Effects of recombinant human erythropoietin on cerebral blood flow in preterm infants: arterial spin labeling magnetic resonance imaging evaluation / 中国新生儿科杂志

Objective:To study the effects of recombinant human erythropoietin (rhEPO) on cerebral blood flow (CBF) in preterm infants using arterial spin labeling (ASL) magnetic resonance imaging (MRI).Methods:From September 2021 to June 2022, preterm infants (gestational age ≤32 weeks, birth weight ≤1 500 g) admitted to NICU of our hospital within 24 h after birth were randomly assigned into rhEPO group and control group for this prospective study. The rhEPO group was given rhEPO (500 IU/kg iv, once every other day for 2 weeks) within 72 h after birth plus symptomatic supportive treatment. The control group received same amount of normal saline injection. Both groups received brain MRI, diffusion-weighted imaging and ASL at adjusted gestational age of 35~37 weeks and CBF values of interested areas were measured.Results:A total of 85 infants were enrolled, including 40 in the rhEPO group and 45 in the control group. No significant differences existed in the incidences of periventricular-intraventricular hemorrhage, periventricular leukomalacia, focal white matter injury and extensive white matter injury between the two groups ( P>0.05). The CBF values [ml/(100 g·min)] of frontal cortex [left 15.1±3.9 vs. 17.9±3.1, right 15.9 (12.5, 17.8) vs. 18.1(16.1,20.2)], temporal cortex [left 15.8±4.3 vs. 18.6±3.8, right 16.3(13.2,19.4) vs. 18.1(15.7,19.7)], occipital cortex (left 15.8±6.1 vs. 18.8±3.3, right 16.8±5.5 vs. 19.3±4.8), basal ganglia (left 24.7±7.2 vs. 28.7±6.2, right 26.0±7.9 vs. 29.3±6.4) and thalamus (left 32.7±11.8 vs. 37.9±8.6, right 32.1±11.6 vs. 37.6±10.2) in the rhEPO group were significantly lower than the control group ( P<0.05). No significant differences existed of CBF value at the parietal cortex between the two groups ( P>0.05). Conclusions:Early application of rhEPO can reduce CBF in premature infants, which may be related to the neuro-protective effects of EPO.

Effect of neuroprotective agent assisted with mild hypothermia on curative efficacy and laboratory indexes of hypoxic-ischemic encephalopathy children / 中国中西医结合急救杂志

Objective To observe the effect of recombinant human erythropoietin injection (RHEI) assisted with mild hypothermia on the clinical efficacy and safety of children with hypoxic-ischemic encephalopathy (HIE). Methods From January 2015 to December 2017, 110 children with HIE were treated in Shijiazhuang No.1 Hospital. Fifty-five children with routine treatment were taken as Western medicine routine treatment group. In addition, 55 children treated with mild hypothermia combined with RHEI were taken as mild hypothermia+RHEI group. Both groups were treated for 14 days and followed up for 10 months. The neonatal behavioral neurological assessment (NBNA) score, mental development index (MDI), psychomotor development index (PDI), myelin basic protein (MBP), S100B protein and neuron specific enolization enzyme (NSE), nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), insulin growth factor-1 (IGF-1), growth hormone (GH), and differences in clinical efficacy in two groups were compared, and the occurrence of adverse reactions was observed. Results The NBNA, MDI, PDI, NGF (μg/L), BDNF (ng/L), IGF-1 (pg/L), and GH (pg/L) of two groups after treatment were higher than those before treatment (the Western medicine routine treatment group: 33.72±3.19 vs. 26.81±2.38, 78.95±5.51 vs. 71.39±4.24, 79.62±4.93 vs. 71.84±4.15, 123.74±22.98 vs. 104.29±15.36, 1 518.35±174.92 vs. 1 197.28±148.43, 38.25±4.96 vs. 23.16±2.87, 39.27±5.24 vs. 20.97±3.15; the mild hypothermia+RHEI group: 39.82±3.36 vs. 26.78±2.53, 84.13±6.29 vs. 71.34±4.27, 85.26±5.74 vs. 71.88±4.13, 145.28±27.52 vs. 104.72±15.41, 1 925.71±204.37 vs. 1 192.61±150.26, 57.94±6.62 vs. 23.13±2.91, 56.43±7.14 vs. 20.94±3.17), NSE (μg/L), MBP (μg/L) and S100B (μg/L) were lower than those before treatment (the Western medicine routine treatment group: 17.05±2.26 vs. 24.96±2.83, 9.71±1.85 vs. 23.14±3.37, 0.93±0.12 vs. 1.49±0.24; the mild hypothermia+RHEI group:12.48±1.94 vs. 25.03±2.81, 5.48±1.42 vs. 23.17±3.35, 0.61±0.07 vs. 1.51±0.25). After treatment, the changes of each index in the mild hypothermia+RHEI group were more significant than those in the control group [NABA:39.82±3.36 vs. 33.72±3.19, MDI: 84.13±6.29 vs. 78.95±5.51, PDI: 85.26±5.74 vs. 79.62±4.93, NSE (μg/L):12.48±1.94 vs. 17.05±2.26, MBP (μg/L): 5.48±1.42 vs. 9.71±1.85, S100B (μg/L): 0.61±0.07 vs. 0.93±0.12, NGF (μg/L): 145.28±27.52 vs. 123.74±22.98, BDNF (ng/L): 1 925.71±204.37 vs. 1 518.35±174.92, IGF-1 (pg/L):57.94±6.62 vs. 38.25±4.96, GH (pg/L): 56.43±7.14 vs. 39.27±5.24, all P < 0.05]. The total effective rate of mild hypothermia+RHEI group was significantly higher than that of Western medicine routine treatment group [94.55% (52/55) vs. 81.82% (45/55), P < 0.05]. There were no serious adverse reactions in the two groups. Conclusion RHEI assisted with mild hypothermia therapy can significantly improve the clinical efficacy and NBNA, MDI, PDI scores of HIE children, reduce the degree of brain injury, and improve the neurological function, with good safety.

