The glycosylation of platelets may prolong their life-span when being transfused after preservation under 4 degrees C, therefore this study was aimed to investigate the effect of glycosylation on morphology, ultrastructure, function and membrane glycoprotein of platelets. The experiments were divided into 3 groups: group preserved in room temperature (RT group), group preserved in 4 degrees C (4T group) and group UDP-Gal glycosylated and preserved in 4 degrees C (U+4T group). The binding rate of RCA I lectin and expression of platelet surface markers CD62P, CD42b were determined by flow cytometry. Morphology and ultrastructure of platelets were observed by light microscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Platelets aggregation was detected by aggregometer. The results showed that the binding rate of RCAI in U+4T group significantly higher than that in RT group (p<0.01), no obvious changes was found in ultrastructure of glycosylated platelets, as compared with fresh platelets. Some morphologic changes, such as pseudopodium could be observed in 4T group. The aggregation rate of platelets in U+4T group reached to 50% of RT group. The expression levels of CD42b and CD62P, and the binding rate of annexin V in U+4T group were not significantly different from that in RT group. It is concluded that UDP-Gal can effectively cause galactosylation of platelets, and the platelets modified with UDP-Gal remain normal morphology, ultrastructure and function.
Blood Platelets ; drug effects ; physiology ; ultrastructure ; Blood Preservation ; methods ; Cryopreservation ; methods ; Galactose ; pharmacology ; Glycosylation ; Humans

This study was aimed to investigate the stability and in vitro function of glycosylated platelets concentrates after long-term refrigeration. The experiments were divided into 4 groups: group preserved at room temperature (RT group), group preserved at 4 degrees C (4T group), group glycosylated and preserved at 4 degrees C (U + 4 group) and group preserved at 4 degrees C and glycosylated (4 + U group). All groups followed for up to 14 days. The binding rate of RCA I lectin and expression of Plt surface markers CD62P, CD42b and Annexin V binding were determined by flow cytometry. pH and mean volume were determined by pH meter and hematotocytometer respectively. Platelet aggregation was detected by aggregometer. The results showed that during storage up to 14 days RCAI binding rate of modified groups was 5 - 6 fold of RT group. The pH of platelets suspension had no significant difference between these two groups (p > 0.05). Mean volumes of both groups (10.6 +/- 1.9 fL and 11.14 +/- 1.1 fL) were also no significant difference (p > 0.05). Furthermore, aggregation responsiveness of modified groups was better than that of RT groups (p < 0.05) although both decreased during the storage. The expression level of CD62P, CD42b and Annexin V binding during 5 days of storage had no significant difference between modified and fresh platelet groups (p > 0.05). While the expression level of CD62P and PS increased and the expression level of CD42b decreased during storage up to 14 days, there was significant difference between modified and fresh platelet groups (p < 0.01). It is concluded that the glycan modification is stable during storage up to 14 days. The glycosylated platelets retain in vitro function better than RT platelets during storage, but it shows activation to varying degrees in vitro after storage for 5 days.
Blood Platelets ; cytology ; metabolism ; Blood Preservation ; Cryopreservation ; methods ; Galactose ; pharmacology ; Glycosylation ; Humans ; Platelet Aggregation ; drug effects

The purpose of this research is to construct a kind of 3D-Scaffold with galactose-carrying polysaccharide for improving the function of hepatocytes in vitro. Galactose moieties were covalently coupled with hyaluronic acid through ethylenediamine. Galactosylated hyaluronic acid/chitosan scaffolds were prepared by lyophilization. The characteristics of the scaffolds such as morphology, hydrophilicity, and mechanical properties were investigated. The results indicated that the porosity and the pore size of the scaffolds made in -20 degrees C were useful used for culturing hepatocytes. And, the incorporating of hyaluronic acid in chitosan network improved the hydrophilicity and mechanical properties of the scaffolds. Rat primary hepatocytes growing in the scaffolds observed by phase-contrast microscope showed the multicellular spheroid morphologies. Therefore, galactosylated hyaluronic acid/chitosan scaffolds could be used as a promising scaffold for liver tissue engineering.
Animals ; Cells, Cultured ; Chitosan ; chemistry ; pharmacology ; Galactose ; chemistry ; pharmacology ; Hepatocytes ; physiology ; ultrastructure ; Hyaluronic Acid ; chemistry ; pharmacology ; Liver ; physiology ; ultrastructure ; Porosity ; Rats ; Tissue Engineering ; Tissue Scaffolds ; chemistry

