-0.18 g/L. IFE as the gold standard, the overall coincidence rates of SPE and MALDI-TOF MS were 85.9% and 92.3%, respectively, and the positive coincidence rates of SPE and MALDI-TOF MS were 72.8% and 99.7%, respectively, of the 712 samples. Among the different types of M-proteins, MALDI-TOF-MS agreed 100% with IFE M-protein results for IgA, IgD, IgM, free light chain type and biclonal group, while the agreements of SPE for IgM, IgA and free light chain samples were only 66.7%, 58% and 19.5%, respectively. One positive sample in the IgG group was not detected by MALDI-TOF MS. 23 M-proteins positive samples were diluted by original, 10, 100 and 200 times to access the sensitivity of MALDI-TOF MS method. The coincidence rate of MALDI-TOF MS was 100% and IFE was 96% at 10-fold dilution. The coincidence rate of IFE was 28% and 23% of MALDI-TOF MS at 100-fold and 200-fold dilution, respectively.Conclusions:A MALDI-TOF MS method for the detection of serum M-proteins was successfully established. This method has the advantages of high detection throughput, fast speed, good sensitivity, specificity and coincidence rate.
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Objective:To prepare the control materials of point-of-care(POC) glucose testing and evaluate their homogeneity, stability and matrix effects.Methods:The high, medium and low concentration control materials were prepared from patient leftover whole blood, which was centrifuged, fixed, washed, filtered, and aliquoted. The homogeneity and stability of the control materials were evaluated according to CNAS (China National Accreditation Service for Conformity Assessment, CNAS) GL29:2010"Reference materials-General and statistical principles for certification". The control materials were used to evaluate the matrix effects in POC glucose detection systems by Deming regression, according to the Clinical and Laboratory Standards Institute (CLSI) EP14-A3. Meanwhile, these control materials were used as the internal quality control, and their coefficients of variation ( CV) were calculated. One-way ANOVA and t-Test were used to analyze the results. Results:The homemade materials at three concentrations showed good homogeneity[ F< F0.05(9, 20)]. When the control materials were stored at 2-8 ℃, the stable phases for the opened and closed bottles were 10 days and 15 days, respectively, and there was no statistically significant difference between the results of the first day( P>0.05). The control materials at three concentrations also showed good applicability and there were no matrix effects in 10 POC glucose systems. When the control materials were detected in the internal quality control, the CVs of the high, medium and low concentrations were 0.63%, 0.66% and 1.65%, respectively, which were all below 7.5%. Conclusions:The homemade human control materials of POC glucose testing showed good homogeneity, stability and applicability. They met the requirements of quality control in hospital settings, which provided a good application prospect of the quality management of POC glucose testing.
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Objective:To establish the sex-, age-and season-specific (month) reference intervals (RI) for thyroid stimulating hormone (TSH) measurement by big data and indirect method in adults.Methods:TSH data of anonymous patients were collected from Beijing Chaoyang Hospital Affiliated to Capital Medical University in 2016, the data were selected and outliers were removed. Indirect methods (Hoffmann method and Bhattacharya method) were used to calculate TSH reference intervals of whole population, different genders, ages and seasons (months). TSH RI from two indirect methods of total population, selected population, physical examination population was compared with RI from reagent instruction according to reference change value ( RCV) based on biological variability. Results:A total of 61 599 records were obtained from 90 699 records including 18 776 males and 42 823 females. The TSH RI were obtained by Hoffmann method: the whole population, 0.59-5.59 μIU/ml (1 μIU/ml=1 mIU/L), male, 0.53-5.16 μIU/ml, female, 0.59-6.11 μIU/ml. The upper limits of TSH RI were higher with age and in winter (January): 18-30 years old, 0.62-5.57 μIU/ml, 71-80 years old, 0.49-6.45 μIU/ml; January, 0.59-6.40 μIU/ml, August, 0.60-5.56 μIU/ml; The RI of TSH by Bhattacharya method: the whole population, 0.58-5.80 μIU/ml, male, 0.55-5.02 μIU/ml, female, 0.62-6.21 μIU/ml. The upper limits of TSH RI were also higher with age and in winter (January): 18-30 years old, 0.65-5.67 μIU/ml, 71-80 years old, 0.46-5.99 μIU/ml, January: 0.61-6.52 μIU/ml, August: 0.61-5.69 μIU/ml. Compared to RI from reagent instruction, the differences of TSH RI from two indirect methods of total population, selected population, physical examination population were acceptable.Conclusions:TSH RI was established by indirect method. With the increase of age and winter, the upper limit of TSH reference interval tends to increase.
