Two different immunoassay methods, competitive indirect enzyme-linked immuno-sorbent assay (CI-ELISA) and amplificative competitive indirect ELISA (ACI-ELISA) using biotin-avidin complex system were studied to detect ...Two different immunoassay methods, competitive indirect enzyme-linked immuno-sorbent assay (CI-ELISA) and amplificative competitive indirect ELISA (ACI-ELISA) using biotin-avidin complex system were studied to detect rhEPO. The linear ranges were 50-20000 ng/mL and 10-50000 ng/mL for CI-ELISA and ACI-ELISA, respectively. The low detection limits of CI-ELISA and ACI-ELISA were 62.8 ng/mL and 8.5 ng/mL, respectively.展开更多
To assess the efficacy and the optimum dose of recombinant human erythropoietin (rhEpo) on the anemia of premature, 45 preterm infants with a gestational age of less than 35 weeks and birth weight of less 1 800 g were...To assess the efficacy and the optimum dose of recombinant human erythropoietin (rhEpo) on the anemia of premature, 45 preterm infants with a gestational age of less than 35 weeks and birth weight of less 1 800 g were randomly assigned to treatment group 1 (n = 15, receiving subcutaneous rhEpo 150 U/kg·time), treatment group 2 (n = 15, receiving 250 U/kg·time), three times a week for 6 weeks, and control group (n = 15, no treatment was given). All preterm infants received supplements of vitamin E (20 IU) and iron (20 mg) each day. Our results showed that postnatal decline of hemoglobin (Hb) and hematocrit (Hct) were lessened in the treatment groups, particularly in the group 2 and the differences were very significant (P<0. 0001 for all). Treated infants had significantly higher reticulocyte counts (Ret) (P<0. 000] for all), but there was no significant difference between the two treatment groups (P>0. 05). Serum iron dropped significantly in the treatment groups as compared with control group (P<0. 01 for all), but no dose-dependent relationship was observed in treated infants (P>0. 05). After treatment, serum levels of erythropoietin was higher in group 2 than those in group 1 and control group (P<0. 0001, P<0. 01 and P<0. 05, respectively). There was no significant difference between group 1 and control group (P>0. 05). No side effects related to rhEpo therapy were observed. Our study suggested that rhEpo therapy stimulates endogenous erythro-poiesis and enhances Ret, Hct and level of Hb in a dose-dependent manner in premature infants. The therapy is more efficient when given in higher dosages.展开更多
BACKGROUND Hema-Plus,a recombinant human erythropoietin(rHuEPO)or epoetin alfa has shown effectiveness in correction of anemia in Thai population in clinical practice.This study was aimed to demonstrate efficacy and s...BACKGROUND Hema-Plus,a recombinant human erythropoietin(rHuEPO)or epoetin alfa has shown effectiveness in correction of anemia in Thai population in clinical practice.This study was aimed to demonstrate efficacy and safety under the evidencebased approach.AIM To evaluate the efficacy and safety of rHuEPO(Hema-Plus)for treatment of anemia over 12 wk in Thai patients with Stage V chronic kidney disease(CKD)on peritoneal dialysis(PD).METHODS This study was an open-label,multi-center study to enroll 30 CKD patients identified to start PD with hemoglobin(Hb)less than 9.5 g/dL,serum ferritin more than 100 ng/mL,serum transferrin saturation more than or equal to 20%and who had not previously received epoetin.Patients with conditions that could increase the risk of