Abstract:Membrane-associated guanylate kinase family proteins mediate the clustering and aggregation of ion channels,receptors and cell adhesion molecules via their PDZ、SH₃、GK domains,respectively. MAGUKs construct the cytoskeleton, take part in the signal transduction and modulate the cell cycle process,as well as the neural development. The characters of MAGUKs'structure ,their functions and some members of the family were explicated.

Abstract:Inhibition at central synapses is majorly mediated by glycine and γ-aminobutyric acid (GABA). Glycine acts by binding to a specific receptor and opening an intrinsic chloride channel. The inhibitory glycine receptor (GlyR) widely expressed in many regions of the central nervous system is a member of the ligand-gated ion channel receptor superfamily. GlyR consists of five similar subunits (3α, 2β) arranged to form a ring around a central pore. Since the glycine-binding site is distant from the pore, long-range allosteric interactions are needed to couple agonist binding to channel gating. Recent advances in understanding the structure, physiological and pharmacological characteristics of GlyR are reviewed and modulation of GlyR and possible underlying mechanisms are discussed, in special reference to the recent work in our laboratory.

Abstract:Since Prusiner, suggested the prion hypothesis in 1982, a wealth of experiments have supported it to be true. However, the function of the cellular prion protein(PrP^C) remains unclear. But recently evidence is showing that PrP^C could specifically bind to Cu²⁺ and may transport Cu²⁺ to SOD1 by endocytosis from the plasma membrane via clathrin-coated pits so that taking part in copper metablism. Moreover, other evidence also shows that Cu²⁺ could enhance the reversibility of denatured PrP^Sc and may determine the difference of some PrP^Sc strains.

Abstract:The class 1 release factor is required in termination of protein synthesis. It could accurately recognize the stop signal and promotes the hydrolysis of the ester bond linking the polypeptide chain with the peptidyl (p)site tRNA. The model of “molecular mimicry between release factor and tRNA” explains the resemblance of function. Its highly conserved, universal GGQ motif and a tripeptide ‘anticodon’ play important roles in translation termination respectively.

Abstract:Vascular endothelial growth factor (VEGF) plays a central role in tumor angiogenesis. It stimulates endothelial cell proliferation and vessel hyperpermeability, promotes cell migration, and inhibits apoptosis. All these actions of VEGF are mediated by receptor tyrosine kinase, vascular endothelial growth factor receptor (VEGFR). Selective targeting VEGFR signal transduction pathway may be proved to be useful in developing tumor angiogensis inhibitors.

Abstract:Uteroglobin (UG) is a steroid-inducible, evolutionarily conserved, multifunctional protein. In terms of molecular structure, regulation and synthesis mechanism, UG, which mediated autocrine and paracrine via its putative receptor, was surmised to be a member of the novel cytokine/chemokine family. Clarification of the structure, regulation and function of UG would contribute to elucidating human common disease, exploring the mystery of embryo implantation and researching the mechanism of oncogenesis.

Abstract:Chicken anemia virus vp3 gene encodes a small protein—Apoptin. It can induce apoptosis in tumorgenic or transformed cells but not in normal cells through a way that is independent of p53 reaction and can not be suppressed by the excessive expression of Bcl-2. All these facts make apoptin a potential antitumor agent. The unique way of apoptin inducing apoptosis is also useful to the study of the mechanisms of cell transformation and pathways of apoptosis.

Abstract:The concentration of calcium ion in cytoplasm is in strict control. Disequilibrium of calcium homeostasis will result in severe injury even death of cells. The roles of such disequilibrium in cells death resulted from exogenous factors, evidences of direct lethal effect of the disequilibrium on cell and effects of calcium ion in apoptosis were reviewedd. Mechanisms of these effects were discussed. In the end, on the basis of summary, a new way to inhibit cancer cell was suggested, by inducing disorder of calcium homeostasis selectively in cancer cells.

Abstract:DNA ligases are important enzymes of DNA metabolism. The reaction of nicked DNA joining catcalyzed by the DNA ligase is required in DNA replication and in DNA repair pathways that require the re-synthesis of DNA.The recent advances in isozyme DNA ligase are summarized: bacteria DNA ligases are all NAD^＋-dependent and their sequences show a considerale degree of similarity and all are approximately the same size (～75 ku). All known eukaryotic DNA ligases are powered by ATP. DNA ligase Ⅰ is required for Okazaki frament joining and some repair pathways; DNA ligase Ⅱ appears to be a degradation product of ligase Ⅲ; DNA ligase Ⅲ has several isoforms, which are involved in repair and recombination of DNA.