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Objective To prepare the trueness verification materials of C-reactive protein (CRP) and evaluate its homogeneity, stability and commutability. Methods The high and low CRP concentrations trueness verification materials were from patient leftover sera which were pooled, mixed thoroughly, filtered and aliquoted. The homogeneity, stability and commutability of these materials were evaluated according to CNAS(China National Accreditation Service for Conformity Assessment, CNAS)-GL29:2010 "Reference materials-General and statistical principles for certification (ISO Guide35:2006)"and the Clinical and Laboratory Standards Institute (CLSI) EP30A. The trueness verification materials were used to evaluate the commutability in 10 clinical CRP detection systems, using forty-five patients' leftover sera with different CRP concentration evaluated by Deming regression in EP30A of CLSI. Meanwhile, the commutability of dilution series of ERM DA-474/IFCC were evaluated using the same method. Results A total of two CRP concentration level trueness verification materials were prepared, with high and low concentration levels of 754 and 743 vials, 1 ml each, respectively. The preparation showed good homogeneity (F<F0.05(14,30);On the condition of room temperature, 2-8 ℃ and -80 ℃, these materials were stable for 7 days and 44 months respectively, the slope of the linear equation of | b1 | less than t0.95,n-2 · s(b1), there was no statistically significant difference between the slope and zero, the stability is satisfied. The materials and the dilution series of ERM-DA 474/IFCC also showed good commutability among patient sera in 10 systems. Conclusions The trueness verification materials of C-reactive protein (CRP) showed good homogeneity, stability and commutability. The dilution series ERM DA-474/IFCC also have good commutability. These provided experimental support for the value transfer and application of the trueness verification materials .
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Objective To explore the CRP harmonization by calibration using commutable trueness verification materials. Methods High and low level of CRP concentrations trueness verification materials(H and L) were prepared by Beijing center for clinical laboratories. Thesetrueness verification materials were diluted to 5 calibration points(5L, 4L+1H, 3L+2H, 1L+4H, 5H) by weighing method, respectively. These 5 points were used to calibrate four different brands of CRP detection system (Diasys, Leadman, Siemens and Roche) instead of the original procedure. Sera from 21 patients and the international standard ERM DA-474/IFCC were used to compare harmonization and trueness after calibration. Each sample above was measured twice. Results After calibration, the median of CV was reduced from 19.33％ to 2.92％ among 21 patient samples, less than the optimal CV based on biological variability (CV=10.6％). Compared with Desai, the slopes were closer to 1 from 0.90-1.09 to 0.93-0.96 after calibration. Meanwhile, if ERM-DA474/IFCC was used as the trueness verification materials, the absolute bias wasreduced from 3.08-11.07 mg/L to 0.52-2.97 mg/L which was close to theuncertainty of itself (2.5 mg/L). Conclusions Afterthe calibration which contained five linear concentration points of CRP trueness verification materials by weighing method, both harmonization and trueness of CRP were improved.
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Objective To value C-reactive protein ( CRP ) trueness verification materials and to perform the CRP trueness verification program in Beijing .Methods The CRP value of trueness verification materials were assigned by the international reference material ERM DA-474/IFCC, using 10 clinical routine detection systems at departments of clinical laboratory of Beijing Chaoyang and Luhe Hospital Affiliated to Capital Medical University .The calibration curves with 4 ERM DA-474/IFCC dilutions were established and used for value transfer for trueness verification materials of two levels .The uncertainty was also assessed during the process.Then, the trueness verification was performed in the EQA at Beijing Center for Clinical Laboratories ( BCCL ) among 42 clinical laboratories.The samples were distributed according to BCCL standard operating procedure .The Microsoft Excel 2007 and SPSS 17.0 were used to process the results and the function of efficiency ( En) was calculated to verify the difference between the value and the overall mean of all participating laboratories .Results The values and uncertainties of two trueness verification materials of CRP were (109.9 ±9.4) mg/L and (27.1 ±2.4) mg/L respectively.The results of trial application of two level trueness verification materials in the EQA at Beijing Center for Clinical Laboratories (BCCL) were satisfied.There were no significant difference between the transfer values from our study and the values from means of all laboratories in Beijing .The function of efficiency ( En ) was less than 1.Conclusions The valueswhich were established by using multiple detection platforms for CRP trueness verification materialswere accurate and the uncertainties were small .This method is a preferably method for CRP value assignment because there was no suitable reference method for CRP measurement till now .Thematerialswere suitable for the trueness verification program for clinical laboratories in Beijing .
