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Red blood cells are destroyed when the shear stress in the blood pump exceeds a threshold, which in turn triggers hemolysis in the patient. The impeller design of centrifugal blood pumps significantly influences the hydraulic characteristics and hemolytic properties of these devices. Based on this premise, the present study employs a multiphase flow approach to numerically simulate centrifugal blood pumps, investigating the performance of pumps with varying numbers of blades and blade deflection angles. This analysis encompassed the examination of flow field characteristics, hydraulic performance, and hemolytic potential. Numerical results indicated that the concentration of red blood cells and elevated shear stresses primarily occurred at the impeller and volute tongue, which drastically increased the risk of hemolysis in these areas. It was found that increasing the number of blades within a certain range enhanced the hydraulic performance of the pump but also raised the potential for hemolysis. Moreover, augmenting the blade deflection angle could improve the hemolytic performance, particularly in pumps with a higher number of blades. The findings from this study can provide valuable insights for the structural improvement and performance enhancement of centrifugal blood pumps.

Interventional micro-axial flow blood pump is widely used as an effective treatment for patients with cardiogenic shock. Hemolysis and coagulation are vital concerns in the clinical application of interventional micro-axial flow pumps. This paper reviewed hemolysis and coagulation models for micro-axial flow blood pumps. Firstly, the structural characteristics of commercial interventional micro-axial flow blood pumps and issues related to clinical applications were introduced. Then the basic mechanisms of hemolysis and coagulation were used to study the factors affecting erythrocyte damage and platelet activation in interventional micro-axial flow blood pumps, focusing on the current models of hemolysis and coagulation on different scales (macroscopic, mesoscopic, and microscopic). Since models at different scales have different perspectives on the study of hemolysis and coagulation, a comprehensive analysis combined with multi-scale models is required to fully consider the influence of complex factors of interventional pumps on hemolysis and coagulation.

There are 2 techniques for detecting red blood cell survival (RBCS) detection techniques: red blood cell labeling test and carbon monoxide (CO) breath test. The former has disadvantages such as long measurement times and complicated procedures, while the latter is simple, convenient, moderately priced, and capable of dynamically monitoring changes in RBCS before and after treatment. Currently, the CO breath test is gradually being implemented in clinical practice. RBCS is not only applied to hematologic diseases such as multiple myeloma, myelodysplastic syndromes, lymphoma, and thalassemia, but also to non-hematologic diseases like type 2 diabetes and chronic kidney disease. It can assist in diagnosis, guide treatment, evaluate drug treatment efficacy, and predict disease progression.

Objective: To investigate the feasibility of cationic antimicrobial peptide cathelicidin-PY(PY) therapy through a mouse model of acute liver failure. Methods: The ability of different concentrations of antimicrobial peptide PY to neutralize endotoxin / lipopolysaccharide (LPS) in vitro was detected by Limulus Amebocyte Lysate (LAL) assay. Cell counting kit-8 (CCK-8) was used to detect the toxic effect of different concentrations of antimicrobial peptide PY on mouse monocyte macrophages (RAW264.7). An in vitro hemolysis experiment was used to evaluate the activity of antimicrobial peptide PY on healthy human erythrocytes. D-galactosamine combined with LPS- induced mouse model of acute liver failure was constructed. The antimicrobial peptide PY effect on survival rate of mouse model was observed. HE staining was used to observe the pathological changes of liver tissue. Immunohistochemistry and Western blotting were used to detect the expression of apoptosis-associated protein caspase-3. Intra-group comparisons were performed using t-test and analysis of variance. χ (2) test was used for the comparison of rates. Results: An in vitro experiment showed that the endotoxin neutralization rate was higher at very low dose (0.01 μmol/L), and exceeded 70% at medium-dose (10-40 μmol/L), and the difference between groups with different concentration was statistically significant ( F = 569.22, P < 0.05). Medium-dose antimicrobial peptide PY had strong endotoxin neutralizing effect, low cytotoxicity and hemolytic activity. Moreover, in vivo experiments showed that the degree of liver injury and survival rate of mouse model was significantly improved with the medium-dose of antimicrobial peptide PY. Immunohistochemistry results showed that the expression of caspase-3 in the liver tissue was significantly depleted in the medium-dose group than that of the liver failure group, and the results were consistent with protein immunoblotting testing. Conclusion: Antimicrobial peptide PY possesses a strong ability to neutralize endotoxin and few toxic side effects. A specific dose of antimicrobial peptide PY can attenuate hepatocyte apoptosis and significantly improve the survival rate of animal model, and thus provides a new idea for the liver failure treatment.

Objective: To establish a model for predicting the short-term prognosis of patients with HBV-related acute-onchronic liver failure (HBV-ACLF) based on red blood cell distribution width (RDW) and the model for end-stage liver disease (MELD) scores., Methods: A total of 245 patients with HBV-ACLF were retrospectively analyzed for their clinical data and results of routine hematological tests, liver function, renal function, coagulation test, HBV-DNA, and other indicators at admission. Univariate analysis and binary logistic regression analysis were used to test the short-term risk factors for death of the patients, and the MELD-RDW model was established. The accuracy of each index and the established model was verified using the ROC curve., Results: The surviving patients with HBV-ACLF had significantly decreased RDW (14.97 ± 1.38) and MELD score (23.54±4.35) compared with those in the patients dead within 90 days (17.05±2.92 and 28.95±5.99, respectively). Multivariate analysis indicated that RDW was a significant independent prognostic factor for mortality in patients with HBVACLF (OR=1.840, 95%CI: 1.47902.289, P &lt; 0.005). The risk assessment model was [logisticMELD-RDW]=-9.375+0.582×RDW- 0.091×ALB-0.05×PTA+0.186×MELD. The area under the ROC curve of MELD score combined with RDW was 0.878, which was higher than RDW (0.724) and MELD score (0.780) alone., Conclusions: RDW is an independent prognostic indicator for mortality in patients with HBV-ACLF. Compared with MELD score, the risk assessment model based on MELD and RDW has a greater value in predicting the short-term prognosis of patients with HBV-ACLF.

Krüppel-like factors (KLFs) regulate diverse physiological processes such as the differentiation and development of red blood cells. However, it remains unclear whether KLFs exhibit synergistic regulatory effects. Transcriptomic data from our previous study showed that KLF1 and KLF9 expression was significantly higher in differentiated red blood cells than in hematopoietic stem cells. In the present study, we manipulated KLF1 and KLP9 gene expression by overexpressing or knocking down KLF1 and KLF9 in K562 cells and revealed a positive correlation between the expression of KLF1 and KLF9; their co-expression can significantly promote erythroid differentiation and specifically enhance β-globin gene expression. Further, we analyzed the transcriptome data of K562 cells with altered KLF1/KLF9 levels and found that KLF1 and KLF9 synergistically regulated erythroid differentiation through the PI3K-Akt and FoxO signaling pathways. KLF1 and KLF9 may exert this synergistic effect through FOS, TF, and IL8 in K562 cells. We have provided evidence that KLF1 and KLF9 play a synergistic role in regulating erythroid differentiation.

Objective: To study the clinical value of red blood cell distribution width (RDW) in the early prediction of acute kidney injury (AKI) in children with sepsis., Methods: A total of 126 children with sepsis were divided into an AKI group (n=66) and a non-AKI group (n=60) according to the presence or absence of AKI. These patients were also classified into high-RDW and low-RDW groups according to the mean RDW. The groups were compared in terms of age, male-to-female ratio, body mass index (BMI), Acute Physiology and Chronic Health Evaluation II (APACHE II) score, Sequential Organ Failure Assessment (SOFA) score, serum blood urea nitrogen (BUN), creatinine (Cr), uric acid (UA), serum C-reactive protein (CRP), and routine blood test results. Independent factors associated with RDW were analyzed by multiple linear regression., Results: Age, male-to-female ratio, BMI, CRP, SOFA score, and APACHE II score did not differ significantly between the AKI and non-AKI groups (P>0.05), but the AKI group had significantly higher BUN, Cr, UA, and RDW levels than the non-AKI group (P<0.05). Age, male-to-female ratio, and BMI did not differ significantly between the high-RDW and low-RDW groups (P>0.05), but the high-RDW group had significantly higher BUN, Cr, UA, CRP, SOFA score, APACHE II score, Hb, and mean corpuscular volume (MCV) than the low-RDW group (P<0.05). The multiple linear regression analysis showed that age, sex, APACHE II score, Cr, Hb, and MCV were independent factors associated with RDW., Conclusions: RDW has a certain clinical value in the early prediction of AKI in children with sepsis.

