The preparation and application of universal group O donor red blood cells (RBC) are a trend of future transfusion medicine. This article reviewed the technologies for producing universal RBC in recent years. One of them is modification of blood group antigens, which includes two basic methods. One of these two methods is enzymatic cleavage of the terminal immunodominant sugars from carbohydrate chains on the membrane of group A or/and group B RBC, in order to produce so-called enzyme-converted group O (ECO) RBC. ECO RBC have been produced from whole units of B RBC, which then survived normally when given to type A and O individuals in clinical trial. Because of the complexity of group A antigens, conversion of group A RBC (especially A1 RBC) to group O RBC is more difficult. Recently, a new bacterial glycosidase efficiently cleaving antigens on the surface of both A₁ and A₂ RBC has been obtained. Another method is pegylation, which camouflage the antigens on the surface of RBC with non-immunogenic molecules such as polyethylene glycol (PEG) in a non-specific way, to provide O, minor antigen negative phenotype RBC. The second technology is generating universal RBC from stem cells (such as hematopoietic stem cells, human embryonic stem cells) and human dermal fibroblasts, which will provide a new resource for blood supply. Great progress has been made, but a number of challenges still remain for using them in clinical transfusion, including scale-up, effectiveness and safety of prepared RBC. However, these researches will provide solutions for the problems in current transfusion, such as blood supply shortage, blood borne disease and emergency blood transfusion, and enhance the safety of clinical transfusions in the near future.
ABO Blood-Group System ; Cell Culture Techniques ; Embryonic Stem Cells ; Erythrocyte Count ; Erythrocyte Transfusion ; Erythrocytes ; Hematopoietic Stem Cells ; Humans

Bacillus subtilis B6-1 was used as an original strain for mutagenic treatment and a defined medium was used as the selective medium. A mutant named B. subtilis W003 was isolated after three serial ultraviolet (UV) irradiations and one diethyl sulfate (DES) treatment. The ?-PGA yield on a rotary shaker was enhanced from 10.9 g/L in parental strain to 20.5 g/L in the mutant. It was illustrated by single factor experiments that the optimal carbon and nitrogen sources were glucose and (NH4)2SO4 respectively. The optimal fermentation medium was achieved by orthogonal test. In the optimal medium, a ?-PGA yield of 45.3 g/L was obtained after 36 h cultivation.

Poly ?-glutamate is a biopolymer material that has a good application prospect.The Vitreoscilla hemoglobin(VHb) gene was integrated into the chromosome of Bacillus licheniformis WX-02 by integrative vector pDG1730-vgb.The expression of VHb was confirmed by CO-difference spectra analysis.It was shown by the results of batch cultures in a 3 L bioreactor that biomass and ?-PGA obtained in the recombinant M2 were 25.5 % and 20% higher than those of the control respectively.

Under the internal control perspective,it summarized the cases of daily bill management in the target hospital,found that the management method of the current hospital bill,bill time and the time of entering account book were inconsistent.Meanwhile bill management was not covered by the hospital information unifrom management.Through enhancing internal control and reconstructing hospital bill management system,it showed the effective fusion of financial dealing and bill management,optimized the process and method of hospital bill management.

Liver cancer is a common malignant tumor in China and a major health concern. We aimed to estimate the liver cancer incidence and mortality in China in 2010 using liver cancer data from some Chinese cancer registries and provide reference for liver cancer prevention and treatment. We collected and evaluated the incidence and mortality data of liver cancer in 2010 from 145 cancer registries, which were included in the 2013 Chinese Cancer Registry Annual Report, calculated crude, standardized, and truncated incidences and mortalities, and estimated new liver cancer cases and deaths from liver cancer throughout China and in different regions in 2010 from Chinese practical population. The estimates of new liver cancer cases and deaths were 358,840 and 312,432, respectively, in China in 2010. The crude incidence, age-standardized rate by Chinese standard population (ASR China), and age-standardized rate by world standard population (ASR world) were 27.29/100,000, 21.35/100,000, and 20.87/100,000, respectively; the crude, ASR China, and ASR world mortalities were 23.76/100,000, 18.43/100,000, and 18.04/100,000, respectively. The incidence and mortality were the highest in western regions, higher in rural areas than in urban areas, and higher in males than in females. The age-specific incidence and mortality of liver cancer showed a rapid increase from age 30 and peaked at age 80-84 or 85+. Our results indicated that the 2010 incidence and mortality of liver cancer in China, especially in undeveloped rural areas and western regions, were among high levels worldwide. The strategy for liver cancer prevention and treatment should be strengthened.
China ; epidemiology ; Female ; Humans ; Incidence ; Liver Neoplasms ; epidemiology ; mortality ; Male ; Registries ; Rural Population ; Sex Distribution ; Urban Population

