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OBJECTIVE@#To analyze the hematological phenotypes of Hb J-Bangkok and concomitant thalassemia.@*METHODS@#In total 72 397 samples were screened by using capillary electrophoresis. Samples with Hb J-Bangkok were identified by DNA sequencing and analysis of red blood cell parameters. Gap-PCR and PCR-reverse dot blotting (PCR-RDB) were used for analyzing the thalassemia genes.@*RESULTS@#Thirty one cases of Hb J-Bangkok were identified, all of which were heterozygotes. The hematological phenotype index (Hb, mean corpuscular volume, mean corpuscular hemoglobin, Hb J-Bangkok, Hb A@*CONCLUSION@#Hb J-Bangkok heterozygotes have normal hematological phenotypes, though they may show different hematological characteristics when concomitant with different types of thalassemia, for which genetic counseling should be provided accordingly.
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Female , Humans , Male , Hemoglobins, Abnormal/genetics , Heterozygote , Phenotype , Thailand , beta-Thalassemia/geneticsABSTRACT
Due to the characteristics such as high capture, high recovery and precise control with fluid, the microfluidic chip has attracted much attention in the research field of circulating tumor cells (CTCs). The developed microfluidic system mainly included three types based on the captured principles such as biological affinity tag microfluidic chip, free label microfluidic chip and rely on biological affinity with the physical properties of integrated microfluidic chip.
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Objective:To compare the consistency and detection capability of seven 2019-nCoV nucleic acid detection kits, and provide reference for detection method selection of clinical laboratory and diagnosis of new coronavirus pneumonia.Methods:Two batches of pharyngeal swab samples were collected from tenpatients with confirmed infection of 2019-nCoV and 10 suspected patients with negative 2019-nCoV test results during January 29 to February 5, 2020 in Shenzhen Luohu People′s Hospital. Seven kinds of kits were labeled as ato g and used for nucleic acid detection respectively to evaluate the consistency of the test results of the clinical samples. A 2019-nCoV positive specimen was selected and diluted to 5-concentration gradient plates (Level-1 to 5) with RNase-free water. The positive detection rate and intra-batch repeatability of different brands of kits were compared.Results:The negative and positive coincidence rates of twenty clinical samples tested by six kinds of kits were 100%, and the positive and negative coincidence rate was 8/10 and 10/10 for the other kit, respectively. The results of intra-batch repeatability showed the CVs of viral loads tested by these seven kits were all less than 5%. In the concentration range of Level-1 to 3, the detection capability for open reading frame (ORF)1ab gene of Kit b,d and f was lower than Kit a,c,e and g, and the detection capability of kit e and g was the highest (14/15). The detection capability for N gene of Kit a (15/15) was higher than the other 5 kits. The comprehensive analysis of the detection capability for ORF1ab and N gene showedthat Kit d had the lowest detection capability (ORF1ab:40%,N:53%), and there was no significant difference in the detection capability of Kit a, b, c, e, and f.Conclusions:There was no significant difference in the accuracy and repeatability of the seven kits for positive samples with high viral loads, and the detection performance was good; but some kits had poor detection capability for weak positive samples. It is suggested that the weak positive samples should be rechecked by at least two manufacturers′ kits to ensure the accuracy of the results.
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Objective@#To identify a α-globin gene mutation-IVS-Ⅱ-55 (T→A) and analyze hematological characteristics of IVS-Ⅱ-55 (T→G) carriers. @*Methods@#The peripheral blood samples were collected from the members of five family and three sporadic IVS-Ⅱ-55(T→G) carriers for the analysis of RBC parameters and hemoglobin electrophoresis. Gap-PCR, PCR-RDB (reverse dot blot) and DNA sequencing were carried out for the identification of gene deletion and mutation of α-globin and β-globin. @*Results@#The results of RBC parameters of five infant probands which presented with microcytic hypochromic anemia were below the normal reference interval. One of the adult carriers of IVS-Ⅱ-55 (T→G) heterozygote alone presented with microcytic hypochromic anemia, and the others showed normal RBC parameters. The hematological phenotype index (MCV, MCH and HbA 2 ) of one adult carrying a compound heterozygote for IVS-Ⅱ-55 (T→G) and βCD27-28M/N were 65.0 fL, 20.3 pg and 5.8% respectively. The hematological phenotype index (MCV, MCH, HbA 2 and HbF) of one adult carrying a compound heterozygote for IVS-Ⅱ-55 (T→G) and SEA-HPFH were 81.9 fL, 26.5 pg, 3.0% and 29.0% respectively. The HbA 2 levels of all carriers of IVS-Ⅱ-55 (T→G) heterozygote alone were in normal range. No abnormal hemoglobin band was detectable on hemoglobin electrophoresis for all the carries. @*Conclusion@#The carriers of IVS-Ⅱ-55(T→G) heterozygote alone were asymptomatic. The phenotype of compound heterozygote for β-thalassemia was similar to that of β-thalassemia alone.