At present, the experimental technique to produce human red blood cells in vitro is complicated, and in order to optimize the induction steps, human pluripotent stem cells were differentiated into red blood cells through two induction steps. First, human pluripotent stem cells (including Rh negative A type umbilical cord mesenchymal stem cells (hUCMSCRh-A) and human iPS cells (hiPS)) were differentiated into CD31+ and CD34+ cells in BVF medium. PCR and flow cytometry were used to exam the expression of CD31 and CD34. We found that hUCMSCRh-A derived CD31+ and CD34+ cells were 5.3% and 22.7%, respectively; hiPS derived CD31+ and CD34+ cells were 31.2% and 8.2%, respectively. For the second induction step, the obtained CD31+ and CD34+ cells were differentiated into mature erythrocytes for 36 days under the addition of various growth factors. Through Giemsa staining, we found that the obtained mature erythrocytes were similar in morphology and size to normal human erythrocytes, and some obtained erythrocytes were enucleated. Globin expression was detected by real time RT-PCR, and the expression of β-globin was more than 20%. The obtained erythrocytes are collected into the centrifuge tube, and then erythrocytes were naturally settled and showed the red color. Our findings provide a novel and effective method for the quantity generation of human red blood cells in vitro.

Objective: To understand the changes in body weight, spleen weight and complete blood cells in BALB/c mice infected with Babesia microti ., Methods: For the infection group, six weeks old BALB/c mice were injected intraperitoneally with a dose of 100 μL of B. microti infected blood (20% RBC infection rate, each mouse). For the determination of the progression of B. microti infection up to 28 days of the infection, the microscopic visualization of thin blood smears of tail blood stained with Giemsa staining was performed in the infection group. The experiment was carried out at different intervals on days 0, 7, 14, 21, and 28 after the infection, respectively. The mice were sacrificed, and spleens were collected and weighed, and the body weight of the mice was also determined. The blood cells of the mice were analyzed by using Mindray BC-5300 Vet animal automatic hematology analyzer., Results: On the first day after the infection, B. microti was visualized in RBC of the infection group. The significantly highest infection rate (55%) appeared on the seventh day of the infection, and then steadily decreased; the mice attained the latent infection phase on the 28th day post-infection, when the parasite could not be visualized in the peripheral blood. The mice in the infected group acquired a significantly lowest body weight on the 7th day of the infection, and then gradually returned to normal. The weight of the spleen was the significantly highest on the 14th day of the infection, and then consistently decreased. On the 28th day of infection, the spleen weight was still higher than that of the control group. There were no significant changes in the number of white blood cells (WBC), lymphocytes, and eosinophils in the infected mice; and altered levels were all within the normal mouse reference range. The number of red blood cells, hemoglobin, and platelet count in the infected mice were decreased to the lowest level when the B. microti infection rate achieved to the highest, and then gradually returned to the normal levels., Conclusions: B. microti infection can cause body weight loss, splenic weight gain, and reduction in the number of erythrocytes and platelets in whole blood of the mice. Besides, the whole blood cell analyzer has a diagnostic significance in the identification of babesiosis.

Objective: To evaluate the efficacy of high (≥1:2) and low (<1:2) plasma: red blood cell (RBC) ratio resuscitation in patients with severe trauma requiring massive blood transfusion., Methods: The databases including the Cochrane Library, Pubmed, Web of Science, and EMBASE were systemically searched for relevant studies published between January, 2009 and April, 2016. The selection of studies, assessment of methodological quality and data extraction were performed by two researchers independently according to the inclusion and exclusion criteria. The main endpoint was 24-h mortality, 30-day mortality and 24-h survival rate., Results: Five observational studies reporting outcomes of 1024 patients were included in this meta-analysis. Four studies documented civilian cases and one study had a military setting. No significant differences were found in the Injury Severity Score (ISS) between patient groups receiving high and low plasma: RBC ratio resuscitation. Compared with the low-ratio group, the patients with high-ratio resuscitation showed a significant reduction in the 24-h mortality rate (OR=0.35, 95%CI [0.25, 0.48], P<0.000 01) and the 30-day mortality rate (OR=0.55, 95%CI [0.41, 0.75], P=0.0001). An increased survival rate was observed in patients receiving high plasma: RBC ratio resuscitation within the initial 24 h following the trauma (HR=2.34, 95%CI [1.46, 3.73], P=0.00001)., Conclusion: Raising the plasma: RBC ratio to 0.5 or higher may decrease the mortality rate of the patients with severe trauma who need massive blood transfusion.

Objective: To explore the size changes of erythrocytes infected with Plasmodium malariae , so as to improve the basic-level experimenters' microscopy capabilities for P. malariae identification in thin blood smears., Methods: The microscopic features of erythrocytes infected with P. malariae in thin peripheral blood smears were observed, and a microscope image processing software was used to measure and analyze the diameter changes of the erythrocytes infected with P. malariae., Results: The diameter of erythrocytes infected with P. malariae decreased significantly compared with that of the normal erythrocytes. The three parameters in this study: the diameter of erythrocytes, the value of diameter variation, and the ratio of diameter variation varied at different developmental stages of P. malariae , and the differences were statistically significant (all P < 0.01). The variances of the three parameters grouped by different cases or different developmental stages in different cases were analyzed, all showing statistically significant differences (all P < 0.01)., Conclusions: All the developmental stages of P. malariae will cause the decrease of the diameters of infected erythrocytes in peripheral blood smears, but the influence on the diameter of erythrocytes, value of diameter variation, and ratio of diameter variation varies at different developmental stages in different cases.

Objective: To study the association of red blood cell distribution width (RDW) and lipoprotein-associated phospholipase A2 (LP-PLA2) with the degree of coronary artery stenosis in patients with coronary artery disease (CAD) and the value of RDW combined with LP-PLA2 detection in accurate evaluation of coronary artery stenosis., Methods: A total of 224 patients including 119 non-CAD cases and 105 CAD cases admitted in our hospital between June, 2013 and June, 2014 were enrolled in this study. The patients' baseline clinical data were collected and venous blood samples were obtained for detecting WBC, RDW-CV and LP-PLA2. The Gensini score of the CAD patients was calculated based on coronary angiographic findings., Results: Compared with the non-CAD patients, CAD patients had significantly higher RDW-CV (P=0.009) and LP-PLA2 (P=0.004) levels. The CAD patients with high Gensini scores had also significantly higher RDW-CV (P=0.001) and LP-PLA2 (P<0.001) levels than those with low scores; RDW-CV and LP-PLA2 were significantly correlated with the Gensini score, and the area under curve of their combined detection was 0.931., Conclusion: Combination of RDW and LP-PLA2 can improve the diagnostic accuracy of the degree of coronary artery stenosis in patients with CAD.