adverse effects from study participation or interfere with study outcomes,were using concomitant androgens or had secondary hyperparathyroidism were excluded.All eligible patients started Hema-Plus by SC injection at 4000 IU once or twice weekly(week 0)and with follow-up at weeks 2,4,8,and 12.Dosage adjustment could be done to achieve Hb level of 11-12 g/dL.Primary end point was mean change in Hb level from baseline to end of treatment(week 12).Safety was assessed throughout the study.Quality of life(QoL)was assessed using KDQOL-36.RESULTS All 30 enrolled patients completed the study.Mean(standard deviation)Hb at baseline(week 0)to the end of 12 wk was significantly increased from 7.39(1.29)g/dL to 11.15(1.73)g/dL(paired t-test,P value<0.001).Overall change of Hb means from baseline over the other 4 visits was statistically significantly increased(repeated measure ANOVA,P value<0.001).Ten out of 39 adverse events(AEs)were serious.Two serious AEs were probably related to study medication by investigators’assessment.At week 12,the QoL scores in all domains were significantly increased from baseline.CONCLUSION Hema-Plus administered for 12 wk for treatment of anemia in patients on PD effectively increased Hb levels with acceptable safety profile.展开更多
Objective To produce specific monoclonal antibody （mAb） against recombinant human erythropoietin （rHuEPO） for development of highly efficient methods for erythropoietin detection in biological fluids. Methods rHuE...Objective To produce specific monoclonal antibody （mAb） against recombinant human erythropoietin （rHuEPO） for development of highly efficient methods for erythropoietin detection in biological fluids. Methods rHuEPO was covalently coupled with bovine serum albumin （BSA） and the conjugate was used to immunize mice to produce specific mAb against rHuEPO based on hybridoma technology. The obtained F3-mAb was characterized by enzyme-linked irmnunosorbent assay （ELISA）, SDS-PAGE and Western blot. Results The isotype of F3-mAb was found to be IgM with an affinity constant of 2.1x10s L/mol. The competitive ELISA using the obtained IgM showed a broader linear range and lower detection limit compared with previous work. Conclusions The modification of rHuEPO was proved to be successful in generating required specific mAb with high avidity to rHuEPO.展开更多
Objective To observe the expression of hypoxia inducible factor-1α (HIF-1α) in the retina of rabbits with acute high intraocular pressure and to investigate the mechanism of systemic domestic recombinant human eryth...Objective To observe the expression of hypoxia inducible factor-1α (HIF-1α) in the retina of rabbits with acute high intraocular pressure and to investigate the mechanism of systemic domestic recombinant human erythropoietin (rhEPO) protecting the retina from ischemia-reperfusion injury. Methods First,control group and model group were established in rabbit eyes. The acute high intraocular pressure model was established by saline perfusion into anterior chamber,and then hypodermic injection of domestic rhEPO was made. HIF-1α protein in the retina was observed by immunohistochemical staining method on days 1,3,7 and 14 after retinal ischemia-reperfusion,respectively. Results No cells with HIF-1α positive expression were observed in the retina of the control group. Cells with HIF-1α positive expression in the model group outnumbered those in the control group (P<0.01). The resemblance pattern occurred in EPO group but its degree was slightly greater than that in the model group from day 3 after ischemia-reperfusion (P<0.05). Conclusion Domestic rhEPO can down-regulate the expression of HIF-1α in the retina with acute high intraocular pressure,which may be one of the mechanisms that rhEPO protects the retina from ischemia-reperfusion injury.展开更多