The study was purposed to develop a novel cryopreserved agent (CPA) for platelets, to investigate the morphology of cryopreserved platelets in different CPA and the CD62P expression on membrane of platelets after stimulating by thrombin, as well as to compare the effect of adding UDP-Gal on preserved efficiency of preservation solutions. A novel cryopreserved agent consisting of 2% DMSO, thrombosol and UDP-Gal was developed on basis of using higher concentration of DMSO. The morphology of chilled platelets was observed by transmission electron microscope and compared with fresh platelets. The expression of CD62P on the membrane of platelets was detected at 0, l, 3 months. The results indicated that the significant effect of cryopreservation on morphology of platelets was found according to percentages of round, dendritic and irregular shapes of cryopreserved platelets. The protective effects of 2% DMSO + thrombosol and 2% DMSO + thrombosol + UDP-Gal were better than that of 5% DMSO. Compared with fresh platelets, the expression of CD62P on platelet membrane decreased obviously after cryopreservation, but not observed difference at preservation for 1 month and 3 months, as well as among 3 kinds of different CPA. It is concluded that the protective effects of 2% DMSO + thrombosol and 2% DMSO + thrombosol + UDP-Gal on morphology of platelets are similar, but better than that 5% DMSO. The reaction of cryopreserved platelets to thrombin decreases, while the significant difference is not found among these 3 kinds of CPA. The addition of UDP-Gal to cryopreserved agents not show the protective effect on platelets.
Blood Platelets ; Blood Preservation ; methods ; Cryopreservation ; methods ; Cryoprotective Agents ; pharmacology ; Dimethyl Sulfoxide ; pharmacology ; Humans ; P-Selectin ; biosynthesis ; genetics ; Uridine Diphosphate Galactose ; pharmacology

OBJECTIVETo investigate the effect of Sijunzi decoction (SJZD) on malondialdehyde (MDA) content and telomerase activity in heart, liver and brain tissues of D-galactose (D-gal) induced aging model mice.

METHODSD-gal aging mice model was established by cervicodorsal region subcutaneous injection with 10% D-gal once a day for six successive months. The model mice in the low-, middle- and high-dose SJZD treated groups were treated with SJZD in a dose of 6 g/kg, 12 g/kg, 24 g/kg per day respectively in the volume of 0.2 ml/10 g for 6 successive weeks. While the mice in the normal control group (NCG, non-modeled) and the model control group (MCG, modeled but untreated) were treated with distilled water instead. The MDA content and telomerase activity in heart, liver and brain tissues of mice was measured with TBA colorimetric method and PCR-ELISA respectively.

RESULTSIn MCG, the MDA content in heart, liver and brain tissues was significantly higher (P < 0.01), and the telomerase activity in liver and heart tissues was significantly lower (P < 0.01) but that in brain tissue was insignificant different to that in NCG (P > 0.05) respectively. As compared with MCG, the MDA content was significantly lower in the three SJZD treated group (P < 0.01). In comparison of telomerase activity between MCG and SJZD treated groups, it was shown that in heart tissue, there was an increased trend of the activity in the low-dose and middle-dose group, but with statistical insignificance (P > 0.05), but it did show a significant increase in the high-dose group (P < 0.05); in liver tissue no significant difference was shown between the three SJZD treated groups and MCG (P > 0.05); as for that in brain tissue, significant increase only shown in the high-dose group (P < 0.01).

CONCLUSIONSJZD can antagonize free radical injury, decrease the MDA content of heart, liver and brain in D-gal aging mice, and increase the telomerase activity in heart and brain tissues but with no effect on that in liver tissue.