Prolonged response to recombinant human erythropoietin treatment in patients with myelodysplastic syndrome at a single referral centre in Brazil

OBJECTIVES: To evaluate the effects of epoetin (EPO) alfa treatment on overall survival, event-free survival and response duration in patients with myelodysplastic syndrome (MDS) who were treated at a haematological referral centre in northeastern Brazil. METHODS: This was a retrospective cohort study of 36 patients diagnosed with MDS and treated with EPO alfa at 30,000 to 60,000 IU per week. Clinical data were collected from medical records. The events assessed were non-response to treatment and progression to acute myeloid leukaemia (AML). Statistical analyses were performed using GraphPad Prism 7 and SPSS 24 software. RESULTS: The overall survival of patients who received EPO alfa treatment was 51.64%, with a median of 65 months of treatment, and the overall survival of this group was 100% during the first 24 months. We detected a 43.5-month median event-free survival, with a response rate of 80.5%. We observed responses from 25 to 175 months. Patients with transfusion dependence and those with a high-risk stratification, as determined by the International Prognostic Scoring System (IPSS), the Revised International Prognostic Scoring System (IPSS-R), the WHO classification-based Prognostic Scoring System (WPSS) and the WHO 2016, had a lower event-free survival than other patients. CONCLUSIONS: Despite the wide use of EPO alfa in the treatment of anaemia in patients with MDS, the median response duration is approximately only 24 months. Our data provide encouraging results concerning the benefits of using EPO alfa for the improvement of the quality of life, as patients treated with EPO showed higher overall survival, event-free survival rates and longer response durations than have been previously described in the literature.

Clinical study of recombinant human erythropoietin combined with iron to correct perioperative anemia in elderly patients with intertrochanteric fractures / 中国修复重建外科杂志

Objective : To investigate the effectiveness of recombinant human erythropoietin (rHuEPO) combined with iron in treatment of anemia in elderly patients with intertrochanteric fractures during perioperative period. Methods: A clinical data of 71 patients with intertrochanteric fractures met the inclusion criteria between April 2016 and October 2017 was retrospectively analyzed. All patients were treated with closed reduction and proximal femoral intramedullary nail fixation. Thirty-one patients were treated with rHuEPO and iron before operation as trial group, and 40 patients were not treated with rHuEPO and iron as control group. There was no significant difference in gender, age, body mass index, cause of injury, fracture side and classification, American Society of Anesthesiologists (ASA) classification, combined medical diseases, time from fracture to admission, preoperative hospital stay, and operation time between the two groups ( P>0.05).The hemoglobin levels before operation and at 1, 3, and 7 days after operation, number of blood transfusion, blood transfusion rate, blood transfusion volume, postoperative hospital stay, complications were recorded and compared. Results: After operation, 8 patients (25.8%) in trial group and 22 patients (55.0%) in control group received blood transfusion; the blood transfusion volume was (1.96±0.85) units in trial group and (3.19±1.61) units in control group. There were significant differences in blood transfusion rate and volume between the two groups ( P0.05). The postoperative hemoglobin level was higher in trial group than in control group, and the difference between the two groups was significant at 7 days ( P0.05). All patients were discharged from hospital normally, and no one died during hospitalization. Conclusion: The application of rHuEPO combined with iron before operation in elderly patients with intertrochanteric fractures can rapidly increase the hemoglobin level after operation, shorten the hospital stay, and do not increase the risk of deep venous thrombosis after operation.

