Objective To investigate the influence of NO mixture(L-arginine,apocynin and sodium nitroferricyanide) on wound healing of diabetes mellitus(DM) mice.Methods Fifty adult male Kunming mice were induced by streptozotocin to establish type I DM model.They were then randomly divided into control group,L-arginine group(150 g/L),apocynin group(1?10-4 mol/L),sodium nitroferricyanide group(0.1 mmol/L) and NO mixture group(above components were mixed with equal ratio).Full-thickness skin wound was made and injected with corresponding drugs of 0.15 ml once every 2 days.The wounds were digitally photographed to calculate the percentage of wound closure using computer image analysis software in 1,3,5,7 and 10 d post-injury.The density of fibroblasts,the content of collagenous fibers and the neovascularization in the wound samples were measured with the aid of HE staining.Results From the third day post-injury,the wound healing rate of NO mixture group was significantly higher than that of all the other groups(P

Objective　To evaluate the clinical value of detection of urinary interleukin-8（IL-8） in patients with lupus nephritis （LN）.Methods　Urinary levels of IL-8 were determined by using sandwich ELISA in 38 LN patients.Results　In the LN patients during the　stage　of　normal renal function and diminished renal reserve stage， urinary levels of IL-8 were significantly higher than those in healthy controls（P＜0.001），and urinary levels of IL-8 in the patients during diminished renal reserve stage were significantly higher than those in the patients during normal renal function stage（P＜0.01）.High IL-8 levels in LN patients were associated with high levels of erythrocyte sedimentation rate and lower levels of C3 complement components. Urinary levels of IL-8 in LN patients with microscopic leukocyturia were significantly higher than those without microscopic leukocyturia. In LN patients treated with cyclophosphamide （CTX） and prednisone， urinary IL-8 levels of post-treatment were significantly lower than those of pre-treatment（P＜0.01）. Conclusions 　Urinary levels of IL-8 may be a useful indicator for monitoring disease activity and degree of renal damage in LN. The mechanism of noninfective leukocyturia may be associated with the elevation of urinary IL-8 levels in LN. CTX and prednisone may have a significant effect on inhibiting the production of IL-8 in the immunocytes and renal intrinsic cells.

Polyamidoamine-polyethylene glycol (PAMAM-PEG) copolymers were synthesized using IPDI as coupling reagent by two-step method. The copolymers were characterized by IR spectrum and 1H NMR spectrum, and the PEG conjugating ratios of the copolymers were calculated equal to 10% and 30% separately. MTT assay indicated that after PEGylation a lower cytotoxicity of the copolymers could be found, and with increasing PEG conjugating ratio the cytotoxicity decreased obviously. Agarose gel retardation assay demonstrated that PAMAM-PEG copolymers could be combined with DNA and PAMAM-PEG/DNA complexes were prepared by self-assembly. DLS measurement showed that when N/P > or = 50, the particle size of copolymer/ gene complexes was in a range of 150-200 nm, and the zeta potential was in a range of 10-25 mV. In vitro gene transfection illustrated that when N/P < or = 50, the gene transfection efficiency of PAMAM-PEG copolymers was a little less than that of PAMAM-G5, but the transfection efficiency can be raised by increasing N/P ratio or transfection time. Considering both cytotoxicity and transfection efficiency aspects PAMAM-PEG-13 was more effect than PAMAM-PEG-39 in PEGylation.

