Objective: To isolate the Guiqi (Angelicae Sinensis Radix and Astragali Radix) polysaccharides (AAP) and study the structural characterization of AAP-2A. Methods: Hot water extracting and ethanol precipitating method was employed to isolate the AAP. A new fraction (AAP-2A) of AAP was obtained after AAP was purified by deproteination, gel chromatography on DEAE-52 cellulose, gel chromatography on Sephadex G-100, and other means. Sugar content of AAP-2A was determined using phenol sulfuric acid method. Chromatography techniques were used to determine the ultraviolet properties, infrared properties, relative molecular weight, absolute molecular weight, molecular conformation and molecular weight distribution, and monosaccharide composition with the molar ratio of AAP-2A. The connection, main chain and branched-chain structures, and the condition of branching point were studied by methylation and NMR methods. Results: The total sugar content of AAP-2A, which was absent of proteins and nucleic acids, was 98.98%. Infrared spectroscopy showed that AAP-2A had the typical signals of polysaccharides. The relative and absolute molecular weights of AAP-2A were above 6.68 × 105 and 2.252 × 106; AAP-2A was a neutral sugar and composed of rhamnose (Rha), galactose (Gal), arabinose (Ara), and glucose (Glc) with a molar ratio of 1 : 2.13 : 3.22 : 6.18. AAP-2A proved to be a heteropolysaccharide, with 1, 3-α-L-Rha, 1,3-β-D-Gal, 1,3-, 1,5-, and 1,3,5-α-L-Ara, 1,4- and 1,4,6-α-D-Glc residues in backbone and 1-α-D-Glcp residues in branches. Conclusion: AAP-2A is a new neutral heteropolysaccharide from Guiqi polysaccharides with a highly branched conformation.

The aim of study was to investigate the inhibitory effect of small interfering RNA on evi1 gene expression and biological characteristics in HEL cells and its mechanism. 3 siRNA (siRNA-1, siRNA-2, siRNA-3) specific for evi1 gene were synthesized and transfected into HEL cells in vitro. Experiments were divided into test and control groups. MTT method was used to assay the inhibitory effect of siRNA on cell proliferation; semiquantitative RT-PCR was used to detect the expression of evi1 gene mRNA; the cell viability was determined by trypan blue dye test; the change of cell cycle and apoptosis of cells were analyzed by flow cytometry. The results showed that siRNA-1 had strongest effect, and inhibitory effect was most obvious at 48 hours after transfection. When the concentration of siRNA raised to 120 nmol/L, the inhibitory rate reached to the peak. The inhibitory rate of siRNA-1 on proliferation of HEL cells, relative expression level of evi1 mRNA and cell viability at 48 hours after transfection were 72.22 ± 2.80%, 27.31 ± 1.11% and 26.05 ± 2.49%, which had significant difference from other groups (p < 0.001). The siRNA resulted in arrest of cell cycle at G(0)/G(1) phase, the cell amount at S phase obviously decreased, the apoptotic rate of HEL cells obviously increased (p < 0.01). It is concluded that the siRNA specific for evi1 gene can suppress the proliferation of HEL cells, reduce the expression of evi1 mRNA, decrease the cell viability, arrest the cell cycle at G(0)/G(1) phase, suppress cell mitosis, and promote cell apoptosis.
Apoptosis ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; DNA-Binding Proteins ; metabolism ; Gene Expression Regulation, Leukemic ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; pathology ; MDS1 and EVI1 Complex Locus Protein ; Proto-Oncogenes ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Transcription Factors ; metabolism

AIMTo establish a simple and effective method for RBCs labeling and survival assays, and the qualities of rabbit RBCs preserved in GMA solution at 4 degrees C were verified.

METHODSThe bloods were taken through the ear arteries of the rabbits. The RBCs were labeled by fluorescein isothiocyanate (FITC), and were reinjected to the same rabbit through ear veins. The percentage of FITC labeled RBCs was assayed by FACS at a series of times after injection. The SAS software was employed to analyze the data and establish the regression equations. The 24-hour recovery and the half-life span of the labeled RBCs were calculated according to the equations.