Objective: To analyze the clinical, laboratory characteristics and PIG-A gene mutations in patients of myelodysplastic syndromes (MDS) with PNH clones., Methods: 218 MDS patients diagnosed from August 2013 to August 2015 were analyzed. The PIG-A gene mutations were tested in 13 cases of MDS with PNH clones, 17 cases of AA-PNH and 14 cases of PNH selected contemporaneously by PCR and direct sequencing., Results: 13 (5.96%) MDS patients were detected with PNH clones (13/218 cases). 9 patients were treated with cyclosporin A (CsA). Patients showed hematological improvement (HI). There were significant differences between MDS-PNH and PNH patients in terms of granulocyte clone size, red cell clone size and LDH levels [19.2% (1.0%-97.7%) vs 60.2% (3.1%-98.0%), P=0.007; 4.3% (0-67.2%) vs 27.9% (2.5%-83.6%), P=0.026; 246 (89-2014) U/L vs 1137 (195-2239) U/L, P=0.049], while the differences were not statistically significant in patients between MDS-PNH and AA-PNH patients [19.2% (1.0%-97.7%) vs 23.2% (1.5%-96.0%), P=0.843; 4.3% (0-67.2%) vs 14.4% (1.1%-62.8%), P=0.079; 246 (89-2014) U/L vs 406 (192-1148) U/L, P=0.107]. PIG-A gene mutations were detected in 7 MDS-PNH patients, of them, six were missense mutations, one were frameshift mutation and four cases with the same mutation of c.356G>A (R119Q). The PIG-A gene mutations were also detected in 9/11 AA-PNH patients and 11/14 PNH patients, both of them had the mutation of c.356G>A (R119Q). The PIG-A gene mutations of MDS-PNH, AA-PNH, PNH patients were all small mutations, the majority of those (59%) were missense mutation and mainly located in exon 2., Conclusion: MDS patients with PNH clones had better response to CsA, smaller PNH clone size. The PIG-A gene mutations of MDS-PNH patients mainly located in exon 2, which could be a mutational hotspot of these patients.

Objective: To establish a new method for detection of red blood cell osmotic fragility by using flow cytometry., Methods: The hypotension salt solution of different concentrations (0.70 ml normal saline+0.3 ml deionized water, 0.60 ml normal saline+0.40 ml deionized water and 0.55 ml normal saline+0.45 ml deionized water) were prepared with normal saline and deionized water, in which the red blood cells were suspended, and the residual red blood cells were detected by flow cytometer., Results: There was no significant difference in percentage of residual red blood cells between different time points detected by flow cytometer in 3 different hypotonic salt solutions. The percentage of residual red blood cells in B+C+D+E+F+G detected time region was different among 3 NaCl dilution groups. The percentage of residual red blood cells in normal control was lower than that in hemoglobinopathy group. The percentage of residual red blood cells in hereditary spherocytosis (HS) group was obviously lower than that in hemoglobinopathy and normal control groups. The comparison of 3 different dilution concentrations found that the second concentration (0.60 ml normal saline+0.40 ml deionized water) is more suitable to screen HS by FC500 flow cytometer., Conclusion: The detection of red cell osmotic fragility by using flow cytometry is a simple, rapid, objective and economic way that can be an effective screening method for diagnose the HS.

The implantable axial blood pump, driven by external electromagnet, is studied recently. It oscillats when it is running because of the elastic implanted environment and driving force disequilibrium, etc. In this paper, a model of single erythrocyte in vibrated flow field was built to simulate the deformation and force of the erythrocyte. By using the mechanical injury principle of blood in blood pump, we studied the injury of a single erythrocyte resulted from oscillating boundary flow field. The research results indicated that the shape of the erythrocyte, force and velocity field nearby, which are affected by oscillating boundary flow field, all cause injury to the erythrocyte. All the researches shown in the present paper are expected to provide theoretical foundation for lightening hemolysis by the blood pump.

To study the hemolytic effect of polyphyllin II (PP II) mediated by anion channel protein and glucose transporter 1 (GLUT1), in order to initially reveal its hemolytic mechanism in vitro. In the experiment, the spectrophotometric method was adopted to detect the hemolysis of PP II in vitro and the effect of anion channel-related solution and blocker, glucose channel-related inhibitor and multi-target drugs dehydroepiandrosterone (DHEA) and diazepam on the hemolysis of PP II. The scanning electron microscope and transmission electron microscope were used to observe the effect of PP II on erythrocyte (RBC) morphology. The results showed that PP II -processed blood cells were severely deformed into spherocytes, acanthocyturia and vesicae. According to the results of the PP II hemolysis experiment in vitro, the anion hypertonic solution LiCl, NaHCO3, Na2SO4 and PBS significantly inhibited the hemolysis induced by PP II (P < 0.05), while blockers NPPB and DIDS remarkably promoted it (P < 0.01). Hyperosmotic sodium chloride, fructose and glucose at specific concentrations notably antagonized the hemolysis induced by PP II (P < 0.05). The glucose channel inhibitor Cytochalasin B and verapamil remarkably antagonized the hemolysis induced by PP II (P < 0.01). The hemolysis induced by PP II could also be antagonized by 1 gmol x L(1) diazepam and 100 μmol x L(-1) DHEA pretreated for 1 min (P < 0.01). In conclusion, the hemolytic mechanism of PP II in vitro may be related to the increase in intracellular osmotic pressure and rupture of erythrocytes by changing the anion channel transport activity, with GLUT1 as the major competitive interaction site.

Objective: To investigate the effects of acute hypervolemic hemodilution (AHH) and intraoperative blood salvage (IOBS) on the morphology and biomechanics of erythrocytes using atomic force microscopy (AFM)., Methods: Blood samples were collected from 20 patients undergoing orthopedic surgery before operation (T1), immediately after AHH (T2), and after blood salvage (T3). AFM at nanometer resolution was used to examine the morphology and biomechanics of the collected erythrocytes., Results: The blood samples collected at T1 showed a significantly lower ratio of heteromorphous erythrocytes than those collected at T2 and T3 [(2.6∓1.3)% vs (19.3∓1.6)% and (17.6∓1.4)%, P<0.05]. AFM revealed significant differences in the morphology and biomechanics of the single erythrocyte in salvaged blood and blood after AHH compared with those of venous blood., Conclusion: AHH and IOBS may cause significant changes in the morphology and biomechanics of erythrocytes in the salvaged blood.

Most protocols for in vitro producing red blood cells (RBC) use the CD34(+) cells or embryonic stem cells from cord blood, bone marrow or peripheral blood as the start materials. This study was purposed to produce the mature RBC in vitro by using peripheral blood mononuclear cells as start material. The peripheral blood mononuclear cells (PBMNC) were isolated from buffy coat after blood leukapheresis, the mature red blood cells (RBC) were prepared by a 4-step culture protocol. The results showed that after culture by inducing with the different sets of cytokines and supporting by mouse MS-5 cell line, the expansion of PBMNC reached about 1000 folds at the end of the culture. About 90% of cultured RBC were enucleated mature cells which had the comparable morphological characteristics with normal RBC. Colony-forming assays showed that this culture system could stimulate the proliferation of progenitors in PBMNC and differentiate into erythroid cells. The structure and function analysis indicated that the mean cell volume of in vitro cultured RBC was 118 ± 4 fl, which was slight larger than that of normal RBC (80-100 fl); the mean cell hemoglobin was 36 ± 1.2 pg, which was slight higher than that of normal RBC (27-31 pg); the maximal deformation index was 0.46, which approachs level of normal RBC; the glucose-6-phosphate dehydrogenase and pyrurvate kinase levels was consistant with young RBC. It is concluded that PBMNC are feasble, convenient and low-cost source for producing cultured RBC and this culture system is suitable to generate the RBC from PBMNC.

