Poly （ethylene terephthalate）（dacron, PET） films were exposed under argon plasma glow discharge with different glows and induced polymerization of acrylic acid（AA） in order to in- troduce carboxylic acid group o...Poly （ethylene terephthalate）（dacron, PET） films were exposed under argon plasma glow discharge with different glows and induced polymerization of acrylic acid（AA） in order to in- troduce carboxylic acid group onto PET （PET-AA） assisted by ultraviolet radiation（UV）. Hirudin- immobilized PET （PET-HRD） films were prepared by the grafting of PET-AA, followed by chem- ical reaction with hirudin. The surface structure of the treated PET was determined by X-ray photoelectron spectroscopy （XPS）. The wettability, surface free energy, and interface free energy of the films were investigated by contact angle measurement. The blood compatibility of the films was assessed by platelet-adhesion test and fibrinogen conformational change measurements to eval- uate the viability of the materials in biomedical engineering. Measurement by scanning electron microscopy （SEM） revealed that the amounts of adhered, aggregated and morphologically changed platelets were reduced on the hirudin-immobilized PET films. Enzyme-linked-immunoassay mea- surements that disclosed fibrinogen conformational changes showed results consistent with the platelets＇ behavior.展开更多
Objective: To investigate whether fused hirudin peptide has both antithrombin and antiplatelet functions. Methods: The core region of fused hirudin was the C-terminal tail of hirudin(hirudin_ 53-64),which could bind t...Objective: To investigate whether fused hirudin peptide has both antithrombin and antiplatelet functions. Methods: The core region of fused hirudin was the C-terminal tail of hirudin(hirudin_ 53-64),which could bind to the anion binding exosite (ABE) of thrombin.Arg-Pro-Pro-Gly-Phe(RPPGF) amino acid sequence,a metabolite of bradykinin,was added to the N-terminus of hirudin_ 53-64.It bound to the active site of thrombin.Additionally,Arg-Gly-Asp(RGD)amino acid sequence,an inibitor of glycoprotein Ⅱb/Ⅲa( GP Ⅱb/Ⅲa) receptor,was linked to C-terminus of hirudin_ 53-64.This 26-animo acid-fused hirudin peptide was artificially synthesized,purified and analysed. Results: Fused hirudin peptide significantly lengthened the activated partial thromboplastin time(APTT),thrombin time(TT)and prothrombin time(PT) and inhibited the amidolytic activity of thrombin.The ADP-induced platelet aggregation was markedly inhibited by fused hirudin peptide. Conclusion: Fused hirudin peptide has activity of antithrombin as well as antiplatelet.Therefore bifunctional anticoagulation peptide has capacity to target various components of haemostatic process and may become more powerful antithrombosis agent.展开更多