Abstract:About 35 selenoproteins have been identified and characterized, though many have roles that have not yet been fully elucidated. Selenocysteine represents the 21^st amino acid which is encoded by the UGA triplet in selenoproteins mRNA. Incorporation of selenocysteine in selenoproteins is rather complex but has been widely elucidated in prokaryotes. Four gene products (SELA, SELB, SELC, and SELD) and a specific stem-loop secondary structure which is termed selenocysteine insertion sequence (SECIS element) are required. However, the biosynthetic pathway of selenoproteins in eukaryotes may proceed by different routes in such aspects as the position and structure of SECIS element, specific elongation factors and other RNA-RNA or RNA-binding protein factors interactions. Observations also showed that translation of UGA as Sec in mammalian cells was an inefficient process and the regulation of the same UGA codon existing in an identical position in mRNA serves different functions (stop codon and Sec codon) might be involve in this process.

Abstract:Large amount of genome data has been obtained from the rapid progress of genome sequencing. Along with this trend, many new approaches for the study of protein function have been invented. A brief introduction and discussion of those methods such as domain fusion analysis, protein phylogenetic profiles, cluster analysis, protein structure analysis, insertional mutagenesis and the combined algorithm for genome-wide prediction of protein function is reviewed.

Abstract:Bcl-2 protein family members, including anti-apoptotic and pro-apoptotic members, act on mitochondria controlling the fate of cell between life and death. In healthy cells, Bcl-2 protein family displays specifically cellular location that fit to their functioning: anti-apoptotic members are localized predominantly to the cellular inner membranes, especially the outer mitochondrial membrane, and most of the pro-apoptotic members exist mainly in the cytosol. Following death stimuli, Bcl-2 protein family members can be regulated through mechanisms such as phosphorylation, proteolysis and protein-protein interaction etc, by which one of the main consequences is the shift of the cellular location of proapoptotic members. Translocation of proapoptotic members from cytosol to mitochondria will result in mitochondrial dysfunction and the release of apopogenic factors in the mitochondrial intermembrane space, culminating in apoptosis.

Abstract:Aggrecan degradation is an important factor in the erosion of articular cartilage in arthritis. Aggrecanase is a newly cloned enzyme which degrades the core protein of aggrecan between the Glu³⁷³ and Ala³⁷⁴, a site different from the matrix metalloproteinases cleavage site, Asn³⁴¹～Phe³⁴². The discovery of aggrecanase and searching for its inhibitors will accelerate the development of therapeutic agents for arthritis.

Abstract:B lymphocyte stimulator (BLyS) is a recently identified member of the tumor necrosis factor (TNF) superfamily that costimulates B lymphocyte proliferation and differentiation. It prominently enhances the humoral responses to both T cell-independent and T cell-dependent antigens. BLyS is required to sustain proliferation of splenic germinal center (GC) B cell in vivo. As an antiapoptotic cytokine, BLyS provides survival signals by inducing up-regulation of Bcl-2. Transgenic mice overexpressing BLyS in lymphoid cells develops symptoms characteristic of systemic lupus erythaematosus (SLE).

Abstract:To understand the control of growth and glucose repression in Saccharomyces cerevisiae by glucose transport, a set of S.cerevisiae strains with variable expression of only one glucose transporter, Hxt7, the most abundantly expressed high-affinity transporter, was constructed. The strains were constructed by partial deletion of the HXT7 promoter in vitro and integration of the gene at various copy numbers into the genome of an hxtΔ (hxt1-hxt7 gal2 deletion) strain. The 149 bp DNA region －495 to －346 in the HXT7 promoter plays an important role in HXT7 expression. In the mutant strains with promoter length of more than 495 bp, the expression of HXT7 at high glucose concentrations was much higher than that in the wildtype strain. The level was dependent on copy number and promoter length. Increased expression at low glucose was maintained in these mutants. Hxt7 in the hxt null strain displayed an incomplete glucose repression. The growth rate correlated with the level of HXT7 expression at high glucose concentrations.