Effects of recombinant human erythropoietin and β-aescin on functional recovery and secondary injury of nerve cells in rats after spinal cord injury / 中国免疫学杂志

Objective:To study the effect of recombinant human erythropoietin and β-aescin on functional recovery and secondary injury of nerve cells in rats after spinal cord injury.Methods:A total of 50 female SD rats were randomly divided into five groups according to their injury and treatment.The rats in the rh-EPO group were treated with recombinant human erythropoietin,and the β-SE group was treated with β-aescin,and the combination group was treated with recombinant human erythropoietin and the treatment of β-aescin,compared the indicators.Results: The BBB motor function scores of the combination therapy group were significantly higher than those of rh-EPO group,β-SE group and injury group.The neurological function of the combined treatment group was better than that of rh-EPO group and β-SE group(P<0.05).The content of Mg2+in the combination group was significantly higher than that in the rhEPO group and the β-SE group,and the difference was significant (P<0.05).Conclusion: The combination of re-combinant human erythropoietin and β-aescin can help improve the BBB motor function score,promote the recovery of motor function and delay the development of secondary injury of nerve cells.

Estimation of the Degree of Salivary Acidification of Recombinant-Human-Erythropoietin(rhEPO) / 中国药学杂志

OBJECTIVE: To detect and analyze the degree of salivary acidification of rhEPO isoforms. METHODS: The isoelectric points of rhEPO isoforms were determined with full column imaging capillary isoelectric focusing electrophoresis. And the charge distribution among rhEPO isoforms was analyzed. The degrees of rhEPO's total saliva acidification were measured using the method of appendices of Chinese Pharmacopoeia. At last, the degrees of saliva acidification of rhEPO isoforms were obtained using multivariate linear fitting. RESULTS: Nine kinds of rhEPO isoforms were distinguished and defined as isoform 1 to 9 with isoelectric points in the range of 3.6 to 5.1. There was one sialic acid molecule between two contiguous rhEPO isoform. Furthermore, the degrees of salivary acidification of the main four kinds of isoforms, 4-7, were 13, 12, 11 and 10 mol/mol, respectively. CONCLUSION :This study lays foundation for rhEPO biosimilar evaluation and further analysis of each isoform of rhEPO.

Literature Analysis of Adverse Reactions Induced by Recombinant Human Erythropoietin in 149 Case Re-ports / 中国药师

Objective:To study the adverse reactions (ADRs) of recombinant human erythropoietin (rHuEPO) to provide refer-ence for clinical rational and safe medication. Methods:ADRs induced by rHuEPO reported at home and abroad were collected and analyzed in respects of age,gender,original illness, occurrence time, clinical manifestations and the results. Results: After the re-trieval,there were 149 cases of rHuEPO-induced ADRs with the damage of cardio vascular system, hematologic system, skin and its appendents accounting for 43.4%,20.8% and 12.7%,respectively. The top three main clinical manifestations of rHuEPO drug reac-tions were hypertension,pure red-cell aplastic anemia (PRCA) and hyperkalemia. The occurrence time should be paid particular at-tention in 5-12 weeks after the administration (43.0%). Conclusion:Physicians should be aware of rHuEPO-induced ADRs (espe-cially the occurrence time),pay attention to patients' medication education and avoid serious adverse reactions.

Effects of rHu-EPO in Different Dose on Brain Injury of Premature Infants / 医学研究杂志

Objective To study the effects and side effects of different dose of rHu-EPO on the treatment of brain injury in premature infants. Methods The infants who had suffered fetal distress and first one minute of Apgar was under 7score after birth and were sent to NICU within 24h were studied. We totally collected 90 infants and divided into three groups randomly, including large dose group, small dose group and control group. The large and small group were injected hypodermicly with rHu-EPO 1000U/kg, 500U/kg, three times per week for three weeks, and the control group were given general treatment without rHu-EPO at same time. Before treatment, one week and three weeks after treatment, we collected concentration of NSE, S100B and skull ultrasound to assess the effects. Neonatal behavioral neurological assessment(NBNA) were performed twice before and at weeks of correct gestational age. To survey the side effects, we collected general information such as the incidence rate of ROP and hemangioma, AST/ALT/PLT/Urea /Cr and so on. Results After one-week treatment, the concentration of NSE and S100B were no significant difference(P ＞ 0. 05) in the small dose group, but were statistically significant in the large dose group(P ＜ 0. 05). After three-week treatment, the comparison of NSE、S100B in both groups was statistically significant(P ＜ 0. 05). The head ultrasound comparison was of statistically significant in both group(P ＜ 0. 05), and so as NBNA and head MRI. The frequency of blood transfusion was statistically significant in both group(P ＜ 0. 05) compared with control group. Routine blood test including liver and kidney function showed that there was no significant difference before and after treatment(P ＞ 0. 05). Conclusion The neuroprotective effect of rHu-EPO on brain injury in preterm infants is connected with its dose and period of treatment, it need high dose or long time to express neuroprotective function.