Aging ; drug effects ; metabolism ; Animals ; Brain ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Free Radical Scavengers ; pharmacology ; Galactose ; Liver ; metabolism ; Male ; Malondialdehyde ; metabolism ; Mice ; Myocardium ; metabolism ; Telomerase ; metabolism

OBJECTIVETo explore the protective effect of tetramethylpyrazine (TMP) on the learning and memory function in D-galactose (D-gal)-lesioned mice.

METHODSC57BL/6 mice were injected (s.c.) 2% D-gal for 40 days (100 mg x kg(-1) x d(-1)). Normal saline, TMP, and Huperzine A were respectively given by intragastric administration in different groups from the third week. Learning and memory ability was tested with Morris water maze for 5 days at the sixth week. After completion of behavioral test, the mice were sacrificed by decapitation. The brain was rapidly removed, and the cortex and hippocampus were separated. The superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in the cortex were determined. At the same time, the activity of choline acetyltransferase (ChAT) and acetylcholinesterase (AChE), the binding sites (Bmax) and the affinity (KD) of M-cholinergic receptor in the cortex, and Bmax and KD of N-methyl-D-aspartate (NMDA) receptor in the hippocampus were determined.

RESULTSIn this model group, (1) The deficit of learning and memory ability, (2) elevated MDA content and lowered SOD activity, (3) decreased AChE activity and M-cholinergic receptor binding sites in the cortex, and (4) lowered NMDA receptor binding sites were observed in the hippocampus, as compared with the normal control. TMP could markedly (1) attenuate cognitive dysfunction, (2) lower MDA content and elevate SOD activity, (3) increase the activity of ChAT and AChE, and M-cholinergic receptor binding sites in the cortex in the mice treated with D-gal. NMDA receptor binding sites were also increased in the hippocampus in the treated mice.

CONCLUSIONTMP can significantly strengthen antioxidative function, improve central cholinergic system function, protect NMDA receptor activity, and thus enhance the learning and memory ability in D-gal-lesioned mice.

Alzheimer Disease ; chemically induced ; metabolism ; physiopathology ; Animals ; Galactose ; Hippocampus ; metabolism ; Male ; Maze Learning ; drug effects ; Mice ; Mice, Inbred C57BL ; Neuroprotective Agents ; pharmacology ; Pyrazines ; pharmacology ; Receptors, Muscarinic ; metabolism

OBJECTIVETo investigate the inhibiting effects and mechanism of achyranthes bidentata polysaccharide (ABP) and lycium barbarum polysaccharide (LBP) on nonenzyme glycation in D-galactose induced mouse aging model.

METHODSSerum AGE levels were determined by AGE-ELISA, MTT method was used to determine lymphocyte proliferation, IL-2 activity was determined by a bioassay method. Spontaneous motor activity was used to detect mouse's neuromuscular movement, latency of step-through method was used to examine learning and memory abilities of mouse, colormetric assay was used to determine hydroxyproline concentration in mouse skin, pyrogallol autoxidation method was used to determine superoxide dismutase (SOD) activity of erythrocytes.

RESULTSDecreased levels of serum AGE, hydroxyproline concentration in mouse skin and spontaneous motor activity in D-galactose mouse aging model were detected after treated with ABP or LBP, while lymphocyte proliferation and IL-2 activity, learning and memory abilities, SOD activity of erythrocytes, were enhanced.

CONCLUSIONSABP and LBP could inhibit nonenzyme glycation in D-galactose induced mouse aging model in vivo and ABP has a better inhibiting effect than LBP.

Achyranthes ; chemistry ; Aging ; physiology ; Animals ; Disease Models, Animal ; Erythrocytes ; Female ; Galactose ; chemistry ; Learning ; Lycium ; chemistry ; Memory ; Mice ; Mice, Inbred C57BL ; Motor Activity ; Polysaccharides ; pharmacology ; Superoxide Dismutase ; pharmacology

OBJECTIVETo investigate the effects of D-galactose (D-gal) on aging of rat marrow mesenchymal stem cells (MSCs) and its mechanism.

