Effects of loaded buffer with epigallocatechin gallate on physiological functions of platelets / 中国实验血液学杂志

This study was aimed to explore the change of aggregation and activation of platelets loaded with epigallocatechin gallate (EGCG). The platelets were treated by loading buffer with different concentrations of EGCG (0, 2.5, 5, 7.5, 10, 12.5, 15, 20 and 30 mmol/L) and were divided into 2.5, 5, 10 mmol/L groups and control group. The physiological and biochemical functions of platelets were observed, including recovery rate, aggregation and activation of platelets. The platelet counts were determined by Counter Cell-DYN 1200. The aggregation activities were tested through turbidimetry, the platelet apoptosis was detected by flow cytometry. The results showed that the concentrations of EGCG loading in platelets of 2.5, 5 and 10 mmol/L groups were 0.4006 ± 0.12, 1.0527 ± 0.1503, 1.6902 ± 0.1112 mmol/L respectively. Along with the increasing of EGCG concentrations in loading-buffer, the EGCG absorbed by platelets increased too. When the concentration of EGCG in loading-buffer exceeded 15 mmol/L, the EGCG absorbed by platelets did not increase. The recovery rate in 2.5 mmol/L loading buffer group was 82.45 ± 0.360% which was lower than that in control group (90.33 ± 1.115%) (p < 0.05). As compared with control group, the recovery rate in 5 mmol/L loading buffer group (57.51 ± 2.468)% and 10 mmol/L loading buffer group (47.45 ± 2.030)% were even significantly lower (p < 0.01). When ADP was used as the inducer, the maximal aggregation rate (MAR) in control group was (63.6 ± 4.037)%, which was higher than that in other EGCG-loading groups (p < 0.01). And the aggregation activity of platelets negatively correlated with the concentration of EGCG in loading-buffer. When THR was used as the inducer, the MAR in control group was (89.3 ± 6.533)% and higher than that those in other groups (p < 0.05), especially in groups with loading-buffer higher than 10 mmol/L EGCG (70.1 ± 5.400%) (p < 0.01). In the experiment of cellular apoptosis, the early apoptosis easy appeared in platelets loaded with EGCG. It is concluded that the EGCG loading in platelets markedly influences the physiological and biochemical functions of platelets, the apoptosis easy occurs in platelets loaded with EGCG. The EGCG accelerates the course of platelet apoptosis.

Healing effect of lyophilized platelets on rat chronic wound model / 中国实验血液学杂志

Platelets carry over 20 growth factors, which all have been shown to improve wound healing, particularly recalcitrant wound healing. The aim of this study was to investigate the healing effect of lyophilized platelets on the chronic wounds through establishing diabetic rat chronic wound model. Healthy male SD rats were intraperitoneally injected with streptozotocin (STZ) solution at the dose of 65 mg/kg. The blood glucose and weights were observed every week. The re-epithelialization rates of normal control group (NDR), diabetic group (DR) and diabetic treatment group (TLP) was analysed. Two full thickness skin wounds were incised in the back of the rats. The re-epithelialization rates were observed at 1, 3, 5, 7 and 12 days. The results showed that after induced by streptozotocin for 72 hours, the blood glucose of the DR group was higher than 16.7 mmol/L. 1 week after induced by STZ, the weight of the DR group was significant lighter than that of the NDR group (p < 0.05). The re-epithelialization rate of DR group were lower than that of NDR. After 12 day treatment, the re-epithelialization rates of NDR and TLP groups were 88.1% and 81.8%, which were significantly higher than that of DR group (62.8%). It is concluded that diabetic rat model established by the intraperitoneal injection of streptozotocin can be used as a better diabetic chronic wound model. And the lyophilized platelets have healing effect on diabetic chronic wounds.

Genetically modified myeloma cell vaccine inducing antitumor immune response in vivo / 中国实验血液学杂志

This study was aimed to evaluate the in vivo antitumor effect of genetically modified myeloma cell vaccine on human myeloma xenografts implanted into nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice. Human immune system was established in NOD/SCID mice by intraperitoneal injection of human peripheral blood lymphocytes (PBLs). After being inoculated subcutaneously with irradiated myeloma cell line sko-007, adenovirally transferred with GFP or p53, granulocyte-macrophage colony-stimulating factor (GM-CSF) and B7-1 genes, huPBL-NOD/SCID mice were challenged by subcutaneous injection of non-transferred sko-007 cells. The results indicated that Ad-p53/GM-CSF/B7-1-infected sko-007 cell vaccination significantly reduced local tumor growth compared with controls. Histopathological and immunohistochemical analysis showed that tumor tissues increasingly displayed diffuse necrosis, mainly caused by apoptosis, accompanied with significant fibroplasias and blood vessel hyperplasia, and human T cells infiltrated into the tumor tissues. It is concluded that transgenic p53, GM-CSF and B7-1 expression produces an immune response against myeloma cells and may be of therapeutic value for multiple myeloma in human being.