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Objective:To explore the adjustment factors to the migration and functional activity on macrophages in the uterine of inflammatory mice induced by LPS.Methods:150 Kunming female mouse were divided into control group (group A),LPS model group (group B),MCP-1 blocking-up group (group C),the mice uterines were extracted separately at hour 1,3,6,12,24.The number of CD14+ macrophages and the expression of CD14 macrophages were detected by Immunohistochemistry,ELISA detects the expression of TNF-α and MCP-1.Results:①Compared with group A,the number of CD14+ macrophages and the expression of CD14 in endometrium,myometrium,perimrtrium of group B were highly significantly increased (P<0.01) at every time points,the endometrium,myometrium of group C were closely to normal level at 1,3,6 h;compared with group B,the number of CD14+ macrophages and the expression of CD14 were highly significantly decreased(P<0.01)at 1,3,6 h of perimrtrium of group C and every time points of endometrium,myometrium of group C.②Compared with group A,the content of TNF-α and MCP-1 were highly significantly increased (P<0.01) at every time points of group B and 12,24 h of group C;compared with group B,the content of TNF-α and MCP-1 of group C were highly significantly decreased(P<0.01) at every time points.Conclusion:The migration of macrophages and the expression of CD14 and TNF-α in the uterine of inflammatory mice induced by LPS were regulated by MCP-1.
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Objective To investigate the protective effects of combination alprostadil and dexmedetomidine preconditioning treatment on skeletal muscle ischemia reperfusion and secondary lung injuries.Methods Sixty patients aged less than 70 years old undergoing unilateral lower extremity surgery were randomly assigned into four groups:control group(group A),dexmedetomidine group(group B),alprostadil group(group C)and combined treatment group(group D) with 15 cases in each.In group C and D,10 μg alprostadil was given from vein at 15 min before tourniquet inflation,in group B and D 1 μg/kg dexmedetomidine was given from vein at 10 min before tourniquet inflation,the equal volume of normal saline was infused in control group.The blood samples were drawn from vein and artery for blood gas analysises and determination of malondialdehyde(MDA) and human pulmonary surfactant specific protein D (SP D) concentrations at the time before oxygen inhalation(T1),2 h after tourniquet deflation(T2),6 h after tourniquet deflation(T3) respectively.Results In group B,C and D,the alveolar arterial oxygen pressure difference(PA-a DO2)and respiratory index(R1) were significantly lower than those in group A at T2(P＜0.05).PA-aDO2 and R1 were significantly higher at T3 than those at T1 in every groups(P＜0.05);In group B,C and D,MDA and SP D were significantly lower than those in group A at T2 and T3(P＜0.05).In group D,MDA and SP D were significantly lower than those in group B and group C at T3(P＜0.05).Conclusion Alprostadil and dexmedetomidine are infused from vein preconditioning can attenuate the damages of skeletal muscle ischemia reperfusion and secondary lung injuries.The combination of alprostadil with dexmcdetomidine can produce stronger effects.
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Objective To investigate the internal quality control ( IQC ) on clinical chemistry , clinical immunology and clinical hematology in mutual recognition laboratories in medical institutions in Beijing.Methods By means of questionnaire survey and on -site investigation, fresh frozen serum and whole blood samples with assigned values by reference method were measured to investigate the status of IQC on clinical chemistry , clinical immunology and clinical hematology in 142 mutual recognition laboratories in medical institutions of Beijing,and results were analyzed.Results 142 copies of questionnaireson clinical chemistry, clinical immunology and clinical hematology were send out and 120, 97, and 101 laboratories returned the questionnaires respectively .The information feedback rate was 84.5%, 68.3% and 71.1%respectively .All the questionnaires were effective .Questionnaires survey results showed that more than 50%laboratories set up quality control goals and the most of the goals were probability for error detection ( Ped) 95%, probability for false rejection(Pfr)5%;About 70% laboratories usecd the same quality control plan for different tests ;The most frequently used quality control rules are 12s/13s/22s.On-site investigation showed that ,take the results of clinical chemistry for example , based on the desirable biological variation and WS/T 403 -2012 , most of the tests can't meet the quality control goalsunder the existing quality controlcondition.Conclusion Clinical laboratories should consider their actual situations , assess their own qualitylevels that they can reach , set reasonable quality standards for themselves , and make appropriateindividualized quality control plan.