Objective: To analyze specific expression of blood group genes using nucleated erythroid cells cultured from un-mobilized peripheral stem cells in vitro., Methods: Hematopoietic stem cells(HSC) bearing the CD34 antigen were isolated from peripheral blood by centrifugation and magnetic beads sorting, followed by suspension culture in vitro. Cells were collected from medium on various stages and analyzed by immunofluorescence. The RNA transcription of RH and ABO blood group genes was analyzed using culture cells on day 12., Results: A total of(3.19±0.13) ×10 (4) CD34+cells were isolated from about 50 mL peripheral blood with a recovery rate of 67.3%±2.7%. The cells amount in erythroid-lineage culture system on day 9 reached a plateau of a 237.1±15.5-fold amplification of the initial cell input. The stem cell-specific CD34 antigen was dropped off, while the erythroid-specific CD235a and CD240D antigens were increased in culture period. RHD/CE and ABO genes can be amplified using RNA extracted from culture cells on day 12, and genotypes of Rh and ABO systems by DNA sequencing were consistent with their serologic phenotypes., Conclusion: A method was established to analyze the gene expression of erythroid blood group derived from un-mobilized peripheral stem cells cultured in vitro. It can be used to study the expression of various erythroid-specific genes.

Erythrocytes lack nuclei and mitochondria, critical elements in the machinery of nucleated cell apoptosis. However, most recently, it became obvious that erythrocytes may undergo programmed aging, as well as suicidal death. The term eryptosis has been coined to describe the suicidal erythrocyte death. Eryptosis is triggered mainly by increased cytosolic Ca(2+) activity, in turn, Ca(2+) activates Ca(2+)-sensitive K(+) channels, scramblase, calpain and other proteases, respectively. A series of molecular events of erythrocyte programmed death induced. The cascade reaction of related molecules and finally lead to cell clearance. There is evidence suggesting that erythrocytes aging and death process are regulated tightly and there are many molecular participants and signaling pathways involved in aging and death process of erythrocytes. Erythrocytes have already been used as a model for aging study, and the knowledge about mechanisms involved in eryptosis may provide an important clue to understand the mechanisms involved in suicidal death of nucleated cells. In this review the factors influencing programmed death of erythrocytes, the role of Ca(2+) and ceramide in programmed death of erythrocytes, the role of blebbing in process of erythrocyte aging, the antigens of erythrocyte aging and so on are summarized.

In the present work, a cell environment which mimicked the real body environment according to the concentration radio between serum albumin and hemoglobin was built, and the cell morphology, the membrane deformation capacity, and the structure of intracellular hemoglobin of single human living erythrocyte under the effect of pH and serum albumin were studied. It was found that at different suspension pH, the magnitude of variations in cell shape and membrane deformation capacity changes with the structural changes of the intracellular hemoglobin. At pH 4. 14, 4. 76 and 10. 18, the loss of helical structure for hemoglobin, exposing of the hydrophobic amino acid in the globin chains, and changing of the combination of heme and globin, would completely destroy the stability of hemoglobin's structure, which seriously changes RBC's morphology and membrane deformation capacity. While at pH 6. 51 and 7. 80, the Raman spectra of erythrocytes are found to have no such changes, indicating that the structure of intracellular hemoglobin was not varied, thus the cell morphology and membrane deformation capacity are quite close to the normal values. At pH 5. 49 and 8. 76, RBC's morphology and membrane deformation capacity have different degrees of variation, but the structure of intracellular hemoglobin has not changed, suggesting that the cell morphology and membrane deformation capacity may be reversible. The results suggest that in the suspension solution containing serum albumin, erythrocytes have better ability to regulate and control the variation of the extracellular pH. In summary, upon building an environment which contains the same concentration radio of serum albumin to hemoglobin in the blood, this work performed systematic studies on the effect of pH on human erythrocytes. It can not only help to solve the problems about the mechanism of the structural and functional changes of erythrocytes induced by environmental pH, but also elucidates the possible variation of the erythrocyte under different pathologic states. It is therefore significant for the prevention and treatment of some diseases.

Objective: To investigate the effects of cisplatin plus hyperthermia on erythrocytes and killing human hepatocarcinoma (HepG2), gastro carcinoma (SGC7901) and colonic carcinoma (SW620) cells in the intra-operative blood salvage from cancer surgery in vitro., Methods: HepG2, SGC7901 or SW620 cells were mixed into the aliquot of erythrocyte concentrated from each intra-operative blood salvage of 30 patients subjected to gastrointestinal cancer surgery. The mixture cells were divided into the following groups (n = 30): A group (37 °C); B group (42 °C); C, D, E groups (50, 100, or 200 µg/ml DDP); F, G, H, I groups (42 °C, 25, 50, 100, or 200 µg/ml DDP). After treating for 60 min, tumor cells and erythrocytes were separated by density gradient centrifugation. The Na(+)-K(+)-ATPase activity, cell count, osmotic fragility, and blood gas variables were determined in erythrocytes. Cell viability and colony formation were determined in tumor cells., Results: Compared with A [(0.30 ± 0.08) µmol Pi/10(7)/h], the Na(+)-K(+)-ATPase activity was significantly decreased in E, H and I groups [(0.24 ± 0.07), (0.25 ± 0.06) and (0.24 ± 0.07) µmol Pi/10(7)/h] (P < 0.05). Extra-erythrocytic K(+) in E, H and I groups [(2.16 ± 0.37), (2.16 ± 0.38) and (2.56 ± 0.50) mmol/L] were significantly increased compared with A group [(1.53 ± 0.43) mmol/L] (P < 0.05). Compared with A group, osmotic fragility in E, H and I groups was significantly increased (P < 0.05). Among B, C, D, E, F, G groups, only in G group colony formations of HepG2, SGC7901, and SW620 (0% ± 0%, 0% ± 0% and 0.01% ± 0.01%) at 14 d were completely inhibited (P < 0.01) compared with A group (78.54% ± 7.83%, 72.28% ± 6.58% and 66.69% ± 6.69%)., Conclusion: Pretreatment with cisplatin (50 µg/ml) plus hyperthermia (42 °C) for 60 min in vitro might be an effective strategy to clear tumor cells contamination but preserve erythrocytes, which is worthy to be optimized and used in the intra-operative blood salvage in cancer surgery.

Red blood cell transfusion is an effective method to treat acute hemorrhage and severe anemia. However, blood source from donors is very limited, and transfusion-transmitted diseases occurred frequently, thus threatening human health. Therefore, the safe, abundant and functional blood source is needed. Generation of blood cells from human pluripotent stem cells(hPSC) will offer alternative approach. Lots of studies have been focused on erythroid cell differentiation in vitro, including how to enhance efficiency and improve their function. In this review, the research advances on differentiation methods and the regulatory mechanism are summarized. In addition, the progress in PSC differentiation into erythrocytes and the problems to be solved are discussed briefly.

This study was purposed to investigate the conditions for improving human-mouse xenograft and the erythroid differentiation of human hematopoietic stem cells (HSC) in the xenotransplant model. The engraftments of different mouse strains (NOD/SCID or NOD/SCID/IL2rγ(null)), schemes of irradiation (single-time or 2-times radiation; Co(60)γ-ray or X-ray) and strategies of CB CD34(+) cells ex vivo culture time and lentivirus infection were compared. The results showed that at 4 weeks after transplantation, the ratio of hCD45 positive cells in bone marrow of NOD/SCID/IL2rγ(null) mice increased to (51.4 ± 13.9)%, and erythroid precursor could be detected. All of the mice receiving X-ray irradiation for 2 times (a dose of 1 Gy, then the second of 1.5 Gy, with an interval of 15 min) survived. Fresh isolated CB CD34(+) cells were cultured and infected with lentivirus for 72 h and then transplanted into receptor mouse. After 4 weeks, higher engraftment [hCD45 (51.4 ± 13.9)%] and better erythroid development [hCD71(+) GPA(+) (5.98 ± 3.46)%] were observed. It is concluded that NOD/SCID/IL2rγ(null) mice receiving X-ray irradiation for 2 times and were injected with fresh isolated CB CD34(+) cells cultured and infected with lentivirus ex vivo within 72 h show a better xenograft and erythroid development.