Objective To investigate distribution and excretion of N-Ile1Thr2-63-desulfatohirudin(rH)a recombinant hirudin newly developed in China,in rats for its development as a novel anticoagulant agent.Methods ELISA was used...Objective To investigate distribution and excretion of N-Ile1Thr2-63-desulfatohirudin(rH)a recombinant hirudin newly developed in China,in rats for its development as a novel anticoagulant agent.Methods ELISA was used to determine the rH concentration in related tissues and body fluids.Tissues were collected at 15,60 and 180min respectively,after iv administration of rH 1.0 mg·kg-1 to 3 groups of 5 rats,and homogenized.Urine,bile and feces were collected at pre-selected intervals of time after iv dosing 1.0 mg·kg-1 to 3 groups of 5 rats and assayed.Results rH following iv dosing was distributed rapidly,the rH levels in all tissues being found to be the highest at 15 min post-injection,afterwards gradually reduced.The highest concentration of rH was found in blood,the next in lung and heart,the lowest in brain.With 15 min post dose as an example,the rH contents in tissues were ranked in order of plasma>lung>heart >adipose>skeletal muscles>kidney>liver>spleen>brain.The 12 h-cumulative excretion amount of rH in urine and feces accounted for 0.03%and 0.001% of administered dose,respectively;the 6 h-cumulative excretion amount in bile was 0.02%of the dose.Conclusions The rH is distributed mainly in blood circulation system with very low content in other tissues.The drug is excreted from urine,feces and bile of rats in extremely minute amount(only 0.051% dose),suggesting that rH undergoes extensive metabolic elimination in rat body.展开更多
目的:探究水蛭素对被动型Heymann肾炎模型大鼠足细胞Nephrin表达的影响。方法:成功建立被动型Heymann肾炎模型,将实验动物分为空白对照组、模型安慰剂组、贝那普利组和水蛭素干预组,每组10只,实验观察周期为4周。于4周后观察大鼠24 h尿...目的:探究水蛭素对被动型Heymann肾炎模型大鼠足细胞Nephrin表达的影响。方法:成功建立被动型Heymann肾炎模型,将实验动物分为空白对照组、模型安慰剂组、贝那普利组和水蛭素干预组,每组10只,实验观察周期为4周。于4周后观察大鼠24 h尿蛋白定量、血肌酐(Scr)、丙氨酸氨基转移酶(ALT)、血小板(PLT)的变化情况,且取出SD大鼠肾脏,采用RT-PCR法测定大鼠肾皮质中Nephrin m RNA的相对表达水平,Western印迹测定Nephrin蛋白的相对表达量,免疫组化法对Nephrin蛋白进行半定量分析。结果:与模型组相比,贝那普利组在Nephrin m RNA和Nephrin蛋白方面差异无统计学意义(P>0.05),但水蛭素干预后可增加Nephrin m RNA和Nephrin蛋白的表达(P<0.05)。相比于贝那普利组,水蛭素组可增加Nephrin m RNA的表达,但在Nephrin蛋白方面差异无统计学意义。免疫组化半定量分析:空白组Nephrin蛋白的表达量最高,贝那普利组、水蛭素组的Nephrin蛋白表达量强于模型组,水蛭素组大鼠肾脏Nephrin蛋白的表达量高于贝那普利组,其差异均有统计学意义(P<0.01)。生化指标显示:与正常组比较,贝那普利组和水蛭素组的24 h尿蛋白、PLT升高,模型组、贝那普利组和水蛭素组Scr显著降低;与模型组比较,贝那普利组的Scr、ALT降低;与贝那普利组比较,水蛭素组Scr偏高。结论:水蛭素可上调被动型Heymann肾炎模型大鼠足细胞Nephrin m RNA和蛋白的表达。展开更多
文摘Poly （ethylene terephthalate）（dacron, PET） films were exposed under argon plasma glow discharge with different glows and induced polymerization of acrylic acid（AA） in order to in- troduce carboxylic acid group onto PET （PET-AA） assisted by ultraviolet radiation（UV）. Hirudin- immobilized PET （PET-HRD） films were prepared by the grafting of PET-AA, followed by chem- ical reaction with hirudin. The surface structure of the treated PET was determined by X-ray photoelectron spectroscopy （XPS）. The wettability, surface free energy, and interface free energy of the films were investigated by contact angle measurement. The blood compatibility of the films was assessed by platelet-adhesion test and fibrinogen conformational change measurements to eval- uate the viability of the materials in biomedical engineering. Measurement by scanning electron microscopy （SEM） revealed that the amounts of adhered, aggregated and morphologically changed platelets were reduced on the hirudin-immobilized PET films. Enzyme-linked-immunoassay mea- surements that disclosed fibrinogen conformational changes showed results consistent with the platelets＇ behavior.