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Objective To prepare the frozen human serum pools for AFP reference materials through quantity value transmission from several analysis systems . Methods Method establishment. According to the requirement of preparation for the national standard materials , the studies of homogeneity and stability of frozen human serum pools for AFP reference materials were carried out .The commutability of prepared AFP reference materials and WHO AFP 72/225 reference material were assessed .By use of four automated chemiluminescence analysis systems , the prepared reference materials and different dilutions of WHO reference material 72/225 were tested in the same run.The values of prepared AFP reference materials were assigned by comparing these results and the uncertainty was evaluated .Results The three levels of AFP reference materials were tested to be homogeneous .The long term stability had been observed at -80℃for 14 months.The different dilutions of WHO AFP 72/225 reference material and three levels of prepared reference materials were commutable with patient serum samples among the four analysis systems .Through multiple system quantity value transmission and total uncertainty evaluation , the assigned values ( IU/ml) of three levels of AFP reference materials were 23.0 ±3.0, 93.2 ±7.2, ( 26.1 ±2.4 ) ×102 respectively.Conclusions The three levels of AFP reference materials were homogeneous and stable in accordance with requirement of preparation for national standard materials .The assigned values were reliable.The materials showed accepted commutability across 4 analytical systems.These materials had been approved as the national certified reference materials .These reference materials could be applied for the calibration or calibration verification of clinical analytical systems and the external quality assessment schemes for clinical laboratories.
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Objective To evaluate the commutability of certified reference material IRMM ERM-DA 471/IFCC for cystatin C measurement among 8 methods.Methods 46 individual samples used for the commutability study were residual serum samples collected from clinical laboratories.The individual samples interspersedwith pooled sera and the certified reference material IRMM ERM-DA 471/IFCC and the whole set of samples was divided into 2 subsets for measurements in 2 days.The measurements were performed by8 different methods o.The samples were measured in duplicate order.Calibration was performed every day. Passing-Bablok regression was performed to compare the slopes and intercepts of mean values of the serum samples derived from different methods. Pearson correlation cofficient was also calculated. Deming regeression and 95%confidence intervals were calculated to evaluate the statistics commutability of ERM-DA 471/IFCC.The minimal specification of bias derived from biological variations was calculated to evaluate the clinical commutability.Results The within-laboratory CVs of pool sera ranged from 0.5% to 4.0%.The Passing-Bablok slope ranged from 0.765 to 1.311 and intercepts ranged from -0.04 to 0.241.The determination coefficient of Pearson regression ranged from 0.988 to 0.999.Deming regeression and 95%confidence intervals demonstrated commutability of ERM-DA 471/IFCC in 4/28 (14.3%) methods pairs. The minimal specfication bias ( 5.12%) demonstrated commutability of ERM-DA 471/IFCC in15/28 (53.6%) methods pairs.Conclusions The ERM-DA 471/IFCC domonstrated poor commutability between some methods pairs. The commutability of ERM-DA 471/IFCC should be evaluated before used as calibrators.
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Objective:To explore the immunosuppression effects and tocolysis of fructus polygoni orientalis on abortion mice induced by LPS.Methods:Mice of Kunming (55 mice) were divided into control group (group A ,10 mice) and experimental group. Experimental group were divided into group B ( intraperitoneal injection LPS ) , group C ( ingestion fructus polygoni orientalis ) , and group D (ingestion fructus polygoni orientalis and intraperitoneal injection LPS ),each group of 15 mice.Then the pregnancy results were observed ,the positive of α-NAE+and the varity of CD 14+and CD204+macrophages ,TNF-αin the uteri were identified by enzyme-histochemistry ,immunohistochemistry and ELISA.Results:The abortion rate and the embryo resorbing rate were all 100%( P<0.01 ) in group B.But there was the decreased abortion rate of 13.33% in group D .The embryo resorbing rate decreased to 10.39%.The number and positive cell area of α-NAE+and CD14+macrophages in the uteri of gestation mice of group B was greatly increased comparing with group A ( P<0.01 ) .These effects outside of myometrium of group D were remarkably increased comparing with group A (P<0.01),but there was no distinct difference in the inside of myometrium and function layer.The number and positive cell area of CD204+macrophages in group C and D was greatly increased comparing with group A and B ( P<0.01 ) .The TNF-αcontents in the uteri of mice in group B were greatly increased comparing with group A (P<0.01),but the positive cell area of CD14,CD204 were close to normal levels in group D.Conclusion:The effect of miscarriage induced by LPS is antagonized by fructus polygoni orientalis through inhibiting the phenotype ,activity and function characteristics of macrophages in the uteri of gravidity mice.
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0.05). It was a non-specific reaction to the rivanol. Conclutions The results indicate that cordocentesis is saft and reliable in clinical practice.