Objective: To explore the regulative effect on Survivin of r-HuEPO after traumatic brain injury (TBI) in rats, and understand the neuroprotection mechanisms of r-HuEPO. Methods: Seventy-eight adult Wistar rats were ra...Objective: To explore the regulative effect on Survivin of r-HuEPO after traumatic brain injury (TBI) in rats, and understand the neuroprotection mechanisms of r-HuEPO. Methods: Seventy-eight adult Wistar rats were randomly divided into sham operation group (n = 6), TBI group (n = 36) and r-HuEPO group (n = 36). The experimental TBI model was created by Feeney’s method. Samples were obtained after injury for measuring apoptosis of cells by Epics XL Flow Cytometer. Immunochemical method was performed for inspection of expressions of Survivin and NF-κB proteins. Results: Compared to the sham group, the number of apoptotic cells and Survivin, NF-κB immunopositive cells was significantly increased in the injured brain after TBI (P < 0.01). R-HuEPO significantly increased the expression of survivin and NF-κB, but decreased the apoptotic rates. Conclusion: Increased expression of NF-κB by r-HuEPO may play important role in regulating Survivin level, inhibiting neuronal apoptosis in cortex and exerting protective function to neurons.展开更多
AIM:Anaemia caused by acute upper gastrointestinal bleeding is treated with blood transfusion or iron, but patients usually face a two-month recovery period from post-haemorrhage anaemia. This prospective, randomised,...AIM:Anaemia caused by acute upper gastrointestinal bleeding is treated with blood transfusion or iron, but patients usually face a two-month recovery period from post-haemorrhage anaemia. This prospective, randomised, open,pilot study was designed to investigate whether recombinant human erythropoietin (Epoetin) therapy accelerate haematocrit increase in the post-bleeding recovery period.METHODS:We studied hospitalised patients admitted because of acute ulcer bleeding or haemorrhagic gastritis,who had a haematocrit of 27-33% and did not receive blood transfusions.One day alter the endoscopic confirmation of cessation of bleeding, they were randomised either to erythropoietin (20000 IU Epoetin alfa subcutaneously, on days 0, 4 and 6) plus iron (100mg im, on days 1-6, (G1) or iron only (G2). Haematocrit was measured on days 0, 6, 14,30, 45, and 60, respectively.R.ESULTS: One patient from G1 and two from G2 were lost to follow-up. Therefore, 14 and 13 patients from G1 and G2 respectively were analysed. Demographic characteristics, serum iron, ferritin, total iron binding capacity, reticulocytes, and haematocrit were not significantly different at entry to the study.Median reticulocyte counts were significantly different between groups on day six (G1: 4.0,3.0-6.4 vs G2:3.5,2.1-4.4%,P=0.03) and median haematocrit on day fourteen [G1:35.9,30.7-41.0 vs G2:32.5,29.5-37.0% (median, range), P=0.04].CONCLUSION:Erythropoietin administration significantly accelerates correction of anemia alter acute ulcer bleeding.The haematocrit gain is equivalent to one unit of transfused blood two weeks alter the bleeding episode.展开更多