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Objective To improve the efficiency of result reporting and ensure the accuracy of the results by establishing autoverification system in Clinical Chemistry and Immunology Laboratory.Methods The study followed the requirements of the Clinical Laboratory Standards Institute(CLSI)AUTO-10A and ISO 15189:2012.In addition,seven categories of verification rules were encoded using the autoverification function of the CentraLink?Data Management System on the Aptio?Automation platform.These rules included Clinical Diagnostic Standard(CS), Sample Status(SS), Quality Control Severity(QS), Instrument Error Flags Severity(IS), Normal Severity(NS), Delta Check Severity(DS), and Logical Assessment Standard(LS).Various modules of Aptio Automation,laboratory information system(LIS)and hospital information system(HIS)were integrated using the CentraLink system to establish the autoverification system.Results The autoverification system was set up and tested from August 2015 to April 2016.In total, the system ran 4 496 425 tests on 366 180 chemistry specimens.The overall autoverification rate for tests performed increased from 53.4% to 87.0%.Glucose had the highest rate (98.3%)while CKMB had the lowest rate(63.6%).Average TAT for result verification decreased by 97.7%,from 46.3 minutes to 3.7 minutes.The system ran 410,040 tests on 160 119 chemiluminescence specimens.The autoverification rate for tests performed increased from 40.2%to 89%.C-P had the highest rate(98.4%)while A-TPO had the lowest rate(58.7%).Average TAT for result verification decreased by 77.4%,from 14.6 minutes to 3.3 minutes.From May 2016 to January 2017(when autoverification was employed),compared with the same period in 2014(when manual verification was employed),the following changes were observed with no increase in staff capacity:a)Volume of routine chemistry tests increased by 46.4%,and median TAT for tests decreased by 41.9%, from 118 minutes to 83 minutes; b)Volume of chemiluminescence tests increased by 24.5%and median median TAT for tests decreased by 52.4%, from 131 minutes to 86 minutes;c)Median TAT for critical values decreased by 50.5%; d)Rates of tests that did not go through autoverification were 88.2% for NS,6.05% for SS, 2.40% for DS,2.00% for LS, 0.97%for IS,and 0.43% for CS; e)Rates of abnormal specimen status identified by Aptio Automation were 7.13‰for jaundice,5.39‰ for blood lipids,2.20‰ for hemolysis,0.17‰ for barcode error, and 0.15‰ for insufficiency;f)Error rate decreased to 0.00%;and g)staff satisfaction increased from 85%to 100%.Conclusion Autoverification of results by using the CentraLink Data Management System can achieve quality control over the entire process of clinical laboratory testing, ensure accuracy of test results, improve work efficiency, decrease TAT, minimize the error rate, avoid skill variation of staff, reduce the pressure of performing manual verification,and improve medical security.
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Objective Non-muscle myosin heavy chain ⅡA (NMHC ⅡA ) plays a significant role in tumor progression and metastasis .Our prior study showed that the expression of NMHC ⅡA was much higher in human bladder cancer sample than that in adjacent tissue .The increased level of NM HC ⅡA expression was correlated with worse prognosis .However ,the role of NMHC ⅡA is unknown in the invasion and metastasis of bladder cancer .Methods RT-PCR and western blotting were used to examine NMHC ⅡA expression lev-els in normal bladder epithelial cells and bladder cancer cell lines .T he migration and invasion ability of cells was tested by wound healing assay and Transwell invasion assay ,respectively .Results Our study showed that knockdown of NMHC ⅡA inhibited migration and invasion in bladder cancer cell line .Conclusion The study indicated that NM HC ⅡA expression increased the invasion and metastasis ability of bladder cancer cell line in vitro .