This study was aimed to prepare a reconstructed B. Fragilis-derived recombinant α-galactosidase developed for human B to O blood group conversion. Based on the construction of recombinant E. Coli (DE3) which can express α-galactosidase, the inducing time and inducer concentration were optimized for high expression of α-galactosidase. Then, the expression products in supernatant were purified by cation and anion exchange column chromatography. The purified α-galactosidase was used to treat B group red blood cells in phosphate buffer (pH 6.8) for 2 hours to prepare O group red blood cells. The results showed that the optimal inducing conditions for α-galactosidase expression were IPTG 0.1 mmol/L, 37°C and 2 hours. The specific enzyme activity of purified protein increased from 0.42 U/mg to 2.1 U/mg as compared with pre-purification. And, the conditions of B to O blood group conversion were 26°C, pH 6.8 (neutral pH condition) and 2 hours. Moreover, 225 µg of the enzyme could converse 1 ml B red blood cells to O completely. It is concluded that the technology of expression and purification of recombinant α-galactosidase has been established, and the purified protein can converse B red blood cells to O completely, which means that an effective enzyme conversing B red blood cells to O has been obtained.
ABO Blood-Group System ; immunology ; Bacteroides fragilis ; enzymology ; Cloning, Molecular ; Escherichia coli ; metabolism ; Humans ; Recombinant Proteins ; biosynthesis ; alpha-Galactosidase ; biosynthesis

αGal, a xenotransplantations antigen (XTA), can lead to hyper acute reaction (HAR) in xenotransplantation. α-Galactosidase from B. fragilis is a novel galactosidase belong to CAZy GH110 which can clear the terminal αGal from branched and linear oligosaccharides. This study was purposed to investigate the removal effect of a novel α-galactosidase on α-Gal XTA on surface of red blood cells. The αGal XTA from the red blood cells of cattle, pig, dog and rabbit was digested by using recombinant α-galactosidase; the α-Gal antigens on surface of cells was detected by flow cytometry. The results showed that the XTA was disappeared completely or mainly. It is concluded that the novel α-galactosidase is a potential enzyme to remove the XTA on the surface of xenotransplants and can be used to overcome the HAR in xenotransplantation.
Animals ; Antigens, Heterophile ; immunology ; Cattle ; Dogs ; Epitopes ; Erythrocytes ; immunology ; Macaca mulatta ; Mice ; Mice, Inbred BALB C ; Rabbits ; Swine ; Transplantation, Heterologous ; alpha-Galactosidase ; immunology

OBJECTIVETo select short tandem repeats(STR) from X chromosome.

METHODSSTR is a universal genetic marker that has changeable polymorphism and stable heredity in human genome. It is a specific DNA segment composed of 2-6 base pairs as its core sequence. It is an ideal DNA marker used in linkage analysis and gene mapping. In this study, 8 short tandem repeats were selected from two genomic clones on X chromosome by using BCM Search Launcher. Primers amplifying the STR loci were designed by using Primer 3.0 according to the unique sequence flanking the STRs. Polymorphisms of the short tandem repeats in Chinese population were evaluated by PCR amplification and PAGE.