RESULTSThe 24-hour recovery and the half-life span of the labeled RBCs in the control group were 93.76% +/- 5.40% and 22.50% +/- 4.37 days respectively, which was in agreement with the previous papers. The 24-hour recovery and the half-life span of the labeled RBCs in the GMA group were 89.13% +/- 7.10% and 11.41% +/- 1.63 days respectively, which was coincident with the infusion conditions.

CONCLUSIONCompared with other methods of RBCs labeling in vivo, FITC labeling was thought to be easier and cheaper to use, which could facilitate the analysis of the biological character of the labeled cells, and could be used to trace the fate of labeled cells.

Animals ; Blood Preservation ; methods ; Erythrocyte Aging ; physiology ; Erythrocyte Count ; Erythrocytes ; physiology ; Fluorescein-5-isothiocyanate ; Rabbits ; Software

AIMTo convenience of the methods for assaying red blood cell glycolysis without oxygen condition in the studies.

METHODSReagent kit of glucose, perchloric acid, visible light prismatic photometer, battle of nitrogen and rocking bed are used in the studies. The process includes 4 steps prepare Tris- HCI solution and so on, assay of red blood cell glycolysis without oxygen condition and account of glycolysis rate.

RESULTSHuman red blood cells stored at 4 degrees C for 75 d, in SOD solution, the glycolysis rate is 86.2% +/- 5.0%, distinctly better than GMA solution (39.2% +/- 8.9%).

CONCLUSIONThe methods of assaying glycolysis without oxygen condition not use Habea's apparatus. The operation is convenient and simple and its determinations can be performed in ordinary laboratory and is is accurate.

Erythrocytes ; metabolism ; physiology ; Glycolysis ; physiology ; Hematologic Tests ; methods ; Humans ; Oxygen ; metabolism

To study the dynamic changes of ultrastructure of erythrocytes in prolonged preservation of blood with preservative fluid containing superoxide dismutase (SOD), the whole blood samples were preserved at 4 degrees C in SOD-containing solution, the morphologic changes of erythrocyte were dynamically ob served by transmission microscopy after preservation for 42, 75 and 85 days, an d the blood samples preserved in GMA solution served as control. Three variance was applied to analyze the data with SAS software. The results showed that the metamorphotic rates of erythrocyte preserved in SOD-containing solution for 42, 75 and 85 days were lower than those of erythrocytes preserved in GMA solution. Most of metamorphotic rates of erythrocyte preserved in SOD-containing solution for 42, 75 and 85 days were correspond to those of erythrocytes preserved in GMA solution for 42 days, or even lower. It is concluded that SOD-containing preservative fluid might help to maintain the normal morphology of erythrocytes in prolonged preservation at 4 degrees C.
Blood Preservation ; Erythrocyte Deformability ; Erythrocytes ; ultrastructure ; Humans ; Microscopy, Electron ; Superoxide Dismutase ; pharmacology ; Time Factors

To study effect of pre-freezing temperature and lyophilizer shelf temperature on recovery of human red blood cells after lyophilization and determine solidifying temperature of this lyophilization system, the protective solution composed of 7% DMSO, 40% polyvinylpyrrolidone (PVP) and isotonic buffer were adopted to lyophilize red blood cells at different pre-freezing temperatures or shelf temperatures. At first, fresh whole blood was centrifugated, washed and equilibrized to prepare concentrated red blood cells. Then concentrated red blood cells were mixed with the protective solution at 1:3 and pre-freezed at different temperature (-20, -35, -45, -80 or -196 degrees C) before lyophilization in lyophilizer. To study effect of shelf temperature on lyophilization of red blood cells, red blood cells were lyophilized at different shelf temperature after pre-freeze at -80 degrees C. After lyophilization, the samples were quickly rehydrated by 37 degrees C rehydration solution. The results showed the recovery rate of red blood cells and hemoglobin after pre-freeze at different temperature and lyophilization were > 85% and > 75%, there was not significant difference among these groups, but the concentration of free hemoglobin in -196 degrees C group was significantly higher than that in other groups (P < 0.01). With decreasing of shelf temperature, the lyophilizing time was also prolonged. When shelf temperature was > or = -25 degrees C, samples were not fully lyophilized; when shelf temperature was < or = -30 degrees C, the recovery rate of red blood cells and hemoglobin after lyophilization and rehydration were above 90%; after washed to isotonic state, the recovery rate of hemoglobin of the four groups was similar to each other. In conclusion, only when pre-freezing temperature is between -20 and -80 degrees C and the lyophilizer shelf temperature is < or = -30 degrees C, the effect of lyophilization is better, but the effect of excessively low pre-freezing temperature may even be worse.
Blood Preservation ; Erythrocytes ; cytology ; Freeze Drying ; Hemoglobins ; analysis ; Humans ; Temperature

OBJECTIVETo understand the prevalence and control of several common chronic disease in Beijing adults.