Blood deficiency model of mice was copied by subcutaneous injection with 200, 100 and 100 mg x kg(-1) (0.01 mL x g(-1)) acetyl phenylhydrazine (APH) at the frist, fourth, and seventh days. Mice in each group were perfused with different extracted parts of Angelica sinensis (drug dosage was 2.4 g x kg(-1)) at the tenth day, once a day for 10 days. Then compare the influence of different extracted parts of Angelica sinensis to RBC, Hb, PLT and thymus, spleen and weight changes of blood deficiency mice. The peak areas of each common peak from HPLC fingerprint were associated with the date of replenishing blood pharmacodynamics efficacy by using gray relation statistic, which was used to research the chromatogram-pharmacodynamics relationship. The results showed that the part of DSC has the better effect in replenishing blood. The contribution degree of the DSC to replenishing blood of each component were determined by correlation size, and ferulic acid made the largest contribution, but contribution of other components should not be ignored. In this paper, we research the relationship of the HPLC fingerprint and spectrum activity relationship, determine the material basis of the DSC for replenishing blood, and provide effective way to represent the spectral correlation effect.

This study was purposed to investigate the relationship between HLA-A, B allele polymorphisms and red blood cell parameters of patients with --(SEA/αα) subtype of α(0)-thalassemia in Han ethnic population of Wuzhou city. The HLA genetic polymorphisms were determined by polymerase chain reaction-sequence-based typing (PCR-SBT) in 57 patients with --(SEA/αα) subtype of α(0)-thalassemia of Han ethnic population in Wuzhou city, Guangxi province, China. Mean corpuscular volume (MCV), hemoglobin (Hb), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) were detected by automatic blood analyzer system. HbA2 were detected by electrophoretic method. The statistical analysis was performed by ordinal polytomous logistic regression. The results showed that Hb and HbA2 levels were significantly lower in patients positive for HLA-A*33:03, B*15:01 or B*55:02, and were significantly higher in patients positive for B*15:02 (P < 0.05). It is concluded that several HLA alleles may be associated with Hb level of --(SEA/αα) subtype of α(0)-thalassemia of Han ethnic population in Wuzhou city. This result has the value for understanding phenotype-genotype relationships in thalassemia.

Objective: To prepare stable chicken red blood cells for the calibration of flow cytometry., Methods: The traditional isolation method of chicken red blood cells was modified by incorporating gelatin technique, Ca2+-free HBSS treatment and low-speed centrifugation. The effect of fluorescence staining of the cells was improved by the addition of TritonX-100 to enhance the membrane permeability and Rnase enzymes to disintegrate RNA tiles. The modified method was compared with the traditional method for viability of the freshly isolated cells and the DNA content coefficient of variation (CV) of the fixed cells., Results: Chicken red blood cells obtained by the modified method showed a significantly higher viability than those obtained by the traditional method [(98.5∓3.5)% vs (93.5∓2.7)%, P<0.05]. After glutaraldehyde fixation, the isolated cells with the modified method were stable during the 90-day preservation with a significantly lower CV than the cells obtained by the traditional method [(6.0∓0.3)% to 6.2∓0.4% vs (8.6∓0.5)% to (13.1∓1.4)%, P<0.01]., Conclusion: The chicken red blood cells isolated using the modified method can be applicable for calibration of flow cytometry.

Objective: To investigate the effects of living high-training low (HiLo) on innate immunity in blood of elite swimmers., Methods: Six female swimmers undertook HiLo for two weeks, erythrocyte adhesion function and counts of leukocyte were tested in different time of training period., Results: Red blood cell C3b receptor ring rate (RBC-C3bRR) decreased and red blood cell immune complex matter ring rate (RBC-ICR) increased significantly (P < 0.05), the two markers returned to base line 1 week after training. Counts of leukocyte and granulocyte decreased significantly (P < 0.05), and they recovered 1 week after training; Counts of lymphocyte and monocyte decreased without significance during training and did not recovered after training., Conclusion: Immunity of erythrocyte and granulocyte decreased quickly, but lymphocyte and monocyte recovered slowly, swimmers were adaptive to the training.

This paper was to explore the effect of blood oxygen saturation (SO2) on oxidative damages of erythrocytes under the condition of oxidative stress. Keeping SO2 of cultured erythrocytes in vitro at the states of 0.3, 0.5, 0.7, 0.9 and 0.98, respectively, we induced oxidative stress by tert-buthylhydroperoxide (BHP, 0.15 mmol/L of final concentration). After incubation, antioxidant capacity was assessed by measuring content of reduced glutathin hormone (GSH) in erythrocytes. Methemoglobin (MetHb) content, lipid peroxidation (thiobarbituric acid-reactive substances, TBARS) and denatured globin-chains on the plasma membrane were measured to assess the extent of oxidative damages. The results showed that in the presence of BHP, GSH contents increased from 0.3 to 0.98 groups; MetHb, TBARS and globin-chains levels all dropped with the rise of SO2. In conclusion, antioxidant capacity and oxidative damages of erythrocytes are closely related to SO2, declined SO2 could promote oxidative damages of erythrocytes.

Objective: To study the effects of hermap gene on kinases in erythroid signal transduction pathway and investigate the mechanism of hermap on erythroid differentiation., Methods: The K562 cells expressing hermap and hermap-siRNA respectively were established for up- and down-regulating the expression of hermap gene. These K562 cells were then induced by Ara-C to erythroid differentiation and analyzed at 0, 24, 48, 72 and 96 h, respectively, for cell morphology and biphenylamine staining positive cells, determination of CD235a, CD36, kinases p-STAT5, p-Akt, p-MAPK and p-c-JUN by FCM; and quantification of hermap gene and γ (Aγ,Gγ) globin gene by FQ-PCR., Results: With up-regulating hermap gene and inducing by Ara-C, K562 cells were changing to low ratio of nucleus to cytoplasm, cytoplasm colour from basophilic to pinkish or amethyst tinge, increase of number of biphenylamine positive cells and expression of CD235a, CD36, γ (Aγ,Gγ) globin gene, hermap gene and p-STAT5 from 0 to 96 h. At 0, 24, 48, 72 and 96 h of culture, the positive rates of p-STAT5 cells were detected of 0.46%, 4.54%, 20.01%, 23.65% and 33.08%, respectively. This results demonstrated that there was a positive correlation between expression of p-STAT5 and hermap gene expression (P < 0.05)., Conclusion: hermap gene can stimulate erythroid differentiation of Ara-C induced K562 cells mainly through JAK/STAT5 signal transduction pathway.

The purpose of this study was to investigate the distribution of 10 rare red blood groups in Chinese Nanjing population, so as to provide compatible rare blood to patients and to create a donor data bank. Jk (a-b-) (Kidd) phenotypes were detected by urea, while H-(H), GPA-(MNS), GPC-(Gerbich), i+ (Ii) and Lub-(Lutheran) phenotypes were detected by monoclonal, polyclonal antibodies with U type 96 well microplate technology. The screening of Jsb- and k-(Kell), Fya-(Duffy), Ok-(Ok), s-(MNS) and Dib-(Digeo) phenotypes were performed by polymerase chain reaction. The results showed that 2 Jk (a-b-) out of 40337 donation samples and 3 Fy (a-b+) out of 1782 donation samples were found, while no other rare blood phenotypes (H-, GPA-, GPC-, Lub-, Ok-, s-, Jsb-, k-, Dib- and i+) were detected. It is concluded that the frequencies of Jk (a-b-) and Fya(a-b+) are 0.0049% and 0.168% respectively. No more rare blood phenotype was found in this screening.