Abstract:Based on circadian rhythms experimental results of the immune system, it is assumed that the adrenal cortical hormone influences the migration and distribution of recirculating T lymphocyte between the lymphoid nodes, the blood and the spleen and the effects of cortical hormone on recirculating T cells in the lymphoid nodes and the spleen are different. A mathematical model of T lymphocyte recirculation considering the role of plasma cortical hormone level is presented. The action strength of the cortical hormone, the parameter ranges and the dependence of the modeling behaviors on parameters are explored. The model can explain stable oscillations of T lymphocytes in different lymphoid tissues and the blood, the numerical results are consistent with immune circadian rhythms experiments.

Abstract:The gp37 genes of Spodoptera litura NPV (SpltMNPV) was cloned and sequenced. Employing computer analysis, glycosilated sites and the promoter motif existed in the gp37 gene. and was shown as a late gene encoding a 37 ku glycoprotein. A phylogeny tree was designed by the comparison of their homologues from known gp37 genes of baculoviruses. It was shown that the phylogeny of baculoviruses on the basis of gp37 gene was different from that based on polyhedrin genes. For example, Bombyx mori NPV (BmNPV) and Autographa californica NPV(AcMNPV) were evolved in the same branch, otherwise, they were put in two distinct branches in the tree according to the analysis of polyhedrin. This result was consistent with the baculovirus phylogeny based on EGT published previously.

Abstract:A wide variety alterations are common in brains of aged individuals. Proteomatic analysis is a new power tool to reveal such changes at protein level. The two-dimensional electrophoresis (2-DE) was employed to compare the global protein patterns between adult and aged murine brains. One hundred and fifty micrograms of murine brain protein extracted with 5 mol/L urea, 2 mol/L thiourea, 2% CHAPS plus 2% SB3-10 was run immobilized pH gradient (IPG) isoelectric focusing electrophoresis as the first dimension, and then horizontal SDS-PAGE as the second dimension. The relative standard deviations(coefficient of variation) for protein number, pI, molecular mass and volume of protein in three different experiments were 4.43%±0.25%, 8.76%±5.14%, 13.00%±4.22% and 10.84%±9.16%, respectively. Totally, 996 and 1 256 protein spots were obtained in adult and aged murine brain map, repectively, of which 8 spots increased and 20 spots decreased in quantity. Furthermore, 4 spots were missing and 14 new spots emerged in aged murine brain compared with adult mice. The differentially displayed proteins between adult and aged murine brain are useful for diagnosing brain degenerative disorders.

Abstract:Micropipette-aspiring system was selected to test the adhesion property of Wistar rat osteoblasts on polylactide (PLA) and maleic anhydride modified-polylactide (MPLA) made in our laboratory. The aims were evaluating one of the material's suitability for tissue engineering-adhesion property and the modification method. The results showed that osteoblasts had better adhesion property on PLA than on glass. The adhesion force of osteoblasts on PLA at 24 hour increased about 2 times after modification. The 24 hour adhesion force of osteoblasts on MPLA was 1.3 times more than that of 15 minute. But there was no significant difference on PLA. The research demonstrates that MPLA is a better scaffold for osteoblasts adhesion and the modification method is suitable for tissue engineering.

Abstract:Neutral proteinase was covalently attached to amine-terminaled magnetic particles and cross-linking it with glutaraldehyde. Activity of MIE arrive 45 000 U/g (magnetic particles). Optimum conditions of immobilization were studied. Stabilities to heat and preservation,operational stability of free enzyme and MIE also were compared. Some properties of MIE, such as optimum pH is 7.5, optimum temperature is 60℃ were confirmed.

Abstract:Using yeast two-hybrid system to screen the protein interacting with apoptin from human leucocyte cDNA library, four clones interacting with apoptin were identified. One of them was homologue with ABP280 (actin-binding protein), ABP280 is a dimeric actin crossing protein and plays a key role in stabilizing the membrane-cytoskeleton. Cell co-immunoprecipitation showed that apoptin could bind to ABP280 in mammalian cells. Apoptin mutants T1, T2 and T3 lack the C-terminal 11 amino acid, 33～46 amino acid and both respectively. Apoptin mutants T2 and T3 failed to interact with ABP280, which revealed that its 33～46 amino acid was pivotal for the interaction. Apoptin mutant T1 still interacted with ABP280, which revealed that its C-terminal 11 amino acid was not essential for the interaction.