文摘Objective: To investigate whether fused hirudin peptide has both antithrombin and antiplatelet functions. Methods: The core region of fused hirudin was the C-terminal tail of hirudin(hirudin_ 53-64),which could bind to the anion binding exosite (ABE) of thrombin.Arg-Pro-Pro-Gly-Phe(RPPGF) amino acid sequence,a metabolite of bradykinin,was added to the N-terminus of hirudin_ 53-64.It bound to the active site of thrombin.Additionally,Arg-Gly-Asp(RGD)amino acid sequence,an inibitor of glycoprotein Ⅱb/Ⅲa( GP Ⅱb/Ⅲa) receptor,was linked to C-terminus of hirudin_ 53-64.This 26-animo acid-fused hirudin peptide was artificially synthesized,purified and analysed. Results: Fused hirudin peptide significantly lengthened the activated partial thromboplastin time(APTT),thrombin time(TT)and prothrombin time(PT) and inhibited the amidolytic activity of thrombin.The ADP-induced platelet aggregation was markedly inhibited by fused hirudin peptide. Conclusion: Fused hirudin peptide has activity of antithrombin as well as antiplatelet.Therefore bifunctional anticoagulation peptide has capacity to target various components of haemostatic process and may become more powerful antithrombosis agent.
文摘Objective To investigate distribution and excretion of N-Ile1Thr2-63-desulfatohirudin(rH)a recombinant hirudin newly developed in China,in rats for its development as a novel anticoagulant agent.Methods ELISA was used to determine the rH concentration in related tissues and body fluids.Tissues were collected at 15,60 and 180min respectively,after iv administration of rH 1.0 mg·kg-1 to 3 groups of 5 rats,and homogenized.Urine,bile and feces were collected at pre-selected intervals of time after iv dosing 1.0 mg·kg-1 to 3 groups of 5 rats and assayed.Results rH following iv dosing was distributed rapidly,the rH levels in all tissues being found to be the highest at 15 min post-injection,afterwards gradually reduced.The highest concentration of rH was found in blood,the next in lung and heart,the lowest in brain.With 15 min post dose as an example,the rH contents in tissues were ranked in order of plasma>lung>heart >adipose>skeletal muscles>kidney>liver>spleen>brain.The 12 h-cumulative excretion amount of rH in urine and feces accounted for 0.03%and 0.001% of administered dose,respectively;the 6 h-cumulative excretion amount in bile was 0.02%of the dose.Conclusions The rH is distributed mainly in blood circulation system with very low content in other tissues.The drug is excreted from urine,feces and bile of rats in extremely minute amount(only 0.051% dose),suggesting that rH undergoes extensive metabolic elimination in rat body.
文摘目的:探究水蛭素对被动型Heymann肾炎模型大鼠足细胞Nephrin表达的影响。方法:成功建立被动型Heymann肾炎模型,将实验动物分为空白对照组、模型安慰剂组、贝那普利组和水蛭素干预组,每组10只,实验观察周期为4周。于4周后观察大鼠24 h尿蛋白定量、血肌酐(Scr)、丙氨酸氨基转移酶(ALT)、血小板(PLT)的变化情况,且取出SD大鼠肾脏,采用RT-PCR法测定大鼠肾皮质中Nephrin m RNA的相对表达水平,Western印迹测定Nephrin蛋白的相对表达量,免疫组化法对Nephrin蛋白进行半定量分析。结果:与模型组相比,贝那普利组在Nephrin m RNA和Nephrin蛋白方面差异无统计学意义(P>0.05),但水蛭素干预后可增加Nephrin m RNA和Nephrin蛋白的表达(P<0.05)。相比于贝那普利组,水蛭素组可增加Nephrin m RNA的表达,但在Nephrin蛋白方面差异无统计学意义。免疫组化半定量分析:空白组Nephrin蛋白的表达量最高,贝那普利组、水蛭素组的Nephrin蛋白表达量强于模型组,水蛭素组大鼠肾脏Nephrin蛋白的表达量高于贝那普利组,其差异均有统计学意义(P<0.01)。生化指标显示:与正常组比较,贝那普利组和水蛭素组的24 h尿蛋白、PLT升高,模型组、贝那普利组和水蛭素组Scr显著降低;与模型组比较,贝那普利组的Scr、ALT降低;与贝那普利组比较,水蛭素组Scr偏高。结论:水蛭素可上调被动型Heymann肾炎模型大鼠足细胞Nephrin m RNA和蛋白的表达。