Objective: To investigate the different parameters of the lyophilization procedures that affect the recovery of the rehydrated red blood cells (RBCs)., Methods: Human RBCs loaded in tubes were cooled with 4 different modes and subjected to water bath at 25 degrees celsius;. The morphological changes of the RBCs were observed to assess the degree of vitrification, and the specimens were placed in the freeze-dryer with the temperature set up at 40, -50, -60, -70 and -80 degrees celsius;. The rates of temperature rise of the main and secondary drying in the lyophilization procedures were compared, and the water residue in the specimens was determined., Results: The protectant did not show ice crystal in the course of freezing and thawing. No significant difference was found in the recovery rate of the rehydrated RBCs freeze-dried at the minimum temperature of -70 degrees celsius; and -80 degrees celsius; (P > 0.05). The E procedure resulted in the maximum recovery of the RBCs (83.14% ± 9.55%) and Hb (85.33% ± 11.42%), showing significant differences from the other groups(P < 0.01 or 0.05). The recovery of the RBCs showed a positive correlation to the water residue in the samples., Conclusion: Fast cooling in liquid nitrogen and shelf precooling at -70 degrees celsius; with a moderate rate of temperature rise in lyophylization and a start dry temperature close to the shelf equilibrium temperature produce optimal freeze-drying result of human RBCs.

Objective: To study the changes of rat red blood cells on T, B lymphocytes proliferation in chronic unpredictable mild stress (CUMS)., Methods: Thirty Wistar rats were averagely divided into control group and the CUMS group. The relevant behavioral performance and red blood cells on lymphocytes proliferation Changes by MTT colorimetric were observed., Results: After fourteen days, the growth of weight and organic coefficient of brain decreased in model group (P < 0.05), with a significantly reduced consumption and preference of sucrose solutions, and increased pure water consumption as compared with control group (P < 0.05). Plasma corticosterone levels peaked at seven days but on a declining trend after fourteen days. The rate of red blood cells on T, B lymphocyte proliferation in depression group was lower than non-stress in 7, 14, 21 days (P < 0.05)., Conclusion: The repeated chronic mild stress stimulation can induce long-time changes in acts and activities and decrease the rate of red blood cells on T, B lymphocytes proliferation, which can provide a new experimental method to observe the red blood cell immune function under the stress.

Erythrocyte is a mature blood cell that contains hemoglobin to carry oxygen to the bodily tissues. Erythrocyte, which takes on a biconcave disc that has no nucleus, is flexible and changeful. Erythrocyte is so sensitive to the environment that the shape of cell goes crimpy, even acanthoid. A laser tweezers Raman spectroscopy (LTRS) setup was used to trap single erythrocyte from healthy donors and patients with thalassemia and to collect the Raman scattering of trapping cell. Normal shape, crimpy erythrocytes and acanthoid erythrocytes were tested, and the averaged spectra, and principal component analysis (PCA) which detailed the spectral difference and the change of hemoglobin, were used to evaluate the effects of different cell shape on the spectral distinguishing of erythrocyte. The results reveal that in normal physiological environment the change in cell shape does not effect the spectral distinguishing of abnormal erythrocyte.

Objective: To observe the effects of boron supplemented in water on the blood composition in rats., Methods: 192 Sprague-Dawley rats (28 +/- 2) d were randomly divided into six groups (n=32, 16 male rats and 16 female rats), and drank the distilled water supplemented with 0, 40, 80, 160, 320 and 640 mg/L boron for 60 days, respectively. At the 30th and the 60th day of experiment period, 16 rats (n=8, male=female) from each group were selected and made into narcosis with 10% Chloral Hydrate. Then the blood were obtained from the right atrium after dissection, the blood smear were made and stained with Wright's and Giemsa's stains at the same time. Then the red cells count, the white cells count, the reticulocyte counts, the hemoglobinometry and the differential leukocytes count were determined., Results: In comparison with control group, the reticulocyte percentages of all experiment groups exhibited different degree increases, the RBC numbers and Hb contents in rats consuming 40 and 80 mg/L of boron more significantly raised (P < 0.05), but the changes of the leucocyte percentages were more no significant. The numbers of RBC and WBC and the Hb contents in rats supplemented with from 160 to 640 mg/L of boron all decreased significantly or most significantly (P < 0.05 or P < 0.01), and the percentages of neutrophil, eosinophil, basophil and monocytic showed similar changes on the 60th day of experiment time., Conclusion: The distilled water supplemented with 40 mg/L of boron could promote the hemopoiesis and Hb synthesis in rats, and the drinking water supplemented with 80 mg/L of boron would have some promotion on the erythropoiesis and Hb synthesis; but when the boron supplementation from 160 to 640 mg/L, the higher doses boron could inhibit the hemopoiesis and Hb synthesis obviously, or even exhibit apparent toxicity action.

Aim: To investigate the relationship between changes of peripheral blood counts and tumor rejection induced by a low dose of melphalan in C57BL/6 mice., Methods: Mouse lymphoma EL4 cells were inoculated subcutaneously into wild type C57BL/6 mice. Twelve days later, 7.5 mg/kg melphalan were administered intraperitoneally and the same volume of Normal Saline as control. Tumor sizes were observed and recorded subsequently. Blood samples were obtained from orbital venous sinus on different days before and after melphalan treatment, and then complete blood counts were performed and the relationship between the alterations of blood counts and tumor shrinkage after melphalan treatment was analyzed., Results: Tumor sizes decreased and tumors disappeared after 7.5 mg/kg melphalan treatment; while tumors grow continuously in control mice. The number of WBC was increased a little (10.6 + or - 2.3) x 10(9)/L 6 h after melphalan treatment, but there was no significant difference with mice before melphalan injection (9.8 + or - 0.32) x 10(9)/L (P>0.05); The number of WBC decreased significantly at 4(th) day after melphalan treatment (P<0.01); Later it increased a little, but at 28(th); day after melphalan it still obviously lower than that of the normal (P<0.01). Hemoglobin (Hb) concentration decreased from (132 + or - 7) g/L before melphalan treatment to (110 + or - 14) g/L at 6 h after melphalan treatment (P<0.05). Later, the amount of Hb was decreasing and at 7th day it got to its lowest point (96 + or - 5) g/L. It increased gradually back to normal in 2 weeks after melphalan treatment. The platelet count increased to (1502 + or - 142) x 10(9)/L 6 h after melphalan treatment, significantly higher that that (914 + or - 322) x 10(9)/L before melphalan injection (P<0.01). It maintained at a high level for one week and it recovered back to normal level at 28(th) day after melphalan treatment., Conclusion: Tumor shrinkage after melphalan treatment was not related to the decreased number of WBC or RBC, but correlated with the increased number of platelet in 10 days after melphalan treatment.

Objective: To explore the influence of carbogen on lateral wall microvascular of cochlear after acute acoustic trauma., Method: Forty guinea pigs were divided into 4 groups: group A (noise damage), group B (carbogen inhalation), group C (noise damage + carbogen inhalation), and the control group without any treatment. The diameter of the column of RBCs (RBC column diameter, RBCCD), blood flow velocity (BFV) and blood flow states(BFS) in microvasculature were measured and described under microscope., Result: The microvascular in group A demonstrated a blood flow in contrary direction, granuliform flow, and granular slow flow. The erythrocytes aggregated in the microvascular of the cochlea. The RBCCD decreased 12.1% compared with the control group (P < 0.05). The blood flow in group B showed a laminar flow or laminar granular flow, and the RBCCD increased 20.7% compared with the control group. The blood condition in group C was the same as the control group-laminar granular blood flow; the blood flow with contrary direction was less than group A, and the RBCCD was 17.4% lager than that of group A., Conclusion: Carbogen can dilate the RBCCD and increase the BFV in stria vascular. So carbogen can alleviate the harm from noise.