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Objective To explore the role of LiCl in modulating bacterial-mediated inflammation after Pseudomonas aeruginosa infection.Methods Western-blot was used to determine the efficacy of LiCl usage.The expression of inflammatory cytokines in Pseudomonas aeruginosa-infected macrophages and neutrophils was detected by qPCR.Cell apoptosis was measured by flow cytometry.Results Western-blot data showed that LiCl up-regulated the protein levels of p-GSK-3β(Ser 9)and β-catenin in macrophages and neutrophils,indicating the efficacy of LiCl usage.qPCR data indicated that LiCl enhanced the expression of anti-inflammatory cytokines and suppressed the expression of pro-inflammatory cytokines in Pseudomonas aeruginosa-infected macrophages and neutrophils.Flow cytometry data indicated that LiCl could promoted the apoptosis of Pseudomonas aeruginosa-infected macrophages and neutrophils.Conclusion LiCl inhibited the Pseudomonas aeruginosa-induced inflammation,via regulating the inflammatory cytokine expression and the apoptosis of inflammatory cells.
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Objective:To investigate the clinical application of cognitive behavioral therapy (CBT) and the reasons of the choosing CBT.Method:Totally 14 psychotherapists accepted a semi-structured interview,including their backgrounds of CBT training,the application of CBT in psychotherapy,their attitude toward consultative relations,their opinion on comparing other schools with CBT,and their reasons for choosing CBT.Results:The main reasons for choosing CBT included the influence of important others,the characters of CBT and personal factors.In clinical practice,the most commonly used behavioral techniques included exposure therapy,roll play,relaxation Waining and so on.The most commonly used cognitive techniques included Socratic questioning,cognitive restructuring and challenge unreasonable cognition.Conclusion:Cognitive behavioral therapy (CBT) is suitable for those who are sensible and preferring structural therapy.Nevertheless,the trend in therapy is the integration of different psychotherapy schools.
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Objective:To learn the cognition and attitudes of psychometric ethics of graduate students whose major was clinical psychology or counseling psychology,and to investigate whether ethics training could influence their cognition and attitude.Methods:Researchers distributed the questionnaire online regarding cognition and attitudes of psychometric ethics among clinical and counseling psychology graduate students.Seventy-six feedbacks were collected (4 students were learning ethics courses,27 students had learned ethics courses,45 students hadnt learned ethics courses).The questionnaire included the Psychometric Cognition Questionnaire (PCQ),Situational Judgement Questionnaire (SJQ) and Psychometric Attitude Questionnaire (PAQ).The first two sets of questionnaires reflected ethics knowledge.The last one showed ethics attitudes and behavioral tendencies.Results:The correct rates in participants who had learned or were learning ethics were 76.5％ and 75.1％ at the first and second set of questionnaires,while were 73.9％ and 74.2％ in other participants.The correct rates of three major clauses in PCQ were 90.3％,67.7％ and 74.2％,significantly higher than participants who never learned ethics,whose correct rates were 88.9％,66.7％,and 44.4％ (Ps ＜ 0.05).There were no significant differences in SJQ between those who had learned or was learning ethics and those who had never learned ethics (Ps ＞0.05).Participants who had learned or were leaning ethics scored in sum in PAQ that had no significant differences with those who had never learned ethics(Ps ＞ 0.05).Conclusion:Clinical and counseling psychology graduate students master considerable level of knowledge on ethics and attitude.Ethics training is essential for acquiring psychometric ethics.
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Objective:To investigate the use of psychological testing in hospital and its potential existent ethical issues.Methods:Eleven psychometricians from 6 specialized and 5 general hospitals accepted a semi-structural interview.With (10.8 ± 8.8) years of experiences on average in this field,they tested 2 to 30 patients each day.Content analysis was used to investigate the psychometricians' qualification,usage of tests,informed consent,scoring,explanation and preservation.Results:Normally,the number of psychometricians in one hospital varied from 1 to 4.Out of 11 interviewees,2 were unqualified.Three of the interviewees pointed out there were non-standard circumstances when some doctors choosing the tests.Six of them thought that the frequently used tests' norms were obsolete.Eight interviewees considered that comparatively low psychometric fee influenced their devotion of explaining the results.Conclusion:In hospital,it's necessary to strength the ethical awareness of decision-makers and executors in psychological testing.