Observation of clinical effects of recombinant human erythropoietin combined with methylprednisolone sodium succinate in the treatment of acute spinal cord injury / 中国综合临床

Objective To evaluate the effect of recombinant human erythropoietin (rhEPO) combined with methylprednisolone sodium succinate (MPSS),compared to MPSS alone,in the treatment of neurological function of patients with acute spinal cord injury (SCI).Methods Twenty-one patients presenting in hospital within less than 8 hours after acute SCI were randomly divided into two groups,the control group (10 cases) and the intervention group (11 cases).The control group was treated by MPSS combined with placebo,while the intervention group received MPSS with rhEPO.Both groups received MPSS 30 mg/kg within the first hour,and if the patient was admitted within 4 hours,MPSS would be applied in the treatment with 5.4 mg/kg per hour in the subsequent 23 hours and till 47 hours if the patient was admitted within 4-8 hours after injury.The intervention group received 500 U/kg rhEPO on admission and another 500 U/kg in the next 24 hours,compared with the control group where placebo was used.The evaluation on neurologic function recovery was made on admission,24 h,72 h,one week,2 months and 6 months later,and statistical analysis was performed.Results The change in ASIA score: in the control group,the increase was seen from admission to 6 months after injury in terms of exercise,algesia and tactile sensation ((31.2±6.6) points vs.(57.8±9.8) points,(41.4±9.5) points vs.(64.3±10.6) points, (39.2±6.8) points vs,(61.5±11.3) points),the increase also took place in the intervention group ((29.5±7.2) points vs.(77.4±10.3) points,(39.7±7.2) points vs.(82.3±12.1) points,(37.4±6.2) points vs.(78.6±12.4) points).As time went on,the increase range in the intervention group became larger,compared with the control group.The difference between the two groups in ASLA score was statistically significant (P0.05).Conclusion The application of MPSS combined with rhEPO within 8 hours after acute spinal injury may be more effective than MPSS with placebo in the neurologic dysfunction recovery.

Effects of exogenous recombinant human erythropoietin on neuronal apoptosis and 5-lipoxygenase in neonatal rats with hyperoxia brain injury / 中华实用儿科临床杂志

Objective To explore the effect of exogenous recombinant human erythropoietin (rhEPO) on neuronal apoptosis in neonatal rats after hyperoxia brain injury.Methods Thirty neonatal Wistar rats were randomly divided into 3 groups by random number table method:rhEPO treatment + 800 mL/L hyperoxia group (group A),9 g/L saline +800 mL/L hyperoxia group (group B),9 g/L saline + air group (group C).Group A was given subcutaneous injection of rhEPO 1 000 IU/kg for 5 days.Group B and group C received the same dose of 9 g/L saline.Group A and group B were continuously exposed to atmospheric pressure hyperoxia model cabin to maintain the oxygen concentration in the container (800 ± 30) mL/L for 5 days.During the course of the experiment,the general situation and weight changes in rats were observed.After 5 d,all rats were sacrificed and brain tissues were taken.Neuronal apoptosis in hippocampal structural region of the newborn rats was observed by terminal deoxynucleotidyl transferase dUTP nick and labeling(TUNEL) staining.Immunohistochemical method was used to detect the expression of 5-lipoxygenase in hippocampal structural region of newborn rats.Results The weight gain and brain weight of group B were lower than those of group C,the weight gain and brain weight of group A were higher than those of group B,and the differences were statistically significant(F =11.179,8.140,all P ＜ 0.05).In group A and group B were found that the neuronal nucleus of the hippocampal neurons was partially contracted,deeply dyed,and the neuronal arrangement was loose,even with local neuron deletions and focal necrosis,but in group A neuron density was higher with less necrosis than that in group B.The neuronal cells in hippocampal structural region were neat and intact in group C.The number of TUNEL positive cells in hippocampal structural region of group B[(6.20 ± 1.93) number/high power field] was significantly higher than that in group C [(1.80 ± 0.79) number/high power field],the number of TUNEL positive cells in hippocampal structural region of group A [(4.20 ± 1.32) number/high power field] was significantly lower than that in group B,and the difference was statistically significant (F =23.912,P ＜ 0.05).The number of 5-lipoxygenase positive cells in group B [(6.90 ± 1.29) number/high power field] was significantly higher than that in group C [(1.00 ± 0.67) number/high power field],the number of 5-lipoxygenase positive cells in group A [(5.60 ± 0.97)number/high power field] was significantly lower than that in group B,and the difference was statistically significant (F =95.044,P ＜ 0.05).Conclusion rhEPO has a protective effect on neonatal rats with hyperoxia brain injury,and alleviates brain cell apoptosis caused by hyperoxia brain injury,which may interfere with the 5-lipoxygenase pathway.