The technique of confocal Raman scanning microscopy (point scanning, line scanning and 2D scanning) and bright field imaging of living erythrocytes was investigated as a function of different scanning conditions at the excitation wavelength of 514 nm. The biological effect of the 514 nm laser radiation on the erythrocytes was also evaluated, so that a set of proper scanning parameters for different scan modes can be determined to obtain strong enough Raman signal while without damage on the living cells by evaluating the change of Raman spectra and lighted field images of the cells. For the point scanning mode, the laser power at sample is the most important parameter to be adjusted, which normally should be less than 1.5 mW. For the line scanning mode, the laser power at sample and scanning step should be considered at first. Small scanning step means the energy of laser accumulated at a small region, which can easily damage to erythrocytes. Large scanning step can reduce the damage; however the spatial resolution decreases also. It is recommended that scanning step should be more than 0.5 microm and laser power at sample should be less than 0.7 mW. For the 2D scanning mode, besides the laser power at sample, scan step needs to be adjusted, and other scan parameters need to be adjusted properly for reducing the effect of laser on erythrocytes. Large pinhole and relative low temperature of sample are the remedies, which can reduce the effect of laser on erythrocytes. 1.0 microm scanning step, 0.7 mW laser power at sample, 500 microm pinhole and proper low temperature can get better 2D Raman image of erythrocytes. For all scanning modes, if the Raman signal is strong enough, the exposure time can be shortened properly, thus reducing the effect of laser on erythrocytes. The optimization of experiment process is also important for Raman test on living cells.

In order to elucidate the expression and molecular genetic background of ABO gene seven samples with ABO discrepancy further identified as bi-specific ABO gene were studied. All these samples were subjected to phenotyping by monoclonal and polyclonal antisera and were then genotyped by direct DNA sequencing and haplotype-sequencing at the exon 6 and 7 of ABO gene. As a result, six ABO dual-specific alleles were identified in Chinese population. An antigen expressed by these B (A) or Cis-AB individuals varied from very low level to the normal level, compared with common A blood group samples. In conclusion, molecular genetic backgrounds of two pairs out of four samples in all samples were the same, however, the ABO expression showed diverse.

This study was aimed to compare the difference of quality between buffy-coat-collected platelet concentrates (BC-PC) and single-donor plateletpheresis (SDP). 15 packs of BC-PC and 15 units SDP were stored at 20 degrees C - 24 degrees C with agitation. Platelet concentration, platelet volume, residual leukocyte and residual erythrocyte in two groups were examined after preparation for 1 hour. Mean platelet volume, pH value, hypotonic shock response (HSR), CD62p expression and CD62p re-expression of platelet were detected on 0, 1, 2, 3, 4, 5 days of platelet preservation. The results showed that the platelet yields, residual leukocyte and residual erythrocyte in two groups accorded with the national quality standard respectively, but residual leukocyte and residual erythrocyte in BC-PC group were higher than those in SDP group when platelet yields in two groups were equal (p < 0.01). Lactate concentration, CD62p expression of platelet increased with prolongation of preseved time, while pH value decreased gradually. Compared with SDP group, there were significant differences in CD62p expression, CD62p re-expression of platelet preserved for 0 - 5 days (p < 0.01), and in pH value of platelet preserved 2 - 5 days (p < 0.01). There was no changes in HSR of SDP group for 0 - 5 days, while HSR in BC-PC group decreased gradually. There were significant differences in HSR of platelet preserved for 1 - 5 days (p < 0.01). It is concluded that the platelet concentrates prepared by BC-PC are not equal to SDP in quality, the preparation technology of BC-PC should be optimized further in order to reduce residual leukocyte, residual erythrocyte and activated platelet yields, as well as improve the quality of BC-PC.

Objective: To study the erythrocyte immuno-regulatory effect of Patrinia scabra Bunge extracts extracted by macroporous adsorptive resins in tumor bearing mice., Methods: Patrinia scabra Bunge was extracted by macroporous adsorptive resins, and the amount of polysaccharides and saponins in the extract were determined. Mice bearing S180 tumor were treated with the extract and their survival prolongation rate, erythrocyte rosette formation rates of C3b receptor (ERR-CR), immune complex (ERR-IC) and tumor cell (ERR-TC), as well as the CD35 and CD44s were observed., Results: Polysaccharide content was 21.4%, saponin 41.8% in the extract. As compared with the model group, the survival rate was increased, the erythrocyte immune function was improved (showed increase of ERR-CR and ERR-TC, decrease of ERR-IC), and the amount of CD35 and CD44s in red blood cell membrane increased in mice after being treated with the extract (P < 0.05 or P < 0.01)., Conclusion: Extract of Patrinia scabra Bunge extracted by macroporous adsorptive resins can regulate the erythrocyte immune function to a certain extent.

This study analyzed the effect of Sargassum fusiforme polysaccharides (SFPS) on the complex mobility of erythrocytes in tumor-bearing organisms. A mouse tumor model was developed in which mice were intraperitoneally injected with high, medium, and low dosages of SFPS for 7 d. The erythrocytes were collected and prepared into suspensions, and the complex mobilities of cells were measured using high performance capillary electrophoresis (HPCE). Experimental conditions included the following: capillaries, 75 microm x 50 cm; buffer for electrophoresis, phosphate solution containing hydroxypropylmethyl cellulose (0.1 mol/L, pH 7.4); injection pressure, 3.448 kPa; injection time, 10 s; separation voltage, 20 kV; column temperature, 25 degrees C. The electroosmotic mobility was calculated to be 2.16 x 10(-4) cm2 x V(-1) s(-1). It was found that SFPS can decrease the migration time of erythrocytes in tumor-bearing organisms, which was 17.56 min for the control group, 13.96 min for the low dosage group, 12.90 min for the medium dosage group, 13.51 min for the high dosage group, and 11.51 min for the normal group, and increase its complex mobility, which was 1.06 x 10(-4) cm2 x V(-1) x s(-1) for the control group, 1.19 x 10(-4) cm2 x V(-1) x s(-1) for the low dosage group, 1.29 x 10(-4) cm2 x V(-1) x s(-1) for the medium dosage group, 1.23 x 10(-4) cm2 x V(-1) x s(-1) for the high dosage group, and 1.45 x 10(-4) cm2 x V(-1) x s(-1) for the normal group. Compared with the negative control group, all three dosage groups had significant differences (P < 0. 01) in the complex mobility of erythrocytes. It was found that SFPS can change the complex mobility of erythrocytes in tumor-bearing organisms with the tendency to the condition in normal organisms, which is possibly because SFPS can change the charges density on erythrocytes surface. It is believed that HPCE can be used as an auxiliary tool for determining the physiological state and functions of erythrocytes.

Objective: To discuss the effect of Yisui Shengxue granules on expression of alpha-hemoglobin stabilizing protein (AHSP) mRNA in different developmental stages mice., Method: The total RNAs were extracted from the bone marrow karyocyte of normal adult mice and the karyocyte of fetus liver and fetus spleen in pregnanted mice (pregnanted 21 days) and fetal mice (pregnanted 14 days). The expression level of AHSP mRNA in different developmental stages mice interfered with Yisui Shengxue granules was measured by real-time PCR., Result: The intervention of Yisui Shengxue granules could significantly up-regulated the expression levels of AHSP mRNA in normal adult mice., Conclusion: The result revealed that one of possible molecular mechanism of the effects caused by Yisui Shengxue granules is that it can promote the AHSP gene expression, reduce the free a-globin deposit, then prevent the poison to erythrocyte and decrease the haemolysis.

Laser tweezers Raman spectroscopy (LTRS) system is a combination of spectroscopy and laser tweezers; It is a new method of studying cells; It can trap single living cell and make Raman spectrum of single living cell. From the positions, intensities, and line widths of the Raman peaks in the spectra, we can get useful information about composition, structure and interactions of complexes inside the living cells. External agents may change cell's physiological state and this changed information can also be got from Raman spectra. This article is a study of Raman spectra of human red blood cell (RBC) affected by different intensity direct current (DC); from the result, distinct change of Raman spectra of RBC have been got. These changes characterize the changes of the internal information of the cells. This article give some academic reference of physical therapy using DC in the level of molecule.