以EPO抗体修饰的Fe_(3)O_(4)粉末-聚丙烯酰胺凝胶为结合相,明胶凝胶为扩散层,通过薄膜扩散梯度技术(DGT)与紫外光谱法,对去离子水、人体尿液中的rhEPO进行富集和检测的实验研究。在25℃、常压环境下,DGT中结合相的饱和富集容量为0.0758...以EPO抗体修饰的Fe_(3)O_(4)粉末-聚丙烯酰胺凝胶为结合相,明胶凝胶为扩散层,通过薄膜扩散梯度技术(DGT)与紫外光谱法,对去离子水、人体尿液中的rhEPO进行富集和检测的实验研究。在25℃、常压环境下,DGT中结合相的饱和富集容量为0.0758μg cm^(2),当pH值为5.5~8.0、无机盐浓度为0.3%~1.5%时,rhEPO的检测回收率基本不受影响。对质量浓度为6~15 ng mL的rhEPO水溶液进行检测时,rhEPO检测回收率的平均值为81.9%;对质量浓度为5~20 ng mL的rhEPO人体尿液进行检测时,rhEPO检测回收率的平均值为79.8%。展开更多
目的探讨重组人促红细胞生成素(recombinant human erythropoietin,rhEPO)对6-羟基多巴胺(6-OHDA)诱导的SD大鼠帕金森病(PD)模型小胶质细胞活化的影响。方法 40只SD大鼠随机分为A组(rhEPO+6-OHDA)、B组(生理盐水+6-OHDA)、C组(6-OHDA)、...目的探讨重组人促红细胞生成素(recombinant human erythropoietin,rhEPO)对6-羟基多巴胺(6-OHDA)诱导的SD大鼠帕金森病(PD)模型小胶质细胞活化的影响。方法 40只SD大鼠随机分为A组(rhEPO+6-OHDA)、B组(生理盐水+6-OHDA)、C组(6-OHDA)、D组(生理盐水),每组10只。(1)A组:右侧纹状体内立体定向注射重组促红细胞生成素(rhEPO),24h后同侧黒质内立体定向注射6-OHDA;(2)B组:右侧纹状体内立体定向注射与rhEPO等量的生理盐水,24h后同侧黒质内立体定向注射6-OHDA;(3)C组:右侧黒质内立体定向注射6-OHDA;(4)D组:右侧黒质内立体定向注射与6-OHDA等量的生理盐水。4w后采用免疫组化检测黒质内酪氨酸羟化酶(TH)阳性神经元和CD11b阳性细胞数量及CD11b阳性细胞形态变化。结果与D组比较,A组大鼠黒质TH阳性神经元明显减少,CD11b阳性细胞明显增多,大部分小胶质细胞胞体小,突起细长;与B组和C组比较,A组大鼠黒质TH阳性神经元显著增多,CD11b阳性细胞显著减少,仅有少量小胶质细胞胞体大,突起短粗。结论重组人促红细胞生成素(rhEPO)可能通过抑制小胶质细胞活化,减轻6-OHDA对多巴胺(DA)能神经元的毒性损害,对DA能神经元产生神经保护作用。展开更多
文摘Two different immunoassay methods, competitive indirect enzyme-linked immuno-sorbent assay (CI-ELISA) and amplificative competitive indirect ELISA (ACI-ELISA) using biotin-avidin complex system were studied to detect rhEPO. The linear ranges were 50-20000 ng/mL and 10-50000 ng/mL for CI-ELISA and ACI-ELISA, respectively. The low detection limits of CI-ELISA and ACI-ELISA were 62.8 ng/mL and 8.5 ng/mL, respectively.
文摘To assess the efficacy and the optimum dose of recombinant human erythropoietin (rhEpo) on the anemia of premature, 45 preterm infants with a gestational age of less than 35 weeks and birth weight of less 1 800 g were randomly assigned to treatment group 1 (n = 15, receiving subcutaneous rhEpo 150 U/kg·time), treatment group 2 (n = 15, receiving 250 U/kg·time), three times a week for 6 weeks, and control group (n = 15, no treatment was given). All preterm infants received supplements of vitamin E (20 IU) and iron (20 mg) each day. Our results showed that postnatal decline of hemoglobin (Hb) and hematocrit (Hct) were lessened in the treatment groups, particularly in the group 2 and the differences were very significant (P<0. 0001 for all). Treated infants had significantly higher reticulocyte counts (Ret) (P<0. 000] for all), but there was no significant difference between the two treatment groups (P>0. 05). Serum iron dropped significantly in the treatment groups as compared with control group (P<0. 01 for all), but no dose-dependent relationship was observed in treated infants (P>0. 05). After treatment, serum levels of erythropoietin was higher in group 2 than those in group 1 and control group (P<0. 0001, P<0. 01 and P<0. 05, respectively). There was no significant difference between group 1 and control group (P>0. 05). No side effects related to rhEpo therapy were observed. Our study suggested that rhEpo therapy stimulates endogenous erythro-poiesis and enhances Ret, Hct and level of Hb in a dose-dependent manner in premature infants. The therapy is more efficient when given in higher dosages.
基金the Ethics Committee,Institute for the Development of Human Research Protections(IHRP),No.IHRP 2365/2556The Ethics Committee of Udon Thani Hospital,No.8/2559and Siriraj Institutional Review Board Faculty of Medicine Siriraj Hospital,Mahidol University,No.Si 588/2016.