Tendency and mathematical model of relationship of blood ATP content with temperature and time of preservation / 中国实验血液学杂志

To investigate the relationship of blood ATP content with temperature and time of preservation and to establish its mathematical model, adenosine triphosphate (ATP) concentration was applied as the index of the quality of erythrocytes; systematical study on variation of blood preserved in a series of different temperatures from 4 degrees C to 32 degrees C was performed, and a series of experimental data were obtained. The results showed that when the ATP concentration y = f (d, t, s) in preserved blood was given as the continuous function of the time (d), the temperature (t) and the initiate ATP concentration (s), the model was fitted with the theory of linearity regression in symbolic statistics, and the general mathematical physical equation of the variation of preserved blood quality was deduced. According to the equation, the whole blood in CPDA-1 solution could be efficiently stored for 35, 35, 29, 22, 18, 18, 13, 8, 7, 6, 6, 5, 4, 4 and 3 days in 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30 and 32 degrees C, respectively. In conclusion, the general tendency of the variation of preserved blood quality according to the temperature and the time was systematically disclosed for the first time, which would be propitious to estimate the blood quality in various temperatures and to instruct clinical blood transfusion.

Labeling and survival studies on rabbit RBC / 中国应用生理学杂志

<p><b>AIM</b>To 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.</p><p><b>METHODS</b>The 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.</p><p><b>RESULTS</b>The 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.</p><p><b>CONCLUSION</b>Compared 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.</p>

Analysis on ultrastructure of lyophilized red blood cells / 中国实验血液学杂志

The objective of the present study was designed to evaluate lyophilized red blood cells of the ultrastructure. Blood was drawn from healthy adult. In group 1, sample was fresh blood; in group 2, sample was added 35% glycerine, stored at -80 degrees C for 24 hours; in group 3, red blood cells stored at 4 degrees C for 5 hours, then were lyophilized for 16 hour. The sample was resuspended for measurements of count and electron microscopy study. The result showed that lyophilized red blood cells possessed relative integrated structure, red blood cell recovery was 53%. The mean diameter, optical density and integral optical density of red blood cell were 4.7 +/- 0.4, 0.14 +/- 0.03 and 1.58 +/- 0.46 in group 1; 4.6 +/- 0.7, 0.14 +/- 0.02 and 2.35 +/- 0.64 in group 2; 4.4 +/- 0.4, 0.17 +/- 0.05 and 2.35 +/- 0.46 in group 3, respectively. There was no significant difference in lyophilized and frozen group, but there was significant difference in lyophilized group and normal group. In conclusion, human red blood cells could be successfully lyophilized and possess relative integrated structure. The mean diameter, optical density and integral optical density of lyophilized red blood cells were similar to that of cryopreservation red cells.

Dynamic changes of ultrastructure of erythrocytes in prolonged preservation at 4 degrees C / 中国实验血液学杂志

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.

New tactics of human red blood cells stored at 4 degrees C-protective effect of antioxidant solution on red blood cells damage / 中国实验血液学杂志

Aliquots of venous blood from healthy donor were collected in plastic blood storage bags with ACD, GMA or antioxidant solution (superoxide dismutase, SOD), respectively, and stored at 4 degrees C. After storage for varying periods, the parameters of the blood were detected in the blood samples. Results showed that the parameters of the blood stored at 4 degrees C for 75 days in SOD group were following: the recovery of RBC-Hb was 87.2%, plasma-Hb (mg/L) was 193.2, P50 (mmHg) was 34.0 (normal value was 33.1); deformability (DImax) was 0.2413 (74.3% of normal value). There was no evident hemolysis, color change, air bubble and clots. It was concluded that human RBC stored at 4 degrees C for 75 days with SOD solution, recovery of levels of RBC-Hb and plasma-Hb were accorded with the requirements of "Basic Demands of Blood Station" in China.

Studies of protective effect of SOD on damage of human RBCs stored at 4 degrees C / 中国应用生理学杂志

<p><b>AIM</b>To 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.</p><p><b>METHODS</b>whole 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.</p><p><b>RESULTS</b>All 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.</p><p><b>CONCLUSION</b>Whole blood in SOD solution can be stored satisfactorily for 75 days at 4 degrees C, and furnished theoretical evidence for RBCs survival.</p>

A simple and convenient method for assaying human RBCs glycolysis rate without oxygen / 中国应用生理学杂志

<p><b>AIM</b>To convenience of the methods for assaying red blood cell glycolysis without oxygen condition in the studies.</p><p><b>METHODS</b>Reagent 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.</p><p><b>RESULTS</b>Human 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%).</p><p><b>CONCLUSION</b>The 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.</p>