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【Objective】 To observe the effect of glutaraldehyde polymerized bovine hemoglobin injection (code: HSRP1) oxygen-carrying fluid on early perfusion of severe hemorrhagic anemia in rabbits. 【Methods】 The rabbit model of controlled severe hemorrhagic anemia was established. Twelve modeled rabbits were divided into glutaraldehyde polymerized bovine hemoglobin injection (code: HSRP1) group and sodium lactate ringer′s injection (LR) group, with 6 rabbits in each group(half male and half female). HSPR1 group and LR group were treated with HSRP1 and LR, respectively. The survival rate of experimental rabbits was observed, and the indexes of hemodynamics, venous blood gas, plasma hemoglobin, base surplus, lactic acid and bicarbonate were measured before and after blood loss, as well as each point within 24 h after infusion. 【Results】 The survival rate of HSRP1 group was significantly different from that of LR group (P<0.05); After exsanguination, the mean arterial pressure of each group was significantly different from that before exsanguination (P<0.05), but there was no significant difference between HSPR1 group and LR group after infusion; In the second stage of perfusion, the blood lactate concentration and base excess in the HSRP1 group were significantly different from those in the LR group at each time point (P<0.05), at 2 h after perfusion, the respiratory rate started to differ significantly from that of the LR group (P<0.05), heart rate was significantly different from LR group at 4 h after perfusion(P<0.05); There were no significant differences between HSRP1 group and LR group in plasma venous oxygen partial pressure, venous oxygen saturation and plasma hemoglobin at all time points. 【Conclusion】 HSPR1 is used for severe traumatic hemorrhagic shock in rabbits, and can improve the survival rate of experimental rabbits by providing oxygen to hypoxic tissues and correcting anaerobic metabolism. As a new oxygen-carrying fluid, HSPR1 can correct the oxygen supply balance of patients with severe hemorrhagicanemia in early stage.
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A novel fluorescent imprinted polymer (CDs@ MIP) with selective recognition of hemoglobin was prepared by the sol-gel method using fluorescent carbon dots as the carrier material, 3-aminopropyltrieth-oxysilane as the functional monomer, tetraethoxysilane as the crosslinking agent and bovine hemoglobin as template molecule. The results of IR and scanning electron microscopy showed that the molecularly imprinted polymer was coated on the surface of fluorescent carbon dots. The CDs@ MIP showed selective recognition properties for bovine hemoglobin with an imprinting factor of 4. 60. Also the adsorption ability and specific recognition performance of CDs@ MIP were investigated, and it was found that the CDs@ MIP had high selectivity toward bovine hemoglobin, and the selection factors for ovalbumin, bovine serum albumin and human serum albumin were 4. 38, 4. 73 and 3. 66, respectively. Under the optimal conditions, the linear range of CDs@ MIP for bovine hemoglobin was 0. 1-10. 0 μmol/ L and the detection limit was 23. 0 nmol/ L. The CDs@ MIP was successfully used for the determination of bovine hemoglobin in bovine blood samples with recoveries of 99. 0％ -102. 5％ .
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OBJECTIVE: To investigate the toxicity of N-succinyl-chitosan (NSCS) to bovine hemoglobin (BHb) and human liver cells (HL-7702). METHODS: BHb was used as a research object and the toxic effect of NSCS was investigated by UV-Vis absorption spectroscopy, fluorescence spectroscopy and synchrotron spectroscopy under the simulative human physiological condition. At the same time, human HL-7702 cells was used as a research object and methyl thiazolyl tetrazolium (MTT) assay was employed to examine the cytotoxicity of NSCS. RESULTS: The results of UV-Vis absorption spectroscopy and MTT showed that the toxicity of NSCS was weak, and substitution degree had little effect on it. The result of fluorescence spectroscopy demonstrated that the intrinsic fluorescence of BHb was quenched by NSCS and the quenching effect slightly increased with the increase of substitution degree. The quenching mechanism was mainly dynamic quenching, and the major driving forces were hydrophobic and electrostatic force. CONCLUSION: The result of synchronous fluorescence spectroscopy reveals that NSCS has almost no influence on the conformation of BHb. The toxicity of NSCS to BHb and HL-7702 is weak.
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OBJECTIVE: To study the reaction yield of SC-mPEG with bovine hemoglobin(BHB) under different conditions so as to establish a method for the analysis of the SC-mPEG-BHB mixtures.METHODS: The product was analyzed with the methods of SDS-PAGE,GPC,CE,TNBS,etc.The optimal reaction system for SC-mPEG with BHB was established based on the effects of different factors on SC-mPEG-modified BHB.RESULTS: The optimum reaction conditions for SC-mPEG with BHB were established as follows: SC-mPEG and BHB were mixed at an optimal ratio of 60∶1 and the reaction was lasted for 2 h in(pH9 and 4 ℃) buffer solution.CONCLUSIONS: This study established a foundation for the following study of red blood cell substitutes.
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OBJECTIVE: Researching the appropriate methods to separation,purification and detection the mixture of PEG modifier SC-mPEG with BHB.METHODS: Adopting cation-exchange chromatography and organic solvent precipitation to separate the mixture,then detecting it by SDS-PAGE and TNBS methods.RESULTS: Modifer and non-modifer product were separation by cation-exchange chromatography.The average modification rates of two peak product from cation-exchange chromatography are 52.95% and 14.19% detected by TNBS methods.CONCLUSIONS: Cation-exchange chromatography methods could be to separate and purify the SC-mPEG-BHB,TNBS methods coule be to detect the average modification rate of the SC-mPEG-BHB.
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Objective To evaluate the protective effects of stroma-free bovine hemoglobin(SFBHB)on preservation of the donor heart.Methods 24 isolated perfused rabbit hearts were equally divided into two groups.In control group,the cardiac arrest were induced with St Thomas Ⅱ solution and repeated every 30 min.the myocardial temperature remained at 40℃.In experimental group,the isolated rabbit hearts were perfused continuously with St Thomas Ⅱ solution containing stroma-free bovine hemoglobin and the myocardial temperature remained at 37℃.The preservation lasted 6 h.Results There were significant differences between the two groups in the follows:(1)The mean values of the left ventricular developed pressure(LVDP)and coronary flow rate(CFR)were higher in experimental group(P＜0.001);(2)The ATP level and myocardial oxygen uptake were significantly decreased in control group(P＜0.001);(3)The mean values of released creatine phosphate kinase(CPK),lactec dehydrogenase(LDH)and myocardial contents of malondialdehyde(MDA)were higher in control group(P＜0.01);(4)More water was present in the myocardial in control group(P＜0.01);(5)The myocardial ultrastructural injuries were greater in control group.Conclusion The stroma-free bovine hemoglobin could offer great protective effects on preservation of the isolated rabbit hearts.