文摘BACKGROUND Hema-Plus,a recombinant human erythropoietin(rHuEPO)or epoetin alfa has shown effectiveness in correction of anemia in Thai population in clinical practice.This study was aimed to demonstrate efficacy and safety under the evidencebased approach.AIM To evaluate the efficacy and safety of rHuEPO(Hema-Plus)for treatment of anemia over 12 wk in Thai patients with Stage V chronic kidney disease(CKD)on peritoneal dialysis(PD).METHODS This study was an open-label,multi-center study to enroll 30 CKD patients identified to start PD with hemoglobin(Hb)less than 9.5 g/dL,serum ferritin more than 100 ng/mL,serum transferrin saturation more than or equal to 20%and who had not previously received epoetin.Patients with conditions that could increase the risk of adverse effects from study participation or interfere with study outcomes,were using concomitant androgens or had secondary hyperparathyroidism were excluded.All eligible patients started Hema-Plus by SC injection at 4000 IU once or twice weekly(week 0)and with follow-up at weeks 2,4,8,and 12.Dosage adjustment could be done to achieve Hb level of 11-12 g/dL.Primary end point was mean change in Hb level from baseline to end of treatment(week 12).Safety was assessed throughout the study.Quality of life(QoL)was assessed using KDQOL-36.RESULTS All 30 enrolled patients completed the study.Mean(standard deviation)Hb at baseline(week 0)to the end of 12 wk was significantly increased from 7.39(1.29)g/dL to 11.15(1.73)g/dL(paired t-test,P value<0.001).Overall change of Hb means from baseline over the other 4 visits was statistically significantly increased(repeated measure ANOVA,P value<0.001).Ten out of 39 adverse events(AEs)were serious.Two serious AEs were probably related to study medication by investigators’assessment.At week 12,the QoL scores in all domains were significantly increased from baseline.CONCLUSION Hema-Plus administered for 12 wk for treatment of anemia in patients on PD effectively increased Hb levels with acceptable safety profile.
基金This work was supported by the National Natural Science Foundation of China (No.20675006).
文摘Objective To produce specific monoclonal antibody （mAb） against recombinant human erythropoietin （rHuEPO） for development of highly efficient methods for erythropoietin detection in biological fluids. Methods rHuEPO was covalently coupled with bovine serum albumin （BSA） and the conjugate was used to immunize mice to produce specific mAb against rHuEPO based on hybridoma technology. The obtained F3-mAb was characterized by enzyme-linked irmnunosorbent assay （ELISA）, SDS-PAGE and Western blot. Results The isotype of F3-mAb was found to be IgM with an affinity constant of 2.1x10s L/mol. The competitive ELISA using the obtained IgM showed a broader linear range and lower detection limit compared with previous work. Conclusions The modification of rHuEPO was proved to be successful in generating required specific mAb with high avidity to rHuEPO.
基金supported by the Science and Technology Research Foundation of Shaanxi Province, China (No.2008K14-07)the Medical and Phar macologic Research Foundation of Xi'an City, China (No.SF08004-2)
文摘Objective To observe the expression of hypoxia inducible factor-1α (HIF-1α) in the retina of rabbits with acute high intraocular pressure and to investigate the mechanism of systemic domestic recombinant human erythropoietin (rhEPO) protecting the retina from ischemia-reperfusion injury. Methods First,control group and model group were established in rabbit eyes. The acute high intraocular pressure model was established by saline perfusion into anterior chamber,and then hypodermic injection of domestic rhEPO was made. HIF-1α protein in the retina was observed by immunohistochemical staining method on days 1,3,7 and 14 after retinal ischemia-reperfusion,respectively. Results No cells with HIF-1α positive expression were observed in the retina of the control group. Cells with HIF-1α positive expression in the model group outnumbered those in the control group (P<0.01). The resemblance pattern occurred in EPO group but its degree was slightly greater than that in the model group from day 3 after ischemia-reperfusion (P<0.05). Conclusion Domestic rhEPO can down-regulate the expression of HIF-1α in the retina with acute high intraocular pressure,which may be one of the mechanisms that rhEPO protects the retina from ischemia-reperfusion injury.
文摘Objective: To explore the regulative effect on Survivin of r-HuEPO after traumatic brain injury (TBI) in rats, and understand the neuroprotection mechanisms of r-HuEPO. Methods: Seventy-eight adult Wistar rats were randomly divided into sham operation group (n = 6), TBI group (n = 36) and r-HuEPO group (n = 36). The experimental TBI model was created by Feeney’s method. Samples were obtained after injury for measuring apoptosis of cells by Epics XL Flow Cytometer. Immunochemical method was performed for inspection of expressions of Survivin and NF-κB proteins. Results: Compared to the sham group, the number of apoptotic cells and Survivin, NF-κB immunopositive cells was significantly increased in the injured brain after TBI (P < 0.01). R-HuEPO significantly increased the expression of survivin and NF-κB, but decreased the apoptotic rates. Conclusion: Increased expression of NF-κB by r-HuEPO may play important role in regulating Survivin level, inhibiting neuronal apoptosis in cortex and exerting protective function to neurons.