Effects of recombinant human erythropoietin β injection on levels of SOD, GSH-PX, MDA and Hcy with diabetic peritoneal dialysis / 中国生化药物杂志

Objective To investigate the effects of recombinant human erythropoietin βinjection on levels of superoxide dismutase ( SOD ) , glutathione peroxidase ( GSH-PX ) , malondialdehyde ( MDA ) and homocysteine ( Hcy ) in patients with diabetic peritoneal dialysis.Methods 92 patients of parallel peritoneal dialysis in diabetic nephropathy who received therapy from September 2014 to September 2016 in our hospital were selected and randomly divided into the observation group and the control group with 46 cases in each group.The control group was treated with peritoneal dialysis routine treatment, while the observation group was treated with recombinant human erythropoietin βinjection on this basis.The levels of hemoglobin (Hb), hematocrit (Hct), renal function, SOD, GSH-PX, MDA and Hcy were compared.Results After treatment, the levels of Hb and Hct in the observation group were higher than the control group, the difference was statistically significant (P<0.05), the urinary albumin excretion rate (UAER) and serum creatinine (SCr) in the observation group were lower than the control group, the difference was statistically significant (P<0.05), the levels of SOD and GSH-PX in the observation group were higher than the control group, the levels of MDA and Hcy were lower the control group, the difference was statistically significant (P<0.05).Conclusion The effect of recombinant human erythropoietin βinjection on diabetic nephropathy patients with peritoneal dialysis was significant, which could improve the levels of SOD, GSH-PX, MDA and Hcy.

Comparison of primary structures of recombinant human erythropoietin marketed in China / 中国药学杂志

OBJECTIVE: To characterize and compare the primary structures of recombinant human erythropoietin marketed in China. METHODS: Recombinant human erythropoietin reference substances were obtained from nine manufacturers, for which the molecular mass, peptide mass mapping, oligosaccharide profile and content of sialic acid were analyzed and compared. RESULTS: The measured molecular masses of de-N-glycosylated EPO were all in agreement with the theoretical values with mass error less than 1. All the samples had consistent amino acid sequence, disulfide bonds (Cys7-Cys161, Cys29-Cys33), O-glycosylation site (Ser126), and N-glycosylation sites (Asn24, Asn38, Asn83), but different glycosylation pattern and ratio of glycoforms. The sialic acid content of the nine samples were within 11.5-15.7 mol∶mol EPO. CONCLUSION: Recombinant human erythropoietins from nine manufacturers have identical primary structures except for glycosylation patterns.

Potency Evaluation of Recombinant Human Erythropoietin in Brazil: Assessment of Reproducibility Using a Practical Approach

In this study, we compared the results of potency determination of recombinant human erythropoietin (rhEPO) obtained between 2010 and 2012 by the National Institute of Quality Control in Health (INCQS/Fiocruz), i.e., the National Control Laboratory (NCL), and by a manufacturer of rhEPO. In total, 47 different batches of commercially prepared rhEPO (alpha isoform) were analyzed. All results, including those of the control and warning limits, remained within the limits recommended by European Pharmacopoeia (Ph. Eur.). All relative error (RE) values were less than ± 30%, wh ereas most were approximately ± 20%. Applying the Bland-Altman plot, only two of 47 values remained outside the limits of agreement (LA). In addition, agreement of potency determination between INCQS and the manufacturer coefficient of variation of reproducibility (% CVR) was considered satisfactory. Taken together, our results demonstrate (i.) the potency assay of rhEPO performed at INCQS, is standardized and controlled, (ii.) the comparison of our results with those of the manufacturer, revealed an adequate inter-laboratory variation, and (iii.) the critical appraisal proposed here appears to be a feasible tool to assess the reproducibility of biological activity, providing additional information regarding monitoring and production consistency to manufacturers and NCLs.

