Objective To study the effect of cyclosporine combined with eleven acid testosterone on serum T helper cells in patients with chronic aplastic anemia.Methods A total of 86 patients with chronic aplastic anemia received treatment in our hospital were collected as the research objects,and divided into experimental group and control group,patients in the experimental group received eleven acid testosterone combined with cyclosporine therapy,patients in the control group received eleven acid testosterone therapy.Blood routine indexes,Th1,Th2,Th17 and Treg cells contents of the two groups before and after treatment were compared.Results Before treatment,hemoglobin content,leukocyte count,platelet count,peripheral blood Th1,Th2,Th17 and Treg cells contents of the two groups had no significant difference(P>0.05).After two courses of treatment,hemoglobin content,leukocyte count,platelet count of the experimental group were significantly higher than those of the control group.After two courses and four courses of treatment,peripheral blood Th1 and Th17 contents of experimental group were significantly lower than those of the control group,Th2 and Treg contents were significantly higher than those of the control group.Conclusion Eleven acid testosterone combined with cyclosporine treatment could be more effective to stimulate hematopoiesis,regulate immune function and T lymphocyte subsets content,it′s an effective method for the treatment of chronic aplastic anemia.

Objective To investigate the effects of 2-methoxyestradiol( 2-ME) on apoptosis of K562 cells and its mechanisms. Methods The K562 cells were cultured and divided into three groups. The control group: K562 cells were cultured without 2-ME treatment. The experimental group: K562 cells were cultured in the medium containing different concentrations of 2-ME (1, 2, 4, 8 and 16 μmol/L) for 36 h. The negative control group: K562 cells were replaced by water without RNase in the medium containing different concentrations of 2-ME for 36 h. The apoptosis rate, the protein and its mRNA expression of Caspase-3 and XIAP, the activity of superoxide dismutase (SOD) and reactive oxygen species (ROS) of K562 cells wasdetected by TUNEL, flow cytometry (FCM), half-quantitative RT-PCR and xanthenes oxidized enzyme assay,respectively. Results After treated with 2-ME at different concentrations for 36 hours, in the specified concentration range, 2-ME induced apoptosis of K562 cells in a concentration-dependent manner. The possible functional mechanism of 2-ME was to up-regulate Caspase-3 but down-regulate XIAP mRNA expression, and increase ROS activity but decrease SOD activity. Conclusion 2-ME can induce apoptosis of K562 cells in a concentration-dependent manner and indicate its promising potential in the treatment of chronic myeloid leukemia(CML) patients.

Smu_195c is a protein with 86 amino acids in Streptococcus mutans, a primary pathogen for human dental caries. The specific function of Smu_195c is still unknown and there are no conserved domains in it. In order to find out its function, the gene encodes Smu_195c was cloned and expressed in E. coli as N-terminally 6*His tagged recombinant protein. Two crystal forms were obtained by the hanging drop method. Form Ⅰ belongs to space group P6122 or P6522 with the unit cell parameters a = b = 62.93 (A), c= 90.63 (A), γ=120° and form Ⅱ belongs to the space group P41212 or P43212 with the unit cell parameters a =b=57.97 (A), c = 103.51 (A).Crystals from the protein with His-tag belong to form Ⅰ, however, crystals from the protein without His-tag belong to both.

Deoxycytidylate (dCMP) deaminase is an enzyme belonged to dCMP cyt deam family. The dCMP deaminase from Streptococcus mutans UA159 was cloned and expressed in E. coli, and purified to homogeneity. The FPLC size exclusion chromatography analysis reveals that the S. mutans dCMP deaminase forms hexamer in solution. The protein was crystallized using hanging drop vapour-diffusion method and diffracted to a resolution of 3.1 (A). The diffraction data were collected at BSRF beamline3W1A. The crystals belong to P213 space group, with unit cell parameters a = b = c = 113.2(A), α =β = γ = 90°. Assuming there are two subunits per asymmetric unit, the Matthews coefficient is 3.6 (A)3 ·Da-1. This is the first crystallization report of the wild-type deoxycytidylate deaminase.

Deoxycytidylate (dCMP) deaminase is an enzyme belonged to dCMP cyt deam family. The dCMP deaminase from Streptococcus mutans UA159 was cloned and expressed in E. coli, and purified to homogeneity. The FPLC size exclusion chromatography analysis reveals that the S. mutans dCMP deaminase forms hexamer in solution. The protein was crystallized using hanging drop vapour-diffusion method and diffracted to a resolution of 3.1 ?. The diffraction data were collected at BSRF beamline 3W1A. The crystals belong to P213 space group, with unit cell parameters a = b = c = 113.2 ?, ? = ? = ? = 90?. Assuming there are two subunits per asymmetric unit, the Matthews coefficient is 3.6 ?3?Da-1. This is the first crystallization report of the wild-type deoxycytidylate deaminase.

OBJECTIVETo evaluate the feasibility of genetic and prenatal diagnosis for a family affected with pyruvate kinase deficiency (PKD).

METHODSTargeted sequence capture and high-throughput sequencing technology was used to detect the exons and exon-intron boundaries of the PKLR gene in a clinically suspected PKD patient. Meanwhile, the genotype of the pedigree was validated by Sanger sequencing. Prenatal genetic diagnosis was performed by amniotic fluid sampling after genotype of the mother of the proband was determined.

RESULTSThe proband was found to harbor double heterozygous mutations, c.661G>A (Asp221Asn) and c.1528C>T (Arg510Ter), which resulted in amino acid substitution Asp221Asn and Arg510Ter. Such mutations were confirmed by Sanger sequencing. The mother and father of the proband were detected to have respectively carried c.1528C>T (Arg510Ter) and c.661G>A (Asp221Asn) mutation. The fetus was found to have carried the same mutations as the proband. Following selected abortion, analysis of fetal tissue was consistent with the result of prenatal diagnosis.

CONCLUSIONThe compound mutations of c.661G>A and c.1528C>T of PKLR gene probably underlie the PKD in the family. Prenatal diagnosis of the mutations analysis can facilitate detection of affected fetus in time.

Adult ; Anemia, Hemolytic, Congenital Nonspherocytic ; embryology ; enzymology ; genetics ; Base Sequence ; Child, Preschool ; DNA Mutational Analysis ; Exons ; Female ; Genotype ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Pedigree ; Pregnancy ; Prenatal Diagnosis ; Pyruvate Kinase ; deficiency ; genetics ; metabolism ; Pyruvate Metabolism, Inborn Errors ; embryology ; enzymology ; genetics