METHODS16,658 adult residents were randomly selected with stratified multi-stage cluster sampling method. Each participant was invited to receive a set of standardized questionnaire, physical examinations and laboratory tests.

RESULTSData showed that the prevalence, awareness, treatment and the rate of control on hypertension among the adults in Beijing were 29.1%, 49.3%, 42.3% and 10.6% respectively. The counterparts of diabetes mellitus were 8.8%, 56.7%, 50.0% and 15.0%. The four corresponding figures for dyslipidemia were 33.2%, 31.1%, 13.0% and 4.3%, respectively. 22.9% of the Beijing adults had metabolic syndrome including 8.1 per thousand suffering from myocardial infarction and 18.4 per thousand from stroke. Except for diabetes, all the chronic diseases had higher prevalence rate in rural area than in urban area, according to the findings under our study. Postmenopausal women were more susceptible to chronic disease than men.

CONCLUSIONThe prevalence rate of chronic disease in Beijing was still high. The prevalence rate in rural area had exceeded the level in urban area. Adjustment and attention should be made according to the prevalence features and weakness existed in present chronic disease control strategy.

Adult ; Aged ; Aged, 80 and over ; China ; epidemiology ; Chronic Disease ; epidemiology ; Diabetes Mellitus ; epidemiology ; Dyslipidemias ; epidemiology ; Female ; Humans ; Hypertension ; epidemiology ; Male ; Metabolic Syndrome ; epidemiology ; Middle Aged ; Prevalence ; Young Adult

OBJECTIVETo investigate the effect of immature dendritic cells (inDC) genetically modified to express sTNFR I on acute graft-versus-host disease (aGVHD) and the graft-versus-leukemia (GVL) effect ofter allogeneic bone marrow transplantation (allo-BMT) in leukemic mice and its mechanism.

METHODSAn EL4 leukemia allo-BMT model was established with the BALB/c (H-2d) donor mice (DM)and C57BL/6 (H-2b) recipient mice (RM). The RM received DM bone marrow (BM) cells at a 1:1 ratio with spleen cells intravenously via tail vein at 4 h after TBI. Fifty DM were separated randomly into five groups: (1) Group A: total body irradiation (TBI) group, (2) Group B: lymphoma cell leukemia group, (3) Group C: allo-BMT group, (4) Group D: pXZ9-DC group, (5) Group E: sTNFR I-DC group. Acute GVHD scores, incidence of leukemic cell infiltration, histopathological analysis, survival rate, and survival rate of the recipients were estimated after allo-BMT. Enzyme-linked immunosorbent assay (ELISA) method was used to detect cytokines (INF-gamma and IL-4 ) production. Flow cytometry (FCM) analysis was used to detect allogeneic chimerism.

RESULTS(1) The mice in group A and group B all died of the BM failure and lymphoma cell leukemia, respectively. The mice in group C developed typical clinical signs of a GVHD after BMT with an average survival time(AST) of (11.50 +/- 3.50) d. The signs of aGVHD were less evident in the group D and E, and their AST (21.70 +/- 5.80 and 25.80 +/- 5.20 days, respectively) were all longer than that in group C (P < 0.05). AST of group E was the longest (P < 0.05). The mice in group B all died of leukemia within 18 days after engraftment of EL4 cells. There was was no significant difference in groups C, D and E in the incidence of leukemia (P > 0.05). (2) Serum IFN-gamma level reached peak value. At + 12 d, then decreased gradually in group C, D, and E, and then reached the nadir at +18 d post-BMT, with the lowest in group E (P < 0.05), and the level was significantly lower in group D than in group C (P < 0.05). After BMT, serum IL-4 level slightly decreased in group C, but gradually elevated in group D and E and reached their peak at +12 d, and even more significantly increased in group E (P < 0.05). There was no statistical significance in the pair wise comparison among three group (P < 0.05). (3) The average proportion of H-2d positive cells in RM was 95%-100% on day 30 post-BMT, with complete donor-type implantation.