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Objective To estimate the methodology for evaluating the analytical measurement range ( AMR) and the clinically reportable range ( CRR) in lab-developed system in clinical chemistry .Methods Method evaluation .Take serum CK for instance , a series of samples were prepared from both a specimen with a high concentration of the analyte of interest and a specimen with a low concentration for the following assays.Average slope method , linear dilution recovery method and the method recommended by Clinical and Laboratory Standards Institute ( CLSI ) EP6-A were used to established AMR in the lab-developed clinical chemistry system.Based on the maximum valid dilution of the specimen and the results of AMR , CRR were determined.One-half of the total error allowance ( TEa) of Chinese health standard was set up as allowance error (7.5%).Results X-Y scatter plot was made by assigning sample numbers to the horizontal axis and actual measured values to the vertical axis , which determined the upper limit of AMR was approximate 1 651 U/L.The results analyzed by average slope method indicated that the linear correlation between expected values and actual measured values was determined , the correlation (r), the intercept (a) and the slope (b) met the linear standard, and AMR was 5-1 699 U/L.The results analyzed by EP6-A indicated that the best fitting curve was obtained by using cubic polynomial method , and the linearity deviation of the minimum concentration was -77.1%, which exceeded one-half of TEa.Followed by the deletion of the maximum concentration , the resumed experiment was done .The results showed that the nonlinear coefficient c of quadratic polynomial and the nonlinear coefficient c and d of cubic polynomial have no significant difference to 0, and AMR was 7.5-1 458.0 U/L.By linear dilution recovery method , the linear correlation between expected values and actual measured values was determined , the correlation ( r) , the intercept ( a) and the slope ( b) met the linear standard , the recovery rates was between 100.0%and 104.8%, and AMR is 5 -1 699 U/L.The CRR was determined to be 5 -33 880 U/L, which met the standard of TEa . Conclusions Average slope method, linear dilution recovery method and EP 6-A method were all used to established AMR in lab-developed clinical chemistry system .Without complicated statistical analysis , linear dilution recovery method was suitable for clinical use .The linearity deviation of the minimum concentration analyzed by EP6-A did not meet the standard of the quality objective system , suggesting defects in the statistical analysis of the results .CRR was feasibly determined by using linear dilution recovery method align with AMR.
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Objective To investigate the relationships between Glucokinase(GCK) gene 4 tag single‐nucleotide polymorphisms , tagSNPs)sites which named rs12702070 ,rs2268569 ,rs2268573 and rs1476891 polymorphisms and type 2 diabetes in Chinese South‐ern Han Population .Methods This study was designed as a case‐control .499 type 2 diabetes patients and 499 healthy controls were chosen .4 GCK tagSNPs sites were analyzed by improved multiple ligase detection reaction(iMLDR) ,and genotype and allele fre‐quency between T2D group and healthy controls could be determined by chi‐square test ,logistic regression analysis ,and tagSNPs were further analyzed under three genetic modes(dominant ,recessive and additive) .What′s more ,Haploview software was used to construct the haplotype of 4 GCK tagSNPs and the linkage disequilibrium(LD) and relationship between various GCK haplotype and T2D susceptibility could be analyzed .Results Genotype distribution of rs2268573 ,rs2268569 and rs1476891 and allele frequen‐cy in T2D group were no significant differences with health controls .Significant differences in genotype distribution of rs12702070 and allele frequency were observed between T2D group and health controls .Under dominant model and additive model ,genotype distribution of rs12702070 in T2D was significantly different from health controls .One LD domain was observed in 3 tagSNPs a‐mong those 4 sites of GCK gene .There are 4 main haplotypes(TAG ,TGG ,TAT ,CGG)in rs2268569 ,rs12702070 and rs1476891 , but all these haplotypes have no relevance to the individual risk of T2D(P>0 .05) .Conclusion The results indicated that the GCK gene tagSNPs site rs12702070 imparts susceptibility to T2D in Han Chinese ,but not rs2268573 ,rs2268569 and rs1476891 .Four main haplotypes in rs2268569 ,rs12702070 and rs1476891 have no relevance to T2D .