METHODSMSCs isolated from young (7 d) SD rats were randomly divided into four groups:control group, 1g/L, 10g/L and 50g/L D-gal treatment groups. In control group MSCs were cultured in DMEM containing 10% FBS for 48 h. In the D-gal treatment groups, MSCs were cultured in DMEM containing 10% FBS with 1g/L, 10g/L or 50g/L D-gal for 48 h. The senescence-associated changes were examined with SA-β-galactosidase (SA-β-gal) staining, the expressions of p53, p21 and p16 were detected by Western blot. The living and apoptotic cells were determined by AO/EB staining. Cell proliferation was detected by MTT assay. SOD activity was measured by xanthine oxidase method, and the MDA content was estimated with thiobarbituric acid (TBA) method.

RESULTSCompared to control group, the number of SA-β-gal positive cells and the expression of p53, p21 and p16 were significantly increased in the 10g/L and 50g/L D-gal treatment groups. The apoptosis rate in 50g/L D-gal group was significantly higher than that in control group (P<0.01). The proliferation of MSCs was decreased in the 10g/L and 50g/L D-gal groups compared to control group (P<0.05). After 10g/L and 50g/L D-gal treatment, SOD activity was significantly decreased (P<0.01), and MDA level was increased (P<0.01).

CONCLUSIONThe aging of MSCs can be induced by 10g/L and 50g/L D-gal, which may be associated with the elevated levels of oxidative stress.

Animals ; Apoptosis ; drug effects ; Cells, Cultured ; Cellular Senescence ; drug effects ; Galactose ; pharmacology ; Male ; Mesenchymal Stromal Cells ; drug effects ; physiology ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley

The purpose of this paper is to study the effect of kinetin (Kn) on immunity and splenic lymphocyte proliferation in vitro of aging rats induced by D-galactose (D-gal). Fifty SD rats were randomly divided into five groups: control group, aging model group, Kn low dose group, Kn middle dose group and Kn high dose group. The aging model group was proposed by napes subcutaneous injection of D-gal (125 mg/kg) for 45 d, and anti-aging groups were intragastrically administered with 5, 10, 20 mg/kg of Kn respectively from day 11. IgG, IgA, IgM contents of serum, the apoptosis percentage, stimulation index (SI) and proliferation index (PI) of splenic lymphocyte in vitro were evaluated. The results showed that the apoptosis percentage of splenic lymphocyte in aging model rats was higher, the serum IgG, IgA and IgM contents, SI and PI were lower than control group. Kn significantly decreased the apoptosis percentage of splenic lymphocyte, while increased the serum IgG, IgA and IgM contents, SI and PI in aging model group. These results suggest that Kn could inhibit the apoptosis, while promote the proliferation of splenic lymphocyte, and then effectively enhance the immune power of the aging rats and slow down the aging process.
Aging ; drug effects ; immunology ; Animals ; Antibodies ; blood ; Apoptosis ; Cell Proliferation ; drug effects ; Galactose ; adverse effects ; Kinetin ; pharmacology ; Lymphocytes ; cytology ; Rats ; Rats, Sprague-Dawley ; Spleen ; cytology

OBJECTIVETo study the effect of puerarin on fatty superoxide of aged mice induced by D-galactose.

METHODThe aged mice were induced by s.c. 0.12 g.kg-1 D-galactose for 6 weeks. Meanwhile, they were treated with three doses of puerarin once a day for 6 weeks. Then the spontaneous behavior was tested in the aged mice using open field at the next day after the last treatment. And the activity of SOD, the contents of MDA and lipofuscin in brain, liver, and serum were measured with Ultravioletray spectrometer and Fluorospectrophometer analysis system.

RESULTCompared with the D-galactose control group, puerarin of 50 mg.kg-1 and 100 mg.kg-1 puerarin significantly increased the spontaneous behavior, remarkably promoted the activity of SOD of brain, liver, and serum in the aged mice, and significantly decreased the contents of MDA and lipofuscin.

CONCLUSIONPuerarin can improve the activity of antioxidase of the modelling aged-mice induced by D-galactose.

Aging ; blood ; metabolism ; Animals ; Behavior, Animal ; drug effects ; Female ; Galactose ; Isoflavones ; isolation & purification ; pharmacology ; Male ; Malondialdehyde ; metabolism ; Mice ; Plants, Medicinal ; chemistry ; Pueraria ; chemistry ; Superoxide Dismutase ; metabolism