BACKGROUND: At present, after transplantation of small diameter artificial blood vessel, long-term patency rate is low due to being lacking of endothelial cells for lining and anti-thrombus characters. In some studies,mature endothelial cells were tried to be seeded in the artificial vessel to boost up its anti-thrombus capability so as to improve the long-term patency rate, but we got unsatisfied effect due to the defects of seed cells and scaffolds. Therefore, in clinic, proper seed cells and vascular scaffolds have been searched for improving the long-term low pateney rate in transplantation of small diameter artificial blood vessel.OBJECTIVE: To investigate the feasibility that differentiation of bone marrow mononuclear cells induced in vitro into endothelial-progenitor cells (EPCs) and seed polyurethane small diameter artificial blood vessel so as to provide proper seed cells for endotheliazation of polyurethane small diameter artificial blood vessel.DESIGN: Observation experiment SETTING: Cardivascular Medical Department and Staff Room of Immunology, First Hospital Affiliated to Sun Yat-sen University MATERILAS: This experiment was carried out at the First Hospital Affiliated to Sun Yat-sen University from September 2004 to May 2005. About 10 mL of bone marrow from healthy adult volunteers (n=7) was used in this experiment.METHODS: Bone marrow mononuclear cells of healthy adult were collected and put in the fibronectin pre-coated DMEM culture medium, then induced by vascular endothelial growth factor and basic fibroblast growth factor. Induced cells were observed under fluorescence microscope and identified with immunohistochemical staining. The induced and proliferated EPCs were seeded onto the surface of polyurethane small diameter artificial blood vessel. Morphological change was observed under scanning electron microscope.MAIN OUTCOME MEASURES: ① Cellular morphological change.② Staining results of immunohistochemical VWF and CD 34 antibody . ③ Adhesive growth status of EPCs on the polyurethane small diameter artificial blood vessel RESULTS: ① In the vascular endothelial growth factor and basic fibroblast growth factor and other inducers , bone marrow mononuclear cells differentiated into EPCs , presenting typical "spindle-shaped" appearance under an inverted fluorescence microscope and became to form a monolayer that arrayed in "cobblestone-like" ② Immunohistochemical staining showed von willebrand factor(VWF) and CD34 antigen stained positive. ③ Under the scanning electron microscope, surface of polyurethane small diameter artificial blood vessel without seeded cells presented typical polyporous honeycomb-like structure , and the size of hole suited the crawling of EPCs. After seeding the cells, we observed the adhesion, crawling and spreading of the EPCs on the surface of polyurethane small diameter artificial blood vessel. Some EPCs grew into the honeycomb-like holes were seen occasionally.CONCLUSION: Bone marrow mononuclear cells can be induced and differentiated into EPCs, while induced and differentiated EPCs well grow adhesively in the polyurethane small diameter artificial vessels, suggesting that differentiation of bone marrow mononuclear cells induced in vitro into EPCS, which can be used as seed cells for endothelialization of polyurethane small diameter artificial blood vessels.

In order to investigate the ability of PEG-PEI copolymers as gene carriers for delivery of VEGF165. A series of PEG-PEI copolymers with different PEG grafting was prepared and the cytotoxicity was evaluated. Simultaneously,the VEGF165 gene segment with HindⅢ and BamHⅠ site was obtained by PCR, which was cloned into pEGFP-C1. PEG-PEI/ pEGFP-VEGF165 complexes were formed by self-assembly and transfected HUVEc. Transfection efficiency was evaluated by measuring the percentage of cells expressing green fluorecensce protein. The VEGF expression was detected by ELISA, RT-PCR, and the effect of transfection on growth of endothelial cell was evaluated by MTT. The results suggested that the formation of PEG-PEI copolymers could help to reduce the cytotoxicity of PEI. After transfection, the strong expression of green fluorescence protein was observed by fluorescence microscopy. The transfection efficiency was influenced by the number of PEG side chains and N/P ratio. Of all copolymers tested, the transfection efficiency of PEG-PEI(5-25-1) at N/P = 30 reached a maximum, which was much higher than that of PEI. The expression of VEGF protein and mRNA increased significantly, and HUVEc proliferation was accelerated after transfection.These results indicates PEG-PEI copolymers can be used as effective gene carriers for delivery of pEGFP-VEGF165 gene.

Objective To explore the effects of secreted apoptosis-related protein 1 (SARP1) on apoptosis of the hyperstrophic scar fibroblasts (HSFB) and its regulating mechnisms.Methods The recombinant vector was identified by enzyme digestion analysis.And the virus supernatant of the recombinant vector was extracted from packaged 293 cells,then it infected the skin fibroblasts from hypertrophic scar patients,which aimed to promote its expression of SARP1 protein.After adenovirus infection,the expression of SARP1 in the fibroblasts was confirmed by RT-PCR and Western blot.The effect of SARP1 on proliferation of HSFB was detected by MTT assay,and the effect of SARP1 on apoptosis of HSFB was detected and change of the cells functions were analyzed by FACS.Results Recombinants were confirmed.After adenovirus infection,both protein and mRNA of SARP1 were detected in HSFB.And the mRNA value of SARPlwas detected to increase significantly by RT-PCR and the protein expression was detected to increase significantly by Western blot (P＜0.05).The proliferation in the groups of the adenovirus infection and HSFB was positively regulated by SARP1 (P＜0.01) and the apoptosis of them was inhibited by the expression of SARP1 as compared to the control groups of HSFB and Ad-EGFP.It showed that the apoptosis index decreased as compared the group of infected fibroblasts to the control group by flow cytometry.Conclusions SARP1 could be highly expressed in HSFB by adenovirus infection,exhibiting the proliferation-enhancing and apoptosis-inhibiting effects on HSFB.