Abstract:Based on the linear and reverse linear differentiation models of CD8⁺ memory T cells, mathematical models were set up respectively and the dynamics of different T subpopulations was studied. It was found that when invading antigen with optimal dose of, both models can generate memory and fit well with experimental data. Further study found that CD8⁺ T cell memory relates strongly to the persistence of antigen. Thus the contribution of antigen to maintenance of T cell memory is reconfirmed. The effect of the life-span of memory cells on immune memory was also investigated. Reverse linear differentiation model is deemed to have advantage in generation of immune response and memory.

Abstract:Although LMP1 is expressed in the majority of Nasopharyngeal carcinoma(NPC), the effect of LMP1 on cellular gene expression and its contribution to the cell growth and the development of malignancy is largely unknown.CyclinD1 expression activated by LMP1 was studied.A dual-stable LMP1 integrated NPC cell line with Tet-on regulating system, designated as Tet-on-LMP1 HNE2 was used to gain insight into the cell kinetics of the induction of cyclinD1 with Western blotting. The expression of LMP1 in Tet-on-LMP1-HNE2 was tightly regulated by tetracycline or its derivation, doxycycline. LMP1 has two essential signaling domains with the carboxy terminus, termed C-terminal activation regions1(CTAR1) and CTAR2.With cell lines stably expressed wild type LMP1 , the vector or various deletion mutants and reporter gene assay, the activation essential domains of LMP1 activation cyclinD1 was also identified. The progression of cell cycle was determined by flow cytometry and soft agar assay was done to indicate that the cyclinD1 induced by LMP1 is functional.The results indicates that LMP1 induced cyclin D1 protein expression in both dose-dependent and time-dependent manner with Western blotting analysis in Tet-on-LMP1-HEN2 cell line. Reporter gene assay revealed that wild type LMP1 also can induce cyclinD1 expression at the transcriptional level via trans-activation compared to the control(11.2 fold). LMP1 deletion mutants lacking either CTAR1 or CTAR2 or both the CTAR1 and CTAR2 deletion mutants had a decreased ability to induce cyclin D1 expression(76.4%,19.3%,17.7%).The results of flow cytometry analysis pointed to a cell cycle arrest at the G0/G1 phase compared to doxycycline negative Tet-on-LMP1-HNE2 (66.42% to 56.55%).Compared with cultured with sense PS-ODN-LMP1 (30.48%), cultured with antisense PS-ODN-LMP1 and cyclinD1 showed profound decrease in colony formation(15.21%,21.76%). This is the first report showing that cyclin D1 expression could be activated by a viral protein, LMP-1. This novel finding may thus represent a direct link between LMP1 and cell cycle regulator,CyclinD1.

Abstract:In order to clone a novel putative NPC associated gene on the smallest common deletion region of 7q32-ter.BAC clone was screened by PCR using STS D7S509 probe.The up-regulated expression of 3′ end expressed sequence tags(ESTs) localized within this smallest common deletion region were screened in NPC cell line HNE1 and NPC biopsies using EST-mediated positional candidate clone and bioinformatics.The full-length cDNA of candidated EST was cloned through cDNA clone sequencing and bioinformatics.Southern blot and methylation analysis were used to study the machanism of up-regulated expression of NAG18 in NPC. The results showed that the full-length cDNA of NAG18 is 802bp,its encoding protein is 227 amino acids. It is highly homologous to human and mouse TAXREB107 and RPL6.Loss of gene copies and aberrant methylation are not the machanism of its up-regulated expression. It can be concluded that the gene NAG18 located in this region may be a putative NPC associated gene. It is a highly conserved gene. It may participate in Tax-mediated tran-activation of transcription.

Abstract:Trichosanthin (TCS), a ribosome inactivating protein extracted from the root tuber of a traditional Chinese medicinal herb Trichosanthes kirilowii, possesses anti-tumor and anti-human immunodeficiency virus (HIV) activities. With both two-photon laser scanning microscopy and confocal laser scanning microscopy in combination with the fluorescent probe Hoechst 33342, 2′, 7′-dichlorofluorescin diacetate (DCFH-DA), Indo-1 and Fluo 3-AM, TCS-induced changes in nuclear morphology, reactive oxygen species (ROS) and intracellular calcium concentration (［Ca²⁺］_i) during the apoptosis of choriocarcinoma cells (JAR) were simultaneously observed for the first time. The results indicated that TCS-induced increase in ［Ca²⁺］_i and ROS formation were involved in apoptosis of JAR cells, and that TCS-induced ROS formation was related to TCS-evoked increase in ［Ca²⁺］_i. Further studies with confocal laser scanning microscopy revealed that TCS-evoked increase ［Ca²⁺］_i was not the main factor responsible for TCS-induced ROS formation, and that TCS might induce the production of ROS through its interaction with membrane-bound receptor.

