Objective To analyze and evaluate the performance of the double antigen sandwich method reagents by comparing the detection results of three kinds of domestic hepatitis C virus(HCV)antibody detection kits with indirect method and sandwich method.Methods The different test methods were designed.The HCV antibody detection by using three kinds of reagents was performed on the collected positive and negative specimens and those specimens with positive reaction by the single and dual rea-gents were simultaneously performed the recombinant immunoblot assay(RIBA)and the nucleic acid detection;in addition,after the multiple proportion dilution,the quality control material with known content was parallelly detected with three kinds of reagents. All the detection results were conducted the comparison and the comprehensive analysis.Results Compared with the confirmation reagent,the false positive rates of two kinds of indirect method reagent and one kind of sandwich method reagent were 40.9%, 59.1% and 2.2% respectively.Their positive rates were 100%,but the analytical sensitivity of the sandwich method was higher than that of the indirect method by 1 ~4 times.Conclusion The sandwich ELISA reagent is significantly superior to the indirect EllSA reagent in the aspects of sensitivity and specificity.

Objective Human A20 is one of the endogenetic regulating proteins of inflammation in vivo and play important roles in preventing uncontrolled inflammatory response as well as endogenetic cellular protection.Our aim is to prepare a recombinant protein of human A20(rhA20) in Pichia pastoris and then research its roles in sepsis therapy.Methods rhA20 yeast expression vector yevCFP-hA20 was constructed by genetic engineering.Pichia pastoris with linearized yevCFP-hA20 was electroporated.The high copy clones were screened with MD/His~(-) medium and YPD/G418 gradient medium.The number of hA20 in yeast genome was detected by quantitative PCR.The rhA20 was expressed by methanol induction and identified by SDS-PAGE and Western blotting.The optimum conditions(including pH and ingredient of induced medium as well as the temperature of induced culture) were investigated.Results The molecular weight of rhA20 is about 93?10~3.After expression condition's optimization by decreasing the pH of BMMY medium,adding a low dosage of EDTA and the substrate of protease,as well as decreasing the induced temperature,the full expressing products was up to 11 times,rose to 18.24% in total protein and achieved to 254.1 024.52?g/ml.Conclusion we have successfully expressed rhA20 in Pichia pastoris GS115.The expression amount of full rhA20 increases clearly by progressing the ingredient and pH of BMMY and using a lower inducing temperature.This starts the biosynthesis of hA20 in vitro.A new solving regime is provided to other biologist when they meet product's degradation by using Pichia pastoris GS115.

Objective Effects of methylprednisolone on observation of intramedullary spinal cord tumor treatment.Methods Through the 9 cases of intramedullary spinal cord tumors in patients after adjuvant therapy of methylprednisolone clinical observation and analysis.Results 9 cases of ostoperative patients after treatment with methylprednisolone,his condition to resume soon,but the response to 5 cases of side effects,as symptomatic treatment can be back to normal.Conclusions Application of adjuvant therapy of methylprednisolone after spinal marrow tumor effects although good,it should be noted that the occurrence of side effects,and do:the right of dispensing,close observation and timely manner.

The external and internal characteristics of domestic master and doctorial papers on rheumatoid arthritis were analyzed by visualization analysis, which revealed the publication of and hotspots and frontiers in domestic master and doctorial papers on rheumatoid arthritis, and can thus provide reference for the development of medical information resources and recent advances in rheumatoid arthritis for the broad masses of scientific workers.

Objective To study the change of the peripheral blood cells of soldiers in high altitude hypoxia,to analyze its influ-ence on the blood cells.Methods The blood cell were compared among the soldiers in the plains,entered plateau after 6 months and stationed plateau for a long time,and to analyzed the effect of high altitude hypoxia environment on blood cells.Results WBC, RBC,hemoglobin,hematocrit and mean corpuscular hemoglobin concentration were obviously above plain group and long staff group (P <0.05),and mean corpuscular volume was obviously lower than those of plain group and long staff group (P <0.05).White cell and lymphocytes in plateau group were also obviously higher than those in plain group (P <0.05),and long staff group no signifi-cantly differences (P >0.05).platelet,mean platelet volume,platelet distribution width was clearly higher than that of plain group (P <0.05 ).Conclusion Blood cells of soldiers training in the high altitude hypoxia environment are significantly increased,and changes in blood cells are closely related to emplacement time and altitude.

Objective To analyze results of blood test for first‐time blood donation volunteers and mutual‐aid blood donors in Anyang area ,in order to improve cognition of characteristics of different blood donors and safety of blood transfusion .Methods A total of 31 363 cases of first‐time blood donation volunteers(first‐time blood donation group) and 1 046 cases of mutual‐aid blood donors(control group) from January to December 2013 in Anyang area were collected .The results of blood tests were compared and related factors affected qualities of blood were analysed .Results A total of 1 160 cases of donors in the first‐time blood donation group were failed to pass the blood test and the unqualified rate was 3 .70% ,in the control group 17 cases of donors failed to pass the blood test and the unqualified rate was 1 .62% ;the difference was statistically significant (P<0 .05) .The test results of unqual‐ified blood samples shown that in the first‐time blood donation group the constituent ratios of alanine aminotransferase(ALT) ,anti‐body to hepatitis C virus(anti‐HCV) ,antibody to human immunodeficiency virus(anti‐HIV) ,hepatitis B surface antigen(HBsAg) and syphilis positive blood donors were 37 .67% ,12 .59% ,8 .79% ,31 .38% and 9 .57% respectively ,and those in the control group were 29 .41% ,17 .65% ,5 .88% ,35 .30% and 11 .76% respectively .The constituent ratio of ALT positive blood donors between the two groups had statistically significant difference(P<0 .05) .There were statistically significant differences in gender ,age and occu‐pations between unqualified blood donors in the two groups(P<0 .05) .Conclusion The first‐time blood donation volunteers might have relatively higher unqualified rate in blood test ,it is necessary to enhance blood screening and management before transfusion , in order to ensure safety of blood transfusion .