The radical compliance of small diameter artificial vascular grafts was measured by a device made of laser micrometer, pressure transducer, A/D card, micro computer and pulsed circulation loop, the volumetric compliance was measured by a device made of micro-syringe and pressure transducer; the longitudinal compliance was calculated from volumetric compliance and radical compliance. The research results showed that the radical compliance, volumetric compliance and longitudinal compliance would rise not only with the increase in polyurethane (PU) elasticity and salt/polymer ratio (or porosity), but also with the decrease in dipping layers (or wall thickness). Circumferential moduli E zeta and longitudinal moduli E theta could be calculated from volumetric compliance and longitudinal compliance respectively; E theta and E zeta would decrease with the increase in PU elasticity and salt/polymer ratio, but was independent on number of dipping layers. Small diameter PU artificial vascular grafts with compliance close to natural vein or artery can be prepared by choosing of more elastic PU materials (Chro or PCU1500), optimization of salt/polymer ratio (6:1), and the number of dipping layers (4-6 layers).
Biocompatible Materials ; chemistry ; Blood Vessel Prosthesis ; Compliance ; Elasticity ; Humans ; Polyurethanes ; chemistry

Copoly (benzyl glutamate-hydroxy ethyl glutamine) were prepared by partially aminolysis of poly benzyl glutamate with hydroxyl ethylamine at 60 degrees C for predeternined period. As aminolysis was proceeding, the content of hydroxy ethyl glutamine in copolymer increased and the yield of aminolysis decreased. When aminolysis time reached 16.5 hours, a copolymer with 0.357 mole fraction of hydroxy ethyl glutamine was obtained. After 16.5 hours a soluble yellow viscous product of reaction was obtained. The aminolysized specimens displayed more swelling degree in water, that is, more hydrophilits. The meassurement of solid-liquid contact angles showed that an increase in critical surface tention with content of hydroxy ethyl glutamine in copolymer was observed. Likewise a obvious increase in polar component and a slight decrease in dispersive of surface free energy, thus an increase in total surface free energy with content of hydroxy ethyl glutamine was found. The platelet adhesion and deformation test indicated that less platelets were adhered to surfaces of all aminolysized specimens than that of both poly benzyl glutamate and polydimethylsilicone. The least adhered platelets on surface of copolymer with 0.133 mole fraction of content of hydroxy ethyl glutamine were observed. The partiall aminolysis of poly benzyl glutamate films is an effective method to improve its hydrophility and antithrombogenicity.
Animals ; In Vitro Techniques ; Male ; Materials Testing ; Platelet Adhesiveness ; Polyglutamic Acid ; analogs & derivatives ; chemistry ; Rabbits ; Surface Properties

OBJECTIVEThe -455 G/A (HaeIII) polymorphism of beta-fibrinogen gene influences levels of plasma fibrinogen. We further investigated whether it influences the risk of ischemic cerebrovascular disease.

METHODSWe accumulated 134 acute ischemic cerebrovascular disease (ICVD) cases and compared their -455 G/A status with a control group (n = 166). The beta-fibrinogen gene -455 G/A polymorphism was analyzed for all subjects by PCR-RFLP with the restrictive enzyme HaeIII.

RESULTSPlasma fibrinogen was higher in AA homozygous participants (341 mg/dL) than in participants carrying the G allele: GA (290 mg/dL), GG (298 mg/dL) in the control group. Plasma fibrinogen was also higher in AA homozygous patients (353 mg/dL) than in cases carrying the G allele: GA (287 mg/dL), GG (302 mg/dL) in the ICVD group. However, there was no significant association between beta-fibrinogen gene -455 G/A polymorphism and ICVD group.

CONCLUSIONSAlthough a small effect cannot be excluded, beta-fibrinogen gene -455 G/A polymorphism is an independent predictor of plasma fibrinogen, but not of ischemic cerebrovascular disease.

Alleles ; Cerebrovascular Disorders ; blood ; genetics ; Female ; Fibrinogen ; genetics ; metabolism ; Genetic Predisposition to Disease ; Homozygote ; Humans ; Male ; Middle Aged ; Polymorphism, Restriction Fragment Length ; Risk Factors

The blood compatibility of block copolymer membranes of poly(benzyl L-glutamate)/poly(ethylene glycol) and the effect of on the blood compatibility of copolymer were evaluated by the clotting time test, the platelet adhesion and deformation test, and the protein adsorption test. The results showed that in terms of blood compatibility, homopolymer was better than glass and silicone, copolymer was better than homopolymer, and the more the PEG in the copolymer, the better the blood compatibility.
Biocompatible Materials ; Drug Carriers ; Materials Testing ; Platelet Adhesiveness ; drug effects ; Polyethylene Glycols ; chemical synthesis ; chemistry ; Polyglutamic Acid ; analogs & derivatives ; chemical synthesis ; chemistry ; Polymers ; chemical synthesis ; chemistry