文摘AIM:Anaemia caused by acute upper gastrointestinal bleeding is treated with blood transfusion or iron, but patients usually face a two-month recovery period from post-haemorrhage anaemia. This prospective, randomised, open,pilot study was designed to investigate whether recombinant human erythropoietin (Epoetin) therapy accelerate haematocrit increase in the post-bleeding recovery period.METHODS:We studied hospitalised patients admitted because of acute ulcer bleeding or haemorrhagic gastritis,who had a haematocrit of 27-33% and did not receive blood transfusions.One day alter the endoscopic confirmation of cessation of bleeding, they were randomised either to erythropoietin (20000 IU Epoetin alfa subcutaneously, on days 0, 4 and 6) plus iron (100mg im, on days 1-6, (G1) or iron only (G2). Haematocrit was measured on days 0, 6, 14,30, 45, and 60, respectively.R.ESULTS: One patient from G1 and two from G2 were lost to follow-up. Therefore, 14 and 13 patients from G1 and G2 respectively were analysed. Demographic characteristics, serum iron, ferritin, total iron binding capacity, reticulocytes, and haematocrit were not significantly different at entry to the study.Median reticulocyte counts were significantly different between groups on day six (G1: 4.0,3.0-6.4 vs G2:3.5,2.1-4.4%,P=0.03) and median haematocrit on day fourteen [G1:35.9,30.7-41.0 vs G2:32.5,29.5-37.0% (median, range), P=0.04].CONCLUSION:Erythropoietin administration significantly accelerates correction of anemia alter acute ulcer bleeding.The haematocrit gain is equivalent to one unit of transfused blood two weeks alter the bleeding episode.
文摘以EPO抗体修饰的Fe_(3)O_(4)粉末-聚丙烯酰胺凝胶为结合相,明胶凝胶为扩散层,通过薄膜扩散梯度技术(DGT)与紫外光谱法,对去离子水、人体尿液中的rhEPO进行富集和检测的实验研究。在25℃、常压环境下,DGT中结合相的饱和富集容量为0.0758μg cm^(2),当pH值为5.5~8.0、无机盐浓度为0.3%~1.5%时,rhEPO的检测回收率基本不受影响。对质量浓度为6~15 ng mL的rhEPO水溶液进行检测时,rhEPO检测回收率的平均值为81.9%;对质量浓度为5~20 ng mL的rhEPO人体尿液进行检测时,rhEPO检测回收率的平均值为79.8%。
文摘目的探讨重组人促红细胞生成素(recombinant human erythropoietin,rhEPO)对6-羟基多巴胺(6-OHDA)诱导的SD大鼠帕金森病(PD)模型小胶质细胞活化的影响。方法 40只SD大鼠随机分为A组(rhEPO+6-OHDA)、B组(生理盐水+6-OHDA)、C组(6-OHDA)、D组(生理盐水),每组10只。(1)A组:右侧纹状体内立体定向注射重组促红细胞生成素(rhEPO),24h后同侧黒质内立体定向注射6-OHDA;(2)B组:右侧纹状体内立体定向注射与rhEPO等量的生理盐水,24h后同侧黒质内立体定向注射6-OHDA;(3)C组:右侧黒质内立体定向注射6-OHDA;(4)D组:右侧黒质内立体定向注射与6-OHDA等量的生理盐水。4w后采用免疫组化检测黒质内酪氨酸羟化酶(TH)阳性神经元和CD11b阳性细胞数量及CD11b阳性细胞形态变化。结果与D组比较,A组大鼠黒质TH阳性神经元明显减少,CD11b阳性细胞明显增多,大部分小胶质细胞胞体小,突起细长;与B组和C组比较,A组大鼠黒质TH阳性神经元显著增多,CD11b阳性细胞显著减少,仅有少量小胶质细胞胞体大,突起短粗。结论重组人促红细胞生成素(rhEPO)可能通过抑制小胶质细胞活化,减轻6-OHDA对多巴胺(DA)能神经元的毒性损害,对DA能神经元产生神经保护作用。
