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

The study was aimed to investigate the possibility of enhancing transfection efficiency of branched polyethylenimine (BPEI) in HeLa cells by hydrophobic tail of bee venom peptide (melittin). Hydrophobic tail of melittin was synthesized and its membrane permeable activity was evaluated by hemolysis test. The peptide was mixed with BPEI and the transfection efficiency was determined in HeLa cells by using green fluorescent protein gene (GFP) as a reporter gene. The cytotoxicity of the mixture was analyzed by MTT assay at 24 hours after transfection. The results indicated that the synthesized peptide had permeable activity leading to hemolysis in both neutral and acidic solution. At optimal condition, the peptide could significantly improve the transfection efficiency of BPEI and the cytotoxicity of the mixture was lower than BPEI itself. It is concluded that hydrophobic tail of melittin may be a potential enhancer to improve transfection efficiency mediated by cationic polymers in difficult to transfect cells.
HeLa Cells ; Humans ; Hydrophobic and Hydrophilic Interactions ; Melitten ; chemistry ; genetics ; Peptides ; chemistry ; Polyethyleneimine ; pharmacology ; Transfection

To investigate the suitable site environment for Atractylodes lancea, field trials in different niches was carried out, and the seedling growth, biomass and volatile oil components in different microhabitas was studied. The study found that the survival rate, plant height, volatile oil content of those which growing under the bamboo were significantly higher than those exposed in the open field. The survival rate understory was (76 ± 15.1) %, plant height understory was (77 ± 14. 8) cm and the summation of the four kinds of volatile oil content understory reached up to 4.09%; The same evaluation values of these indicators of the four faces in the open field respectively: survival rate is 30%, plant height was (77 ± 14.8) cm and the summation of the four kinds of volatile oil content was 2.24%. But, the yield of the understory (41 ± 22.3) g was significantly lower than those four faces in the open field (104.5 g) on the contrary. For the four open field towards, the yield of the east facing, which organic matter and other nutritional conditions were better than others, was significantly higher than those in the other facings. A. lancea was found to be an anti-poor and shading-like or growing in east facing slope herb through the correlation analysis of light, temperature, soil and leaf nutrients with seedling growth, biomass and volatile oil components. It also reminds us that the understory herb with high survival rate, low incidence, low management costs, and high medicinal ingredients, although it's production is not so high, but it can be improved by increased organic fertilizer. So the ecological planting patterns which can intercropping herbs with the forest was proposed.
Atractylodes ; chemistry ; growth & development ; metabolism ; Drugs, Chinese Herbal ; chemistry ; metabolism ; Molecular Structure ; Oils, Volatile ; chemistry ; metabolism ; Seedlings ; chemistry ; growth & development ; metabolism ; Soil ; chemistry ; Temperature

BACKGROUND: Ischemia-reperfusion injury in the myocardium after cardiac arrest (CA) and cardiopulmonary resuscitation (CPR) is an important pathologic basis of post-cardiac arrest of syndrome (PCAS), and apoptosis is one of the major mechanisms in myocardial ischemia-reperfusion injury. To lessen myocardial ischemia-reperfusion injury after cardiac arrest and CPR, it is important to reduce energy consumption and to increase energy supply in the myocardium. This study aimed to observe changes of cell apoptosis and expression of Bcl-2 and Bax protein on the myocardium after CPR in rats, and the protective effects of different doses of exogenous phosphocreatine (creatine phosphate, CP) on them. METHODS: A total of 32 male adult Sprague-Dawley rats were randomly divided into 4 groups: control group (group A), CPR group (group B), low-dose CP group (group C, CP 0.5 g/kg at beginning of CPR and 1.0 g/kg at 2 hours after CPR) and high-dose CP group (group D, CP 1.0 g/kg at beginning of CPR and 2.0 g/kg at 2 hours after CPR). Cardiac arrest was induced by asphyxiation and CPR started at 7 minutes after asphyxiation in groups B, C and D. Myocardium samples were taken at 24 hours after CPR. Cardiomycytic apoptosis was detected by the TdT-mediated dUTP-biotin nick end labeling (TUNEL) method. The expression of Bcl-2 and Bax protein was measured by immunohistochemistry. RESULTS: Cardiomyocytic apoptosis index (AI) and expression of Bcl-2 and Bax protein increased more significantly in groups B, C and D than in group A (P<0.01), but Bcl-2/Baxratio significantly decreased (P<0.01). Cardiomyocytic AI and expression of Bcl-2 and Bax protein decreased more significantly in groups C and D than in group B (P<0.01), but Bcl-2/Bax ratio increased more significantly (P<0.01). Cardiomyocytic AI and expression of Bcl-2 and Bax protein decreased more significantly in group D than in group C (P<0.05), but Bcl-2/Bax ratio increased more significantly (P<0.05). CONCLUSION: Exogenous phosphocreatine, especially at a large dose, could inhibit cardiomyocytic apoptosis and alleviate myocardial injury after CPR in rats.