Clinical effect of recombinant human erythropoietin on patients with primary brain stem injury / 中国综合临床

Objective To observe the short-term curative effect and safety of recombinant human erythropoietin(rHu-EPO)on patients with primary brain stem injury. Methods Sixty patients with primary brain stem injury were recruited at Liaocheng People' Hospital from July 2010 to July 2013. All cases were randomly divided into EPO group and control group. The patients in EPO group were injected subcutaneous with rHu-EPO five times at dose of 6 000 U,while patients in the control group were treated with placebo in 2 weeks. All other conventional treatments were the same. NIHSS score and GOS score were evaluated in two weeks and three months respectively. Moreover,blood pressure and hemoglobin were also measured. Results NIHSS score in EPO group was 11. 37 ± 7. 78,significant higher than that of control group after two weeks(19. 41 ± 8. 26,P = 0. 019). GOS score in EPO group was also significant differences in two groups after three months (Z = - 2. 367,P = 0. 009 ). However,no significant difference was observed in the followed-up blood tests. Conclusion Recombinant human erythropoietin could be the exact nerve protective effect,and might be an effective therapy for patients with primary brain stem injury.

The influence of rHuEPO on inflammatory factors in the process of renal interstitial fibrosis and its possible mechanism / 医学研究生学报

Objective Clinical treatment can delay the development of renal interstitial fibrosis , but it can not reverse renal dysfuntion.The article was to discuss the influence of recombinant human erythropoietin ( rHuEPO ) on inflammatory factors in the process of renal interstitial fibrosis and its possible mechanism . Methods The vitro cultured HK-2 cells were randomized into 7 groups:the blank control group , rHuEPO control group ( addition of 20U/mL rHuEPO), albumin stimulation group (addition of 5mg/mL albumin), 5mg/mL rHuEPO intervention group (5mg/mL albumin +5U/mL rHuEPO), 10 U/mL rHuEPO intervention group (5mg/mL albumin +10 U/mL rHuEPO), 20U/mL rHuEPO intervention group (5mg/mL albumin +20U/mL rHuEPO), and Rho inhibi-taion group (addition of 5mg/mL albumin 30min after 10μmol/L Y27632), 24 h acting time for each group.We observed the changes of cell morphology in each group .Reverse transcription polymerase chain reaction ( RT-PCR) was used to evaluate the mRNA levels of RhoA, ROCK1 and IL-6 , and ELISA was applied to measure the levels of supernatant TNF-αand IL-6 protein. Results The form of pebbles or paving stone was observed in blank control group and rHuEPO intervention groups , a long and thin spindle change with the appearance of fibre cells in albumin stimulation group , the transformation to pebbles in 5, 10, 20 mg/mL rHuEPO intervention groups , the form of oval and slightly increased intercellular space in Rho inhibitaion group .Compared with the blank control group , the expressions of RhoA mRNA, ROCK1 mRNA and IL-6 mRNA significantly increased in the albumin stimulation group (P<0.05), while significantly reduced in 5, 10, 20 mg/mL rHuEPO intervention groups (P<0.05), which was in negative relation with the rHuEPO concentrations .Compared with the albumin stimulation group , the expressions of ROCK 1 mRNA and IL-6 mRNA reduced in Rho inhibtation group (P<0.05), while there was no significant difference as to the expression of RhoA mRNA .ELISA results showed:compared with blank control group , the expressions of supernatant TNF-α([452.32 ±33.23] ng/L vs [1347.54 ±41.52] ng/L), IL-6 protein([884.62 ±0.73] pg/L vs [95.12 ±0.32]pg/LP<0.05) increased significantly.Compared with albumin stim-ulation group, the expressions of TNF-αin 5, 10, 20 mg/mL rHuEPO intervention groups and Rho inhibitation group reduced signifi-cantly([1003.32 ±3.42] ng/L, [821.32 ±21.32] ng/L, [590.15 ±7.68] ng/L, [488.13 ±65.03] ng/L vs [1 347.54 ± 41.52]ng/L,P<0.05), while the expressions of IL-6 mRNA reduced accordingly in 5, 10, 20 mg/mL rHuEPO intervention groups and Rho inhibitation group reduced significantly ([656.68 ±0.55] pg/L, [422.35 ±0.22] pg/L, [217.32 ±0.35] pg/L, [309.49 ±0.21] pg/L vs [884.62 ±0.73]pg/L,P<0.05).Moreover, there was significant statistical difference among 5, 10, 20 mg/mL rHuEPO intervention groups(P<0.05). Conclusion RHuEPO can inhibit the transdifferentiation process of HK-2 cells in-duced by albumin by suppressing inflammation factors , and the mechanism may be involved in RhoA/ROCK signaling pathway .