CONCLUSIONImmature DC can induce immuno tolerance. Immature DC genetically modified to express sTNFR I has been shown to prevent acute GVHD in lethally irradiated mice reconstituted with allogeneic bone marrow grafts while maintaining the GVL response.

Animals ; Bone Marrow Transplantation ; adverse effects ; methods ; Dendritic Cells ; immunology ; Female ; Graft vs Host Disease ; prevention & control ; Graft vs Leukemia Effect ; Immune Tolerance ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Receptors, Tumor Necrosis Factor, Type I ; genetics ; Transplantation, Homologous

OBJECTIVETo develop a general quality of life (QOL) instrument for Chinese in accordance with the Chinese culture and to assess its reliability, validity and sensitivity.

METHODSA 35-item QOL questionnaire(QOL-35) was developed with reference to the World Health Organization QOL questionnaire(WHO-100) and the 36-item medical outcomes study on short-form health status(SF-36). Thirty five items were divided into six domains (general, physical, independent, psychological, social, environment) and one item on QOL transition. The reliability of QOL-35 was assessed by a test-retest survey among 127 adults with an interval of 24-72 hours. The internal consistency and validity were evaluated by a survey on 135 adults from outpatients or general population, using QOL-35, WHO-100 and SF-36. The adaptability was assessed by application to 1356 community-based samples in Beijing.

RESULTS(1)Test-retest reliability of QOL-35: weighted Kappa indexes for items were from 0.86 to 1.00. Intraclass correlation coefficients were from 0.68 to 0.94 for domains, and 0.94 for total score. (2) On internal consistency: Cronbach's Alphas were 0.93, 0.97 and 0.89 for QO1-35, WHO-100 and SF-36. (3)On construct validity. The accumulated proportions of variances of the preceding seven factors were 66.5%, 50.3% and 65.3% for QOL-35, WHO-100 and SF-36. (4) On criterion validity. Spearman correlation coefficients of total QOL score of QOL-35 with those of WHO-100 and SF-36 were 0.805 and 0.745. (5)The rates of chronic diseases were 53.1%, 33.1%, 26.4% and 25.1% from first to fourth quantile of the total QOL scores of QOL-35(P<0.05). (6)Cronbach's Alpha was from 0.68 to 0.93 in 135 subjects, and from 0.71 to 0.91 in 1356 individuals of natural population.

CONCLUSIONThe QOL-35 instrument satisfied test-retest reliability and was highly correlated with WHO-100 and SF-36, having fewer items but better construction validity, better internal consistency, and better discrimination ability. We suggested that QOL-35 be used as a replicable tool to assess quality of life in the Chinese general population.

Adult ; Aged ; Aged, 80 and over ; Analysis of Variance ; China ; ethnology ; Chronic Disease ; Female ; Humans ; Male ; Middle Aged ; Quality of Life ; Reproducibility of Results ; Surveys and Questionnaires ; World Health Organization

AIMTo investigate the mechanism of protective effect of SOD (superoxide dismutase) on damage of RBCs stored at 4 degrees C, the studies of erythrocyte glucose and energy metabolism were performed.

METHODSwhole blood collected from healthy donors and stored at 4 degrees C in ACD, GMA and SOD solutions. Before and post storage, some parameters were assayed. Standard methods were used for the in vitro tests. The 24-hour in vivo recoveries were measured by FTTC (Fluorescein 5-isothiocyanate) from SIGMA Company.

RESULTSAll parameters of red blood cell glucolysis rate without oxygen condition, ATP, PK (pyruvic kinase) and 24 h recoveries level were 86.2%, 56.4%, 64.3% and 86.2% of normal respectively stored in SOD solution at 4 degrees C for 75 days, distinctly more than in ACD and GMA groups at 75 days stored. The 24 h recovery at 75d in group SOD was near the recovery at 42d in group GMA.

CONCLUSIONWhole blood in SOD solution can be stored satisfactorily for 75 days at 4 degrees C, and furnished theoretical evidence for RBCs survival.

Animals ; Blood Preservation ; methods ; Erythrocytes ; cytology ; metabolism ; Humans ; Rabbits ; Refrigeration ; Superoxide Dismutase ; pharmacology