20?10~9/L.The proteinuria decreased or disappeared.The antinuclear antibody decreased or became negative.The level of complement was increased.The following complications were ob- served:septicemia in 2 patients,cytomegalovirus infection in 2 patients,renal toxicity in 1 patient,acute left heart failure in 3 patients and cardiac arrhythmia in 3 patients.There was no transplantation related mortality. Conclusion APBSCT may improve the disease activity and the immunological markers in SLE.It is a valid therapy for refractory SLE,but the long-term effects need to be observed.

Objective To develop the method of 16S rRNA gene clone library for tick bacterial flora analysis, and to analyze the detection effective of pathogens in tick and capacity of bacterial flora diversity. Methods Primers were designed according to the specific gene of Borrelia burgdorferi, Bartonella henselae, Anaplasma phagocytophilum, Ehrlichia chaffeensis and templates were choosen by positive PCR result to amplify the DNA extracted from the ticks. One set of primers targeting 16S rRNA gene conserved region were chosen to amplify certain fragments, DNA extraction, PCR reaction, cloning and sequencing. Nucleotide sequences were compared with GenBank database. Calculated Coverage values of clone library and Shannon-Wiener diversity index. Results Sixteen defined genus-or species-bacteria were detected in 103 valid sequences. Eight species were edge type (Clone No. > 5). Three kinds of pathogens were identified (Borrelia burgdorferi, Bartonella henselae and Rickettsia sp). Three kinds of pathogens were not edge type(Clone No. < 5). Coverage value was 96.11%, and Shannon-Wiener index was 2.40. Analysis results of cloning sequence showed that tick-parasitic bacteria mainly were α and γ deformation mycetes which accounted for 56.25% (9/16). Conclusions The 16S rRNA gene sequences technology could make relative quantitative of bacterial flora, and detect many kinds of pathogens in tick. It's a good method for detection of pathogens and bacterial flora analysis.

OBJECTIVE: To evaluate the role of contrast enhanced power Doppler in evaluating tumor angiogenetic activity. METHODS: Thirty-two patients with hepatocellular carcinoma were analysed. Flow signals of hepatocellular carcinoma were observed by power Doppler imaging after the injection of contrast agent, and then the relative perfusion rate and blood flow were assessed. The microvessel density (MVD) and vascular endothelial growth factor(VEGF) were assessed by immunohistochemical method. The relationship between the relative perfusion rate,blood flow, MVD,VEGF was studied. RESULTS: The relative perfusion rate in the tissues with positive expression of VEGF was significantly higher than that in the tissues with negative expression of VEGF in hepatocellular carcinoma. There was correlation between the relative perfusion rate, blood flow grade and MVD(P<0.05). The expression of VEGF was positively related to the relative perfusion rate and blood flow grade(P<0.05). CONCLUSION Contrast enhanced power Doppler is useful in evaluating the tumor angiogenetic activity.
Aged ; Carcinoma, Hepatocellular ; blood supply ; diagnostic imaging ; Contrast Media ; Female ; Humans ; Liver Neoplasms ; blood supply ; diagnostic imaging ; Male ; Middle Aged ; Neovascularization, Pathologic ; Ultrasonography, Doppler ; methods

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

BACKGROUND:The outcome of cardiopulmonary resuscitation (CPR) may depend on a variety of factors related to patient status or resuscitation management. To evaluate the factors influencing the outcome of CPR after cardiac arrest (CA) will be conducive to improve the effectiveness of resuscitation. Therefore, a study was designed to assess these factors in the emergency department (ED) of a city hospital.METHODS:A CPR registry conforming to the Utstein-style template was conducted in the ED of the First Affiliated Hospital of Wenzhou Medical College from January 2005 to December 2011. The outcomes of CPR were compared in various factors groups. The primary outcomes were rated to return of spontaneous circulation (ROSC), 24-hour survival, survival to discharge and discharge with favorable neurological outcomes. Univariate analysis and multivariable logistic regression analysis were performed to evaluate factors associated with survival.RESULTS:A total of 725 patients were analyzed in the study. Of these patients, 187 (25.8％) had ROSC, 100 (13.8％) survived for 24 hours, 48 (6.6％) survived to discharge, and 23 (3.2％) survived to discharge with favorable neurologic outcomes. A logistic regression analysis demonstrated that the independent predictors of ROSC included traumatic etiology, first monitored rhythms, CPR duration, and total adrenaline dose. The independent predictors of 24-hour survival included traumatic etiology, cardiac etiology, first monitored rhythm and CPR duration. Previous status, cardiac etiology, first monitored rhythms and CPR duration were included in independent predictors of survival to discharge and neurologically favorable survival to discharge.CONCLUSIONS:Shockable rhythms, CPR duration ≤15 minutes and total adrenaline dose ≤5 mg were favorable predictors of ROSC, whereas traumatic etiology was unfavorable. Cardiac etiology, shockable rhythms and CPR duration ≤15 minutes were favorable predictors of 24-hour survival, whereas traumatic etiology was unfavorable. Cardiac etiology, shockable rhythms, CPR duration ≤15 minutes were favorable predictors of survival to discharge and neurologically favorable survival to discharge, but previous terminal illness or multiple organ failure (MOF) was unfavorable.

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