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Objective To investigate the precision, trueness, and accuracy of self-developed detection system in clinical chemistry.Methods This was a methodological evaluation.Take serum creatine kinase( CK) for instance, 6 serum sampleswith different leves ( on the upper or lower limit of the reference range or close to the medicine decide levels ( MDLs) , were collected for within-run precision( repeatability) and within-laboratory precision ( intermediate precision ) experiments.5 proficiency testing ( PT ) samples, 5 samples assigned value by reference method, and 40 fresh-frozen serums were measured and compared with reference method for trueness verification.Drawing method evaluation decision chart, calculating total errors and sigma level evaluation experiment based on the CV, bias, and allowed total errors(TEa)were used to evaluate the accuracy performance.The precision, trueness, and accuracy were compared with the quality indicators.Results The within-run precision and within-laboratory precision were less than the highest requirement of Chinese industrial standard.The mean bias was -8.96%, didn′t reachthe required standard (5.5%).After taking corrective actions, all samples but one ( -5.8%) met the required standard. Compared with the reference method, the mean bias on the MDLs was less than TEa.The performance points of the method evaluation decision chart indicated excellence performance.The total errors on MDLs were 14.2%, 10.4% and 7.6%, less than 15%.The sigma levels on MDLs were 5.9, 7.5 and 15, also achieved excellent level. Conclusions The precision, trueness, and accuracy performance of CK measured by self-developed detection system achieved excellent level of the Chinese industry standard, and the same results were found from different evaluation methods.
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Objective To establish TaqMan probe real‐time Fluorescent Quantitative PCR in detecting Cryptococcus neoformans genomic DNA ,and to provide important method for detection of cryptococcal meningitis .Methods According to the Cryptococcus neoformans ITS‐rDNA sequences obtained from NCBI ,specific primers and probe were designed based on the conserved sequences , a specific 114 bp fragment was amplified by primers and probe ,then recombinant plasmid was constructed .Eight different concen‐trations from 1 .42 × 10 copy/μL to 1 .42 × 10 copy/μL were used as templates by 2 μL .In the optimum reaction condition ,the sen‐sitivity ,specificity and repeatability were evaluated and standard curve was established .15 clinical cryptococcal meningitis strains i‐solated from clinical diagnosis patients were detected .Results The real‐time PCR showed high sensitivity and specificity and was a‐ble to detect 2 .84 × 102 copies of plasmid DNA .The detection sensitivity was 2 .84 × 102 copies plasmid DNA by real‐time PCR ,no amplification curve was detected with human genomic DNA ,other fungus ,bacterias and viruses .The CV of inter‐assay were 2 .86% ,1 .48% and 1 .36% respectively with excellent reproducibility .Fifteen clinical isolated strains could be detected accurately . Conclusion A method of detection of Cryptococcus neoformans DNA by real‐time PCR is established successfully with high sensi‐tivity and specificity ,and the results are stable ,could diagnose cryptococcal meningitis rapidly and early .
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Objective To investigate the relationships between glucokinase(GCK) gene 3 tag single‐nucleotide polymorphisms (tagSNPs)sites rs2971672 ,rs2268573 and rs2300587 polymorphisms with type 2 diabetes (T2DM ) .Methods A total of 499 south‐ern Han inpatients with T2DM (T2DM group) in our hospital and contemporaneous 499 Han individuals undergoing the physical examination(control group) in the Health and Fitness Protection Center of our hospital from August 2013 to December 2014 were chosen .The GCK gene 3 tagSNPs sites in all subjects were genotyped by adopting the improved multiple ligase detection reaction (iMLDR) ,and the genotype and allele frequency between the T2DM group and healthy controls were compared by the chi‐square test ,logistic regression analysis ,moreover the tagSNPs sites were performed the correlation analysis under three genetic modes (dominant ,recessive and additive) .The Haploview software was used to construct the haplotype of GCK gene 3 tagSNPs and the linkage disequilibrium(LD) and relationship between various GCK haplotype and T2DM susceptibility was analyzed .Results The differences of rs2268573 and rs2300587 genotypes(χ2 =3 .361 ,2 .076 ,P>0 .05) and allele frequency(χ2 =0 .222 ,1 .980 ,P>0 .05) between the T2DM group and the control group were not statistically significant .The difference of rs2971672 genotype(χ2 =6 .896 , P<0 .01) and allele distribution(χ2 =4 .708 ,P<0 .05) between the T2DM group and the control group was statistically signifi‐cant .Under the dominant genetic model and additive genetic model ,the genotype distribution of rs2971672 between the T2DM group and the control group was statistically significant(OR= 1 .74 ,95% CI:1 .17 -2 .57 ,P<0 .01 ;OR=1 .51 ,95% CI:1 .06-2 .14 ,P<0 .05) .Among 3 GCK gene sites ,rs2971672 and rs2300587 had the LD domain including 3 main haplotypes of TC ,TA and CA3 ,the TA and CA haplotypes all decreased the risk suffering from T2DM(OR=0 .81 ,95% CI:0 .66-1 .00 ,P<0 .05 ;OR=0 .78 ,95% CI:0 .62-0 .98 ,P<0 .05) .Conclusion In Han population ,GCK gene rs2971672 site is closely related with T2DM ge‐netic susceptibility ,while rs2268573 and rs2300587 sites have no obvious correlation with T2DM susceptibility .Haplotype TA and CA in rs2971672 and rs2300587 LD domain all reduce the individual risk suffering from T2DM .
