Objective To analyze the relationship between anemia and blood levels of lead and trace elements in children,and to provide basis for the prevention and treatment of anemia.Methods 80 children with anemia(case group)and 90 health children (control group)were enrolled in the study.The blood levels of trace elements (iron,zinc,calcium,magnesium,copper)and lead were detected in all subjects,in order to analyze their correlations with anemia.Results Compared with control group,the blood levels of iron,zinc and magnesium in case group were significantly lower(P<0.05),while the blood levels of calcium and lead were significantly higher(P<0.05).The blood level of copper in case group was close to that in control group(P>0.05).With the re-duction of iron,zinc and magnesium and the rise of calcium and lead,the severity of anemia increased significantly(P<0.05). Effects of copper level on the severity of anemia was not obvious(P>0.05).The blood level of iron was positive correlated with Hb,RBC and MCH (P<0.05).The blood level of zinc was positive correlated with Hb and MCH(P<0.05).The blood level of lead was negative correlated with Hb,MCV and MCH(P<0.05).Conclusion The blood level changes of trace elements and leas effect the severity of anemia in children.It is necessary to test the blood levels of trace elements and lead.

Objective To evaluate the performance of enzyme linked immunosorbent assay(ELISA)reagent kit of hepatitis B surface antigen(HBsAg)with Clinical and Laboratory Standards Institute(CLSI)document EP12-A2,and to compare two different manufacturer kits′performance.Methods According to CLSI document EP12-A2,to evaluate the performance of two different manufacturers′HBsAg ELISA kits.Results The positive rates both were ≥95% by using the kits of two manufacturers to detect the same sample with 120%of the critical concentration.The negative rates were both ≥95%by using the kits of two manufactur-ers to detect the same sample with 80% of the critical concentration.The intra assay coefficients of variation were ≤15%,and the interassay coefficients of variation were ≤20%.The sensitivities of two manufacturers′kits were respectively 94.2%(95%CI:84.3%-98.0%)and 92.3%(95%CI:81.8%-97.0%),and the specificities were 98.0%(95%CI:89.5%-99.6%)and 100.0%(95%CI:92.8%-100.0%).The 95%interval of sensitivity of two manufacturers′s kits was 5.46%-10.19%,and which of specificity of two manufacturers′s kits was -2.00%-5.32%.Conclusion The kits of two manufacturers can provide stable re-sults of HBsAg detection with concentrations ±20% outside the critical concentrations at the cutoff point with high sensitivity and specificity.Both of the kits have no significant differences in sensitivity and specificity.

Objective To analyze and approach the optimizing of ELISA assay for HBV serum markers.Methods To compare the critical value rate of flushing with distilled water,flushing with cleaning tap water and flushing without anything.Results The HBsAg critical value rates of flushing with distilled water,flushing with cleaning tap water and flushing without anything were 0.52%,2.60% and 5.56%.The HBsAb critical value rates of flushing with distilled water,flushing with cleaning tap water and flushing without anything were 5.52%,8.02% and 14.86%.The HBeAg critical value rate of flushing with distilled water,flush-ing with cleaning tap water and flushing without anything were 2.82%,2.92%and 3.55%.Conclusion Flushing with distilled wa-ter or flushing with cleaning tap water can reduce the high critical value rate because of enzyme or developer pollution.Flushing with distilled water exhibits more efficacy than flushing with cleaning tap water.

Objective To evaluate the clinical applicability of Gold Lebel,ELISA and LiCA,through detecting HBsAg by all the three methods.Methods A total of 1 000 sapmles were selected,and each sample was detected HBsAg by using three methods. Each positive sampale was detected repeatedly to identify false negative and false positive.Results In all the three methods,the most sensitive and specific method was LiCA,next was ELISA,and the last was Gold Lebel.But Gold Lebel method was more sim-ple and rapid for preliminary screening for emergency or preoperative patients with its sensibility of 1 ng/mL.ELISA had too many operating steps to bring false negative and false positive,while its high sensitivity and specificity and low price might be accepted by commonage and fit for mass simples.LiCA′s especially high stability,accuracy,easily operating could better reflex HBV replication and therapy effects of antiviral drugs in spite of its slight cost.Conclusion The suitable HBsAg detection method should be chosen according to clinical demands.

Objective To investigate the RHD gene polymorphism among the RhD negative Han individuals in Xuzhou.Methods The RhD antigen phenotypes were detected by routine serological techniques,and were identified with indirect antiglobulin test (IAT).The genotypes of RHD were detected by using sequence specific primer polymerase chain reaction(SSP-PCR).Results A-mong 110 RhD negative individuals,there were 5 cases carring RHD positive gene,47 cases carring RHD negative gene,22 cases carring RHD-CE(2-9)-D gene,17 cases carring DVI Ⅲ gene,2 cases carring weak D15 gene,and 17 carring DEL-1227A gene,re-spectively.Conclusion The RhD negative Han individuals present complex RHD gene polymorphism in Xuzhou region,and variant alleles such as RHD-CE(2-9)-D,DVI Ⅲ,DEL-1227A are given priority.