Objective To investigate relationship of single nucleotide polymorphism (SNP)at rs5029924 locus in A20 promoter region and posttraumatic sepsis.Methods PCR-DNA sequencing was used to analyze different gene distribution at rs5029924 locus of 103 trauma patients with sepsis (Group A),120 trauma patients without sepsis (Group B) and 135 healthy peoples (control group).Relation of different genotypes at rs5029924 locus to sepsis susceptibility was analyzed.Peripheral blood cells of healthy peoples of different genotypes were stimulated using lipopolysaccharides (LPS) in vitro.Expression of A20 mRNA was measured by fluorescent quantitative PCR,expression of A20 protein by flow cytometry,and levels of TNF-α and IL-1β by ELISA method.Results Frequency of rs5029924 genotypes CC,CT and TT was respective 77.8％,20.0％ and 2.2％ in control group； 63.1％,34.0％ and 2.9％ in Group A； 83.3％,15.0％ and 1.7％ in Group B.Significantly lower frequency of CC genotype was observed in Group A when compared to Group B and control group (P ＜0.05),but no statistical differences were recorded between Group B and control group (P ＞ 0.05).CT/TT genotype increased risk coefficient of sepsis to 2.397-fold higher level when compared to CC genotype.Allele T increased prevalence of sepsis significantly as well (OR =2.056) when compared to allele C.After LPS treatment in vitro,CC genotype individuals revealed significantly higher levels of A20 mRNA and protein in peripheral blood leukocytes,but significantly lower levels of TNF-α and IL-1 β when compared to CT/TT genotype individuals.Conclusion Polymorphism of rs5029924 locus is associated with sepsis susceptibility and the reason may be that mutant genes affect promoter activity and down-regulate A20 expression,which fails to suppress inflammation.

Flow cytometric immunophenotyping has evolved from two-parameter quantitative measurement of peripheral blood lymphocytes to five-or more parameter qualitative evaluation of bone marrow and lymph node in hematopathology.Leukemia/lymphoma immunophenotyping represents an important addition to histomorphology in the diagnosis,classification and monitoring of hematolymphoid neoplasms.The complexity of five-or more parameter analyses and the interpretation of the data rely on standardization and validation of the instrument,the reagent and the procedure.In addition,clinical flow cytometry laboratories in U.S.A are required to document proficiency testing,sample preparation as well as accuracy,specificity,sensitivity and precision of methodology.CLSI and UCCC recommend that each laboratory should validate its own qualitative and quantitative procedures.This paper introduces the procedures for validation and quality control in a clinical flow cytometry laboratory in the United States.

Courseware design of Flow cytometry and its clinical application includes aim,contents,methods,teaching effect,key problem and solving measures,etc.It aims to standardize teaching content and overcome the contradiction between more teaching content and less teaching time.It is also an effective way to save limited teaching expense and contributes to making the course of Flow cytometry more concise,lively and concrete.So it can increase overall teaching quality and level.

Objective To investigate the relationship between the single nucleotide polymorphisms (SNP) of rs1008438 in HSP70 promoter and the susceptibility of high altitude pulmonary edema (HAPE) .Methods The PCR‐DNA sequencing method was used to analyze gene distribution of rs1008438 in 100 HAPE patients and 200 healthy people ,and the relationship between dif‐ferent genotypes and HAPE was evaluated .Meanwhile ,HSP70 protein in cytoplasm and nuclei of white blood cells were detected by ELISA in patientgroup and healthy group .TNF‐α,IL‐1β and IL‐6 levels were analyzed by protein chip technology and EVI‐DENCE180 automatic chip reader .Results The TT ,GT ,and GG genetype frequencies of rs1008438 in HAPE patients and healthy controls were 76 .0% ,20 .0% ,4 .0% ,and 93 .0% ,6 .5% ,0 .5% ,respectively .The TT genetype frequency in HAPE patients was significantly lower than that in healthy control(P<0 .05) .The HAPE incidence rate in GT/GG genetype was 4 .195 times higher than that in TT genetype ,and the allele G can also significantly increase the prevalence of HAPE compared to T allele (OR=4 .178) .ELISA showed that HSP70 were higher in HAPE group of all genotypes than those in healthy control of the same geno‐type .And the nuclei/cytoplasm ratio of HSP70 in TT genotype was higher than that in GT/GG genotype .Protein chip showed that the levels of TNF α,IL‐1β,and IL‐6 in HAPE group were significantly higher than those in healthy control .Conclusion The poly‐morphism of rs1008438 is related to susceptibility of HAPE .Mutate genetype may change the promoter activity and increase the ex‐pression of HSP70 ,which induced HAPE .