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Objective To explore the role of LiCl in microbial suppurative keratitis .Methods Western-blot assay was used to detect the efficacy of LiCl .Inflammatory cytokine levels were examined using Real-time PCR .Cell apoptosis was detected by using TUNEL and flow cytometry assays .Results LiCl promoted corneal resistance to PA infection .Real-time PCR data showed that LiCl enhanced IL-10 expression ,but suppressed TNF-expression .TUNEL and flow cytometry data showed that LiCl promoted the apoptosis of infiltrating cells .Conclusion LiCl promoted host resistance against microbial suppurative keratitis ,via regulating in-flammatory cytokine expression and cell apoptosis .
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Objective To explore the relationship between glucose-6-phosphate dehydrogenase deficiency and type 2 diabetes and to provide reference for clinical diagnosis,monitoring and treatment of Type 2 diabetes.Methods Subjects were selected from our hospital,including 173 cases of healthy volunteers assigned to the control group;1 95 cases with type 2 diabetes conforming to the diagnostic criteria of WHO were assigned to the diabetic group.97 cases were diagnosed with glucose-6-phosphate dehydrogenase deficiency in our hospital,of whom 82 cases were assigned to simple glucose-6-phosphate dehydrogenase deficiency group,and 1 7 cases were assigned to the diabetes with glucose-6-phosphate dehydrogenase deficiency group.The correlation of glucose-6-phos-phate dehydrogenase activity and diabetes was measured by each detected value.Results Compared with the control group,the glu-cose-6-phosphate dehydrogenase activity and RBC count in diabetic group were higher(P <0.05).Positive correlations between glu-cose-6-phosphate dehydrogenase activity and RBC count in the two groups were significant(P <0.05).HbA1c and FBG showed a significant positive correlation in the control group,diabetic group and diabeties with glucose-6-phosphate dehydrogenase deficiency group(P <0.05).But there was no significant correlation in the glucose-6-phosphate dehydrogenase deficiency group.The rate of screening for glucose-6-phosphate dehydrogenase deficiency in diabetes group was 3.6%,and the rate in the control group was 1. 1%.When HbAlc≥6.5%,the rate of screening for glucose-6-phosphate dehydrogenase deficiency in the diabetes group was signifi-cantly higher than that in the control group (χ2 =4.239,P =0.039).Conclusion The level of HbA1c is discordant with that of blood glucose in diabetic patients with glucose-6-phosphate dehydrogenase deficiency group.Diabetes leads to the increasement of the rate of glucose-6-phosphate dehydrogenase deficiency.The glucose-6-phosphate dehydrogenase activity of diabetic patients with non-glucose-6-phosphate dehydrogenase dificiency is higher than that of the normal group.It may be associated with the level of RBC or increase of compensatory.Further more,glucose-6-phosphate dehydrogenase activity may be a good indicator for controlling diabetes,which remains to be further studied.
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Objective To estimate the prevalence of Glucose‐6‐phosphate dehydrogenase(G6PD) deficiency in Zhongshan area . Methods The activity of G6PD in red blood cells was determined by using ultra‐violet rate method for neonates ,couples of child‐bearing age and suspected patients who had clinical symptoms in Zhongshan area from 2012 to 2013 .Results The total detection rate of G6PD deficiency was 4 .37% (1 030/23 595);in male the detection rate was 9 .42% (513/5 447);in female the detection rate was 2 .85% (517/18 148) .Conclusion The incidence of G6PD deficiency were high in Zhongshan area .Therefore ,more attention should be paid to the screening of the disease in neonates and couples of childbearing age so as to reduce the incidence of G6PD defi‐ciency and prevent the complications caused by the disease .