The effect and significance of rhEPO to the expression of BCL-2 and Caspase-9 in the new type BPD model’ s lung tissue / 安徽医科大学学报

Objective To explore the role of phosphatidylinositol 3-kinase/protein kinase B(PI3K/AKT) pathway in recombinant human erythropoietin( rhEPO) protecting new type BPD. Methods Injected LPS or PBS to 40 spra-gue-dawley (SD) rats, gestational sacs on 15th day of gestation, the newborn rats were divided into four groups:control group, hyperoxia group, rhEPO group, rhEPO+LY294002 group. To detect the expression of lung tissue BCL-2 and Caspase - 9 protein and mRNA by Western blot and RT-PCR. Results In the hyperoxia group and rhEPO+LY294002 group, BCL-2 expression was decreased,while Caspase-9 expression was increased apparently;in the rhEPO group, BCL-2 expression was increased, while Caspase-9 expression was decreased. The differences between each group were statistically significant on 1st,7th and 14th day of hyperoxia exposure (P<0. 05). Con-clusion Intrauterine inflammatory exposure combined with hyperoxia after birth can cause lung cell apoptosis in-creases. While rhEPO has certain control effects on BPD, possibly by reducing the pulmonary apoptosis. The mechanism may be associated with PI3K/AKT signaling pathway.

Recombinant human erythropoietin attenuates pulmonary inflammatory in newborn rats with chronical hyperoxia-induced bronchopulmonary dysplasia / 中华实用儿科临床杂志

Objective To investigate anti-inflammatory effect of recombinant human erythropoietin(rhEPO) on bronchopulmonary dysplasia in newborn rats exposed to hyperoxia.Methods Ninety-six Wistar newborn rats were randomly divided into 4 groups after birth:room air-exposed control group,room air-exposed rhEPO treated group,hyperoxia-exposed group,and the hyperoxia-exposed rhEPO treated group.The last two groups were exposed to oxygen,FiO2 =850 mL/L,room air-exposed rhEPO treated and hyperoxia-exposed rhEPO treated group received rhEPO 2 400 IU/kg subcutaneously at birth,30 minutes' before oxygen exposure and 2 d after birth.The isodose of 9 g/L saline was given in the same way in room air-exposed controls and hyperoxia-exposed pups.Rats from each group were sacrificed on day 3,7 and 10.Lung histology was observed under microscope,and mRNA expression of monocyte chemoattractant protein-1 (MCP-1) and cytokine-induced neutrophil hemoattractant-1 (CINC-1) were determined with reverse transcriotion-polymerase chain reaction(RT-PCR).Results Under microscope,in the hyperoxia-exposed group,inflammatory cell influx was detected in the lungs on the 3rd day and there was marked neutrophlic infiltrate on the 7th day.Alveolar enlargement and fibrosis were evident on the 10th day.At the same time,the histopathological changes were improved greatly in the lungs of hyperoxia-exposed rhEPO treated pups compared with the hyperoxia-exposed pups.MCP-1 and CINC-1 mRNA expression increased in hyperoxia-exposed pups,compared with room air-exposed controls especially on the 7th day [(0.94 ± 0.45) vs (0.21 ± 0.03),P ＜ 0.001 ; (1.26 ± 0.29) vs (0.26 ± 0.06),P ＜ 0.001].MCP-1 and CINC-1 mRNA expression were greatly depressed in the hyperoxia-exposed rhEPO treated pups compared with the hyperoxia-exposed pups especially on the 7th day.[(0.65 ± 0.07) vs (0.94 ± 0.45),P＜0.05;(0.83±0.07) vs (1.26±0.29),P＜0.05].Conclusions The therapy of rhEPO (2 400 IU/kg) therapy can reduce lung inflammatory cell infiltration and alveolar fibrin deposition in newborn rats with hyperoxic lung injury,and it can restrain MCP-1 and CINC-1 mRNA expression.The anti-inflammatory mechanism of rhEPO is related to inhibition of MCP-l and CINC-1 mRNA expression.