Abstract:Human lung adenocarcinoma A549 cells and cisplatin-resistant A549/DDP cells were treated with clinically relevant doses of cisplatin (30 μmol/L) and then further cultured under the same conditions. DNAs of both cell lines were extracted and subjected separately to agarose gel electrophoresis. Results showed that DNA ladders could be seen in A549 cells cultured for 12 h, while no apoptotic character appeared in cisplatin-resistant A549/DDP cells even after being cultured for 48 hour. This difference between two cell lines was further confirmed by apoptotic peaks measured with flow cytometry. Biochemical and biophysical experiments indicated that the mitochondrial membrane potential and pH_i of cisplatin-sensitive A549 cells decreased significantly, whereas the intracellular free Ca²⁺ concentration increased greatly with the culture time. But the mitochondrial membrane potential and pH_i remained at a relatively high level and the intracellular free Ca²⁺ concentration was reduced gradually with the culture time for cisplatin-resistant A549/DDP cells. It would be suggested that the anti-apoptotic character of A549/DDP cells is related with the relative intracellular basification, the persistence of mitochondrial membrane potential and decrease in intracellular free Ca²⁺ concentration which would be responsible for the resistance of A549/DDP cells to cisplatin.

Abstract:CryIAc protein of Bacillus thuringensis expressed in E.coli transformed with pQEBt was purified by Ni-NTA affinity chromotography. One phage (PH5) that binds specifically to Bt protein was selected from a combinatorial library of random 7-mer peptides fused to minor coat protein (pⅢ) gene of the filamentous coliphage M13 through three cycles of biopanning and ELISA analysis. A new method of recombination phage aided detection of insecticidal Bt protein in transgenic plants was developed using PH5. The phages selected should also be useful in the structure study of functional domains of Bt insecticidal proteins.

Abstract:Site-specific DNA recombinant system Cre/loxP from bacteriophage P1 has been developed as a novel tool for DNA manipulation, and used successfully both in vitro and in vivo. Here a protocol for effective excision of neo gene located on chromosome of Salmonella have been developed. In order to establish a tetracycline-induced expression system in the attenuated Salmonella typhi CVD908 strain, a fused DNA fragment consisting of genes of tetracycline repressor (tetR) as well as neo gene flaked by two loxP sequences in same orientation has been integrated into a defined Δaro C locus of CVD908 strain via homologous recombination. To excise the neo gene from the locus, the suicide plasmid pJG9/Cre expressing Cre recombinase under the control of tetracycline response promoter PL_tetO-1 was constructed and electropolated into the CVD908 strain. The expression of the Cre recombinase induced by anhydrotetracycline successfully excised the neo gene. Since the suicide plasmid contains SacB gene encoding an enzyme that is lethal to G- bacteria in the presence of sucrose, growing of the bacteria in a medium containing 10% sucrose cured the pJG9/Cre plasmid. Both antibiotic and PCR identification demonstrated the successful excision.

Abstract:A procedure of differential display reverse transcription polymerase chain reaction (DDRT-PCR) applicable for silver staining was optimized by adjusting the amount of several critical reagents, including total RNA, anchor primer, arbitrary primer, cDNA and dNTP. The PCR amplification products were separated on 6% vertical denaturing polyacrylamide gels. Numerous and distinct bands could be detected by silver staining. The minimum number of bands in one lane was 40, the maximum was 80 and the average was 60. The range of displayed PCR products extended from about 100 base to about 900 base. The sensitivity was 5 pg/mm². This procedure was simple, time-saving, high sensitivity and reproducible. Based on the improved procedure, the differential gene expression were studied between immature siliques of Arabidopsis wide-type and ast mutant. From nearly 16 000 cDNA fragments analyzed, 28 differential cDNA fragments were screened. After the second PCR amplification, 13 differential cDNA fragments were identified among which 7 fragments were wild type specific and 6 fragments were ast mutant specific.