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Aim To analyze differential expression and interaction of β-catenin/ICAT proteins in HL60 cells when they were induced into monocytic differentiation, and to figure out the mechanism of NSC67657 in cellu-lar induction. Methods HL60 cells were treated by 10 μmol · L-1 NSC67657 , and cellular differentiation could be observed by cytochemical staining and flow cytometry. Then, RT-PCR and Western blot were em-ployed to determine the differential expression of β-catenin/ICAT genes and proteins. Co-immunoprecipi-tation assay was used to confirm the interaction of β-catenin/ICAT proteins, and laser co-focus light mi-croscopy technology was used to co-indentify proteins differential expression and intracellular location. Re-sults HL60 cells could be induced into monocytic dif-ferentiation after 5 days treatment using 10μM NSC67657 . The CD14 ( +)% cells could be up to o-ver 90%, and cytochemical staining reports were con-sistent with this result. The expressions of ICAT gene and protein were up-regulated significantly ( P <0. 01 ) , but the expressions ofβ-catenin gene and pro-tein, on the contrary, were down-regulated(P<0. 05) when HL60 cells were induced into monocytic differen-tiation. From co-immunoprecipitation assay findings, ICAT protein interacted with β-catenin protein, and the absorbance of protein electrophoresis bands in-creased in differentiated cells. From laser co-focus light microscopy assay findings, the fluorescence of ICAT and β-catenin protein could be both observed in cytoplasm and nucleus. In drug treated HL60 cells, the fluorescence of ICAT protein was enhanced both in cytoplasm and nucleus, however, the fluorescence ofβ-catenin protein, which looked like transferring into different organelles, decreased significantly in nucleus, but increased in cytoplasm. Conclusions HL60 cells could be induced into monocytic differentiation by NSC67657 and β-catenin/ICAT proteins differentially expressed during cellular differentiation. The enhanced interaction of β-catenin/ICAT proteins and β-catenin protein transferring from nucleus into cytoplasm indi-cates that NSC67657 probably induces HL60 cells into monocytic differentiation through down-regulating β-catenin protein and blockingβ-catenin protein from nu-cleus.
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Objective To explore the feasibility of measurement uncertainty in complete blood count using internal quality con-trol(IQC) data associated with Sysmex Network Communication System(SNCS) comparative data .Methods Complete blood count assay including white blood cell(WBC) count ,red blood cell(RBC) count ,hemoglobin(Hb) determination ,hematocrit(HCT) values and platelet(PLT) count were involved to evaluate measurement uncertainty according to the instruction of CNAS-TRL-001 .Meas-urement uncertainty evaluation was established by internal measurement reproducibility using IQC data ,and standard measurement uncertainty by bias using proficiency testing(PT) data and SNCS data ,followed by relative combined standard measurement uncer-tainty and relative expanded measurement uncertainty was calculated .Meanwhile ,the measurement uncertainty was compared by u-sing PT data and SNAS comparative data .Results Relative expanded measurement uncertainty of the above mentioned index by u-sing IQC data associated with PT data was the following :WBC(10 .02% ,7 .24% ,7 .04% ,from level 1 to level 3 respectively) ,RBC (2 .40% ,1 .72% ,1 .92% ,from level 1 to level 3 respectively) ,Hb(3 .54% ,2 .56% ,2 .50% ,from level 1 to level 3 respectively) , HCT(4 .12% ,3 .18% ,2 .86% ,from level 1 to level 3 respectively) ,PLT(15 .36% ,8 .86% ,7 .94% ,from level 1 to level 3 respec-tively) .Relative expanded measurement uncertainty of the above mentioned index by using IQC data associated with SNCS compar-ative data was the following :WBC(11 .66% ,7 .34% ,6 .40% ,from level 1 to level 3 respectively) ,RBC(2 .26% ,1 .60% ,1 .64%from level 1 to level 3 respectively) ,Hb(3 .36% ,2 .36% ,2 .10% ,from level 1 to level 3 respectively) ,HCT (3 .36% ,3 .04% , 3 .18% ,from level 1 to level 3 respectively) ,PLT (13 .34% ,8 .36% ,7 .14% ,from level 1 to level 3 respectively) .Conclusion Measurement uncertainty in complete blood cell could be estimated by using IQC data associated with SNCS comparative data , which is in accord with the instrument of target measurement uncertainty .