RESULTSFive of these STRs were polymorphic. Chi-square test indicated that the distribution of genotypes agreed with Hardy-Weinberg equilibrium (P>0.05).

CONCLUSIONFive polymorphic short tandem repeats have been identified on chromosome X and will be useful for linkage analysis and gene mapping.

Chromosomes, Human, X ; genetics ; Female ; Genotype ; Humans ; Microsatellite Repeats ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics

OBJECTIVESmith-Fineman-Myers syndrome (SFMS) is an X-linked mental retardation syndrome. The authors had ascertained a large Chinese family with SFMS from Shandong and had mapped the disease locus to an interval of 19.8 Mb on Xq25 flanked by markers DXS8064 and DXS8050. Further investigation suggested that SFMS exhibited locus heterogeneity. In this study for facilitating the identification of the gene responsible for SFMS, the additional markers were analyzed to narrow down the candidate region, and four candidate genes (GPC3, MST4,GPCR2 and GLUD2) were chosen and screened for disease-causing mutation.

METHODSPCR and denaturing polyacrylamide gel electrophoresis were used to genotype 13 new polymorphic markers distributed within the candidate region. Mutation detection was accomplished by sequencing the exons and intron-exon junctions of the candidate genes.

RESULTSBy analyzing 13 additional polymorphic markers, SFMS candidate region can be reduced to an interval of 10.18 Mb bounded by XSTR3 and XSTR4, and no disease-causing mutation was identified in the coding regions of four candidate genes.

CONCLUSIONGPCR2 GPC3, MST4 and GLUD2 were excluded as pathogenic genes for SFMS. The refined SFMS locus will assist in the identification and characterization of other candidate genes for SFMS.

Abnormalities, Multiple ; genetics ; Chromosome Mapping ; Chromosomes, Human, X ; Genetic Linkage ; Glutamate Dehydrogenase ; genetics ; Glypicans ; Humans ; Intellectual Disability ; genetics ; Male ; Membrane Proteins ; genetics ; Neoplasm Proteins ; genetics ; Protein-Serine-Threonine Kinases ; genetics ; Receptors, G-Protein-Coupled ; genetics ; Syndrome

In order to study the possibility of xenotransfusion from porcine red blood cell (pRBC) to primate, the antigens on pRBC surface were modified to make it more compatible to primate sera. Porcine RBCs were subjected to both enzymatic removal of membrane alpha-Gal antigens with recombinant alpha-galactosidase (AGL) and covalent attachment of succinimid propionate-linked methoxypolyethyleneglycol (mPEG-SPA) to camouflage non-alphaGal antigens. The effects of double modifications were determinated by hemagglutination and clinical cross-match testing with rhesus sera. In vivo clearance rates and safety of modified pRBCs were measured after it was transfused into Rhesus monkey with or without immunosuppressant treatment. The validity of pRBC was detected in exsanguine Rhesus monkey model. The results showed that AGL could effectively remove alpha-Gal xenoantigens on pRBC membrane and reduce hemagglutination. The combination of mPEG modification with AGL treatment could significantly increased compatibility between pRBCs and Rhesus monkey sera. Modified pRBCs were detectable in Rhesus monkey blood at 12 hours after transfusion, and their survival time was 40 hours in the immunosuppressant-treated Rhesus monkey. In vivo survival rates of pRBCs were 38% in exsanguine Rhesus monkey at 8 hours after transfusion, and during that time, the hemoglobin and hematocrit of Rhesus monkey were maintained at the same level as before it lost blood. It is concluded that the modified pRBC can be safely transfused into Rhesus monkey and relieve the anemic symptom exsanguine Rhesus monkey. It suggested that pRBC can be hopefully used as a blood substitute for primate and human in the future.
Animals ; Erythrocyte Transfusion ; methods ; Erythrocytes ; drug effects ; immunology ; Hemagglutination Tests ; Macaca mulatta ; immunology ; Polyethylene Glycols ; pharmacology ; Swine ; blood ; Transplantation, Heterologous ; methods ; alpha-Galactosidase ; pharmacology