Objective To evaluate the diagnostic value of procalcitonin (PCT)in bacteremia.Methods Clinical data of adult pa-tients with suspected bacteremia were retrospectively analyzed for white blood cell counts(WBC),neutrophil absolute (NEU)and PCT levels in blood samples collected at the first time of blood culture.Receiver operating characteristic (ROC)curve was used to evaluate the values of different indexes for the prediction of bacteremia.Results A total of 120 patients were enrolled,including 47 patients (39.2%)with bacteremia and 73 patients (60.8%)without bacteremia.PCT,WBC and NEU levels were significantly higher in bacteremia patients than patients without bacteremia (P<0.01).Area under ROC curve of PCT was 0.836 (95%CI:0.767-0.904),which was significantly higher than WBC (0.676,95%CI:0.676-0.600)and NEU (0.696,95% CI:0.696-0.617).While area under ROC curve of WBC and NEU were no significantly different (P>0.05).Taking 0.43 ng/mL as cutoff value,the negative predictive value of PCT was 94.7%.The best cutoff value of PCT was 2.27 ng/mL,which was associated with sensitivity of 74.47%,specificity of 90.00%,positive predictive value of 83.3% and negative predictive value of 84.0%.Conclusion PCT might be with more clinical value for the prediction of bacteremi than WBC and NEU.

Objective To investigate the association between the length of time-to-ejaculation and the quality of semen obtained by masturbation.Methods Ejaculates from 284 men under infertility assessment were analyzed according to World Health Organi-zation guidelines.Seminal neutralα-glucosidase (NAG)、zinc(Zn)and fructose(FRU)were also measured.All patients were divid-ed into three groups according to the length of time-to-ejaculation,including group A with time-to-ejaculation not more than 10 min, group B with time-to-ejaculation of 11-15 min and group C with time-to-ejaculation more than 15 min.Statistical analysis was per-formed.Results Time-to-ejaculation was negatively correlated with sperm concentration (r= -0.20,P= 0.02),total sperm count (r= -0.20,P= 0.04),NAG (r= -0.20,P= 0.01),total count of progressive motility (r= -0.40,P= 0.03)and fructose (r=-0.30,P= 0.02),respectively,but not correlated with age,sexual abstinence,semen volume,sperm motility and Zinc.Sperm con-centration,total sperm count and total count of progressive motility in were higher than group B and C (P<0.05),and fructose was higher than group C (P<0.05).Conclusion Time-to-ejaculation might be associated with semen parameters.Acute stress during semen collection via masturbation at a clinic could infect the quality of semen.

Objective To construct a simple method for the measurement of activity of S-homocysteine methyltransferase (HMT),and explore the best processing condition for HMT and the preservation of HMT.Methods HMT was expressed in pro-karyotic system by using genetic engineering technology,then was purified by using affinity and Sephadex G1 5 chromatography. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE)was performed to identify the physicochemical and bio-logical properties of target protein.Based on the principle of 5 ,5′-disulfide-double (2- nitro benzoic acid)(DTNB)could react with sulfydryl compounds rapidly,the reduction of homocysteine was detect to evaluate the activity of enzyme,then the best processing conditions of HTM were determined.Activity of enzyme,preserved in preservation solution with or without glycerol and preserved under different temperatures,was detected.Activity remaining ratios were detected and compared between HMT preserved in pres-ervation solution with different protective agents of different concentration and preserved by cryodesiccation.Results The purity of recombined HMT was above 95%,with molecular weight of 36 000 and excellent catalytic activity,and the catalytic activity was 2 000 U/mg.The optimum condition for the detection of biological activity was using HEPES buffer of pH 7.4 at 37 ℃ and reac-tion for 25 min.Glycerol could significantly prolong the preserving time of HMT,and half activity of HMT could be remained for six months.The reservation rates of activity of HMT,preserved in preservation solution with mannitol and trehalose,were 104%and 100%,respectively.Conclusion HMT could be obtained through genetic engineering.A simple test method of HMT was es-tablished,and the best processing conditions and preserving methods of HMT were determined,which laid a foundation for clinical application.

Objective To express and preliminary identify scavenger receptor B2 (SCARB2)protein.Methods SCARB2 cDNA was amplified by reverse transcription-polymerase chain reaction (RT-PCR)from mRNA extracted from RD cells,and cloned into pMD19-T vector.Expression vector pET28a(+)/SCARB2 was constructed,and SCARB2 protein was expressed in prokaryotic ex-pression system.Obtained recombinant proteins were identified by detection of Western blot using His labeled monoclonal antibody.Results Recombinant SCARB2 proteins,expressed by induction of isopropy-β-D-thiogalactoside (IPTG),were success-fully purified and specifically recognized by His labeled monoclonal antibody.Conclusion SCARB2 proteins could be expressed and identified successfully,which could provide reference for researching mechanism of enterovirus type 71 (EV71)and scavenger re-ceptor protein,and for preparation of SCARB2 monoclonal antibodies.

Objective To investigate the clinical value of homocysteine (Hcy)and high-sensitivity C-reactive protein (hs-CRP) in diagnosis and treatment of senile angina pectoris.Methods According to clinical curative effect of atorvastatin treatment,senile patients with angina pectoris were divided into high efficient group,effective group and ineffective group.Level of Hcy and hs-CRP,detected before and after treatment,were compared,and their correlation with creative effect were also analyzed.re and after treatment in the group.Results Levels of Hcy and hs-CRP were correlated with the curative effect of senile patients with angina pectoris,and the sensitivity and specificity of Hcy were higher than hs-CRP.Conclusion Hcy and hs-CRP could be used to as sen-sitive indicator for the diagnosis of senile angina pectoris,which might be also very important for the evaluation of curative effects.