Objective: To clone and express avian influenza A virus [A/Hong Kong/482/97(H5N1)] H5 subtype hemagglutinin in baculovirus-insect cell expression system and investigate the antigenicity and bioactivity of the recombinant protein., Methods: H5 gene of influenza A virus was amplified by PCR. The recombinant bacmid was obtained by cloning the gene to the donor plasmid of pFastBacHTB and transformed into DH10Bac competent cells. The recombinant baculovirus stock was prepared by transfecting the recombinant bacmid DNA into the insect cell line for protein expression after amplification. Immunofluorescene assay (IFA) and Western blotting were performed to identify the antigenicity of the recombinant protein, and hemagglutination assay was used to identify its bioactivity., Results: The recombinant his-H5 protein was expressed in the insect cells with a relative molecular mass of 64,000, which showed erythrocyte-agglutinating activities with the red blood cells of guinea pig. Western blotting and IFA demonstrated that the recombinant his-H5 could be recognized and bound by standard anti-H5 sera., Conclusion: The recombinant his-H5 with a post-translation modification is successfully obtained in insect cells, which may provide a potential source for further study of the antigen's biological function and for production of the subunit vaccine or monoclonal antibodies.

Agglutinin with hemagglutinating activity was found in hemolymph of Tegillarca granosa Linnaeus. By extraction, fraction with saturated ammonium sulfate precipitation, gel filtration on Sephadex G-100 and followed by affinity chromatography on Sepharose 4B, an agglutinin was purified from the hemolymph of T. granosa. The purified lectin contains 5.02% neutral saccharide. The molecular weights is about 123 KDa, which showed 2 bands on SDS-PAGE, and the relative molecular weights of these subunits were 15 KDa and 16 KDa, respectively. In its amino acid composition, Asp has the highest content, followed by Glu and His, but there is a lack of Met. Assays for erythrocyte agglutination and sugar inhibition were done in microtiter plates. The result indicared that TGL-hemolymph could agglutinate erythrocytes from the natural and enzyme-treated erythrocytes of human and animals, and the highest activity was found in the agglutination with rabbit erythrocytes. The agglutinic activity of TGL-hemolymph was inhibited by lactose and D-galactose. The hemagglutination activity of TGL-hemolymph depended on Ca2+, and it showed the highest hemagglutination activity in pH 7.0. Thermostability was not high. The agglutinating activities changed from 2(5) to 2(1) between 30-70 degrees C, and was completely destroyed beyond 80 degrees C.

Trehalose, as a nonreducing disaccharide, plays a role in protecting the cytoactivity when the cells is freezing, drying or lyophilization. It has been a biomembrane protectant applied to lyophilization of human blood cells (platelets and erythrocytes), and from which astonishing results have been obtained. Having powerful hydration, distinctive vitrification transform and crystal transform and unique resistance of high temperature and humidification, trehalose is thought of a preferred protectant in the study of cell preservation. In recent years, people concerned trehalose on its protective mechanism, experimental means of transit trehalose to mammal cells and the mechanism of loading in red blood cells. The above aspects were briefly summarized in this article.

In order to investigate the effect of antioxidants on human blood, vitamin C was selected and added into plastic blood storage bags with CPD, and stored at 25 degrees C. During 6 days of storage, some indexes as ATP, SOD, MDA, K(+) concentration and superoxide radicals were detected and were compared with control group, The results showed that ATP and SOD activity in whole blood with vitamin C during 6 days of storage was higher then that in control group (P < 0.05, P < 0.01), the MDA and plasma K(+) concentrations in stored whole blood with vitamin C during 6 days of storage were lower than that in control group (P < 0.05, P < 0.01), the superoxide radical concentrations in stored whole blood with vitamin C decreased lower than that in control group (30%). The conclusion was made that vitamin C increases activities of ATP and SOD, decreases concentrations of MDA, plasma K(+) and superoxide radicals during blood preservation.

Objective: To identify anti-hypoxia ingredients extracted from Portulaca oleracea and to find out the possible mechanism of its anti-hypoxia actions., Methods: Seventy mice were randomly divided into seven groups which were untreated (normal saline), ginsenosides-treated, polysaccharide-treated, acidic components-treated, basic components-treated, alkaloids-treated and flavones-treated groups, and the ingredients of polysaccharide, acidic components, basic components, alkaloids and flavones were extracted from Portulaca olerace. The mice in each group were fed with corresponding ingredients for one week respectively. Then the survival time of mice in hypoxic conditions was observed. Another 90 mice were divided into 3 groups: untreated (normal saline), ginsenosides-treated and flavones-treated groups. The mice in each of these 3 groups were divided into 3 subgroups according to 12-, 24- and 36-hour exposure to hypoxia (10% oxygen and 90% nitrogen), respectively. After exposure to hypoxia, the red blood cell count (RBC), hemoglobin (Hb) concentration and hematocrit (HCT) in mice were determined. The plasma erythropoietin (EPO) levels of mice were detected by enzyme-linked immunosorbent assay (ELISA) and the relative values of EPO mRNA in renal tissue and pallium of mice were determined by reverse transcriptase-polymerase chain reaction (RT-PCR)., Results: The survival time of mice in hypoxic conditions in flavones-treated group was significantly longer than that in the untreated group. The RBC, Hb concentration, HCT, plasma EPO level and the relative values of EPO mRNA in renal tissue and pallium of mice were significantly higher in the flavones-treated group than those in the untreated group., Conclusion: The anti-hypoxia ingredients extracted from Portulaca oleracea are flavones and the anti-hypoxia effects may be obtained by improving the expression level of EPO and accelerating the generations of erythrocyte and Hb.

Objective: To establish flow cytometry (FCM) methods and evaluate their application value for measuring the index for enhancing immune function of health food., Methods: In mice experiment model, the dosage groups were respectively oral fed with three test substances according to 5, 10, 30 times of the recommended dose for human body; both the negative and positive control groups were fed with equivalence purified water once a day. The positive control was fed with 25 mg/kg body weight levamisole for 3 days before finishing the administration, and the immune two percent of sheep erythrocytes were administrated at the last day. In rats experiment model, the test substance was given by mixing feed according to 25 and 50 times of the recommended dose for human body. At the end of the experiment, indices below were simultaneously detected. (1) The classical indices included: spleen lymphocyte transformation test by using ConA (MTT assay); spleen NK cell activity test (LDH assay); delayed-type hypersensitivity test by using sheep erythrocyte (foot palm thickening) method and phagocytosis activity tested by mice peritoneal macrophages. (2) FCM indices included: T and B lymphocytes quantitating in mice peripheral blood, activated antigen expression level in the surface of T lymphocytes and NK cells and phagocytosis activity for fluospheres in mice peritoneal macrophages., Results: (1) Compared with the negative control group, there were no significant differences in T and B lymphocytes proportion and the number of lymphocytes in mice peripheral blood after given 0.83, 1.67, 5.01 g/kg protein powder; (2) mice peripheral blood T lymphocyte sub-cluster CD(69)(+)/CD(3)(+) of 3.75, 7.50, 15.0 ml/kg bw Cen-Rong Cream groups were all significantly increased (P < 0.05), which were shown a good coherence with the classic test index; (3) mice peripheral blood NK cell sub-cluster CD(69)(+)/NKG2D(+) of 0.83, 1.67 g/kg protein powder groups were both significantly increased (P < 0.05), which was kept in good coherence with those of NK cell activity test (LDH assay); rats peripheral blood NK cell sub-cluster CD(161a+)/CD(25)(+) of 1.50 g/kg ganoderma lucidum and cordycepicmycelia group was significantly increased (P < 0.05); (4) the phagocytosis activity in mice peritoneal macrophages: there were no significant difference found between the controls and the dosage groups in the classic test. However, in the FCM test, the percentage of phagocytic cells of 0.15, 0.30, 0.90 g/kg ganoderma lucidum and cordycepicmycelia groups and the phagocytic index of 0.30, 0.90 g/kg were enhanced., Conclusion: It suggests that was shown in detecting and assessing enhancing immune function of health food the results tested by FCM were fairly consistent with those by using traditional methods, most of them would have higher sensitivity. It should be valuable to applying FCM in the measurement and assessment of enhancing immune function of health food and worth while to further study as to enlarging its application.