Neuroprotective potentials of recombinant human erythropoietin in premature rats with white matter damage / 中华实用儿科临床杂志

Objective To investigate the neuroprotective potentials of recombinant human erythropoietin ( rhEPO) in premature rats with white matter damage. Methods Pregnant rats ( gestational age 15 days) were injected lipopolysaccharide (LPS) (300 μg/kg) intraperitoneally to make cerebral white matter lesions. Another 10 cases of pregnant rats were injected saline (1 mL/kg) intraperitoneally as controls. All preterm rats were born by caesarean sec-tion on embryonic day 21. According to the different processing method,the preterm rats were randomly divided into LPS+rhEPO group (18 cases),LPS+ normal saline (NS) group (18 cases),NS+rhEPO group (18 cases) and NS+NS group (18 cases) ,and they were injected rhEPO (5 000 IU/kg) or 9 g/L saline 1 mL/kg intraperitoneally imme-diately after birth,respectively. The cerebral white matter injury was evaluated with HE staining,and levels of CD68 ,gli-al fibrillary acidic protein (GFAP),myelin basic protein (MBP) were detected by immunofluorescence method 3 and 7 days after birth. Assessment of nerve behavior was done 2 weeks after birth. Results HE staining showed that the white matter lesions were less in LPS+rhEPO group than those in LPS+NS group 3 and 7 days after birth,while NS+rhEPO group and NS+NS group had no cerebral white matter lesions. The expressions of CD68 in LPS+rhEPO group,NS+rhEPO group,NS+NS group significantly decreased compared with LPS+NS group (F=7. 456,P0. 05). No significant differences were found between LPS+rhEPO group and the other groups in evaluation of long-term neural development(all P>0. 05). The val-ues of F by the open field test,suspension test,slope hill test,and resistance to capture test were 2. 09,0. 53,0. 11,0. 37, respectively. Conclusions A single large dose (5 000 IU/kg) rhEPO has neuroprotective effect on the cerebral white matter lesions in the premature rats by inhibiting microglia and astrocyte activation in a short time. The long-term effort remains unknown.

Effect of recombinant murine erythropoietinon injected into mice with multiple myeloma on their immune function / 西安交通大学学报(医学版)

Objective To investigate the effect of recombinant human erythropoietin on immune function in mice with multiple myeloma.Methods Multiple myeloma model mice were divided randomly into control group and treatment groups (low,middle and high doses).The rats in treatment groups were injected with recombinant human erythropoietin 100 μL (2.5,5.0 and 10 mg/kg)via the tail vein for 30 days,and the rats in control group were injected isotonic saline solution of the same volume instead.After the experiment,the colorimetric MTT assay,neutral red method and ELISA kit were used to evaluate the proliferation of B and T lymphocytes,the phagocytic ability of macrophages and the content of serum TNF-α.Results Compared with those in control group,different doses of recombinant human erythropoietin could significantly increase the proliferation of lymphocytes,increase the phagocytosis of macrophages and improve the production of serum TNF-α(P < 0.05 ). Conclusion Recombinant human erythropoietin can significantly improve immunoregulation of mice with multiple myeloma.

The effect of rhEPO on the expression of Bcl-2 and Caspase-3 in intestinal tract of newborn rats with necrotizing enterocolitis / 临床儿科杂志

Objectives To investigate the effect of rhEPO on newborn rats with necrotizing enterocolitis (NEC). Meth-ods Sixty newborn Sprague-Dewley (SD) rats at the age of 48 hours were randomly divided into 5 groups:control group, NEC group, and intervention groups 1, 2 and 3 treated with rhEPO. The rats were fed rat breast milk substitutes and stressed under hypoxia and cold exposure to establish NEC model. The rats with NEC were treated with different doses of rhEPO (0.1U/ml, 1U/ml and 10U/ml) in intervention groups. The expression of Bcl-2 and caspase-3 were measured by immunohistochemistry, and intestinal pathological changes were observed using HE staining. The value of positive expression was analyzed by IOD (integral optical density) image analysis system. Results Abdominal distention, decreased activity and unresponsiveness occurred in NEC rats 24 hours after stress exposure, and pale skin, decreased skin temperature and respiratory rhythm change were observed in severe cases. The symptoms appeared later and milder in three intervention groups. The NEC incidence of newborn rats was as followings:control group(0%), model group(60%), intervention group 1( 30%), intervention group 2(18.2%), intervention group 3(9.1%) and the difference was signiifcant between each group (P=0.008). The grades of intestinal injury, the expression of active caspase-3 and Bcl-2 were signiifcantly different among groups (P<0.01). Intestinal injury was the most severe and the expression level of active caspase-3 was the highest in NEC group. After rhEPO treatment, the intestinal injury and the production of active caspase-3 protein were decreased, and the expression of Bcl-2 was increased. Conclusions Oral rhEPO could decrease the ex-pression of intestinal active caspase 3, and increase the expression of Bcl-2. The protective effect of rhEPO on NEC is dosede-pendent.