Abstract:In order to detect plant thykaloid protein phosphorylation in vivo rapidly, a new approach was introduced by using INDIA^TM phosphorylated protein probe. Eight phosphoprotein bands were detected by this method in the thylakoid membranes isolated from pea leaf discs illuminated at 400 μmol·m^-2·s^-1. The molecular masses of those phosphoproteins were 65 ku, 45 ku, 36 ku, 33 ku, 30 ku, 29 ku, 20 ku and 10 ku. Respectively by using various polyclonal antibodies, those phosphoprotein bands were identified as phosphorylated D1/ phosphorylated D2 dimer, PSⅡ core phosphoproteins, CP43 (45 ku), D2 (36 ku), D1 (33 ku) and psbH gene product (10 ku), and the light-harvesting complex (LHCⅡ) phosphopolypeptides, LHCB1 (30 ku) and LHCB2 (29 ku). A comparison was made between this new approach with other methods of detecting phosphoprotein such as radiolabeling experiment or immunological blot using phosphothreonine antibody.

Abstract:A new rolling culture system with double-mouthed roller was manufactured. Hybridoma, which produces anti-hCG monoclonal antibody (mAb), was employed in the double-mouthed roller, in which growth and antibody yield of the cells were compared with those cultivated in T flask. The results showed that with an increase in growth of the cells cultured in the roller by 1.4～1.8 folds, the yield of the antibody increased by 1.3 folds of those statically cultured in T flask. Addition of gelatin in culture media could further improve both yield of the antibody and growth of the hybridoma cells.

Abstract:Using crossed experiment method, the influence of casting solutions proportion on the cellulose acetate membrane immobilized acylase was studied, and the properties of the membrane: bubbling point pressure, aperture, porosity and rate of dankness were characterized. The results showed that when the casting solution proportion is optimum, the yield of enzymatic activity can reach 98.2% and enzyme membrane has proper rate of dankness. The residual activity can get 79.7% after it is used repeatedly for 10 times. and after being stored at 4℃ for 60 days almost no out-of-activity is observed.

Abstract:C-type lectin superfamily in snake comprises a group of proteins which display anticoagulation or various other functions. PCR primers were designed according to N-terminal sequence of a snake venom protein of which the complete amino acid sequence had been unknown. Total RNA was extracted from venom gland of Agkistrodon acutus from Hunan Province, and was reversely transcribed to cDNA. Then by applying the touchdown method to anchored-PCR, with only one specific and one universal primer, two relatively distinct bands were amplified in a single non-nested PCR reaction of no more than 30 cycles. Cloning and sequencing indicated that the sequence is homologous with other members of the C-type lectin superfamily in snake. High level fusion expression in E.coli was achieved, with the fusion protein accounting for 26%～30% of total proteins.

Abstract:In order to improve the affinity of anti-CD3 ScFv, one site-directed mutagenesis method using “megaprimer” PCR was designed. Adopting the mutagenic primer in the first round of PCR, mutation DNA fragments of 180bp and 160 bp were amplified, respectively. Using these fragments as “megaprimers” in the second round of PCR, specific full-length DNA by changing the PCR program and adjusting the concentration of the primer and template were obtained. The sequencing results showed that the mutant pool of anti-CD3 ScFv had been yielded successfully.

Abstract:Human neuronal tau-40 was acetylated and then phosphorylated by neuronal cdc2-like kinase (NCLK) with ［γ-³²P］ ATP, and then the phosphorylated acetyl-tau (PAtau) was digested with pepsin. Compared with the phosphorylated tau (Ptau), two new fractions of radioactivity from PAtau were obtained as it was eluted by C-18 column on HPLC.

Abstract:To develop a precise and rapid method for glucose-6-phosphate dehydrogenase (G6PD) assay on the auto-analysis eliminating the interruption of 6-glucose-phosphate dehydrogenase (6GPD).Human blood samples from 107 normal people and 31 patients with obvious G6PD deficiency (Methemoglobin MHb) were analyzed by Rate A with sample blank correction is desired. Results showed that the measured sensitivity was 0.27 U/gHb, the within-run precision was 0.145. the coefficient of variation (CV) was 5.6% and the between-run precision was 0.229. the CV was 9.4%.The linear was between 0～15 U/gHb.Compared to the results measure by Rate A with sample blank correction is desired. The correlation coefficient(r)to two methods was 0.863.These results indicate that it is a sepificity, stable, simple and rapid method and this method is deserved to be popularized.