Transcriptomic and bioinformatics analysis of the mechanism by which erythropoietin promotes recovery from traumatic brain injury in mice

Recent studies have found that erythropoietin promotes the recovery of neurological function after traumatic brain injury.However,the precise mechanism of action remains unclea r.In this study,we induced moderate traumatic brain injury in mice by intrape ritoneal injection of erythro poietin for 3 consecutive days.RNA sequencing detected a total of 4065 differentially expressed RNAs,including 1059 mRNAs,92 microRNAs,799 long non-coding RNAs,and 2115circular RNAs.Kyoto Encyclopedia of Genes and Genomes and Gene Ontology analyses revealed that the coding and non-coding RNAs that were differentially expressed after traumatic brain injury and treatment with erythropoietin play roles in the axon guidance pathway,Wnt pathway,and MAPK pathway.Constructing competing endogenous RNA networks showed that regulatory relationship between the differentially expressed non-coding RNAs and mRNAs.Because the axon guidance pathway was repeatedly enriched,the expression of Wnt5a and Ephb6,key factors in the axonal guidance pathway,was assessed.Ephb6 expression decreased and Wnt5a expression increased after traumatic brain injury,and these effects were reversed by treatment with erythro poietin.These findings suggest that erythro poietin can promote recove ry of nerve function after traumatic brain injury through the axon guidance pathway.

Neurotherapeutic potential of erythropoietin after ischemic injury of the central nervous system

Erythropoietin (EPO) is one of the most successful biopharmaceuticals in history and is used for treating anemia of different origins. However, it became clear that EPO could also work in a neuroprotective, antiapoptotic, antioxidative, angiogenetic and neurotropic way. It causes stimulation of cells to delay cell apoptosis, especially in the central nervous system. In rodent models of focal cerebral ischemia, EPO showed an impressive reduction of infarct size by 30% and improvement of neurobehavioral outcome by nearly 40%. A large animal model dealing with ischemia and reperfusion of the spinal cord showed that EPO could reduce the risk of spinal cord injury significantly. In addition, some clinical studies tested whether EPO works in real live clinical settings. One of the most promising studies showed the innocuousness and improvements in follow-up, outcome scales and in infarct size, of EPO-use in humans suffering from ischemic stroke. Another study ended unfortunately in a negative outcome and an increased overall death rate in the EPO group. The most possible reason was the involvement of patients undergoing simultaneously systemic thrombolysis with recombinant tissue plasminogen activator. An experimental study on rats demonstrated that administration of EPO might exacerbate tissue plasminogen activator-induced brain hemorrhage without reducing the ischemic brain damage. This case shows clearly how useful animal models can be to check negative side effects of a treatment before going into clinical trials. Other groups looked in human trials at the effects of EPO on the outcome after ischemic stroke, relation to circulating endothelial progerdtor cells, aneurysmal subarachnoid hemorrhage, traumatic brain injury, hemoglobin transfusion thresholds and elective first-time coronary artery bypass surgery. Most of the results were pos-让ive, but are based mostly on small group sizes. However, some of the most neglected facts when focusing on experimental setups of ischemia of the central nervous system are issues like age and comorbidities. It might be extremely worthy to consider these points for future projects, because EPO might influence all these factors.

Does recombinant human erythropoietin accelerate correction of post-ulcer-bleeding anaemia?A pilot study

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 after the endoscopic confirmation of cessation of bleeding,they were randomised either to erythropoietin(20 000 IU Epoetin alfa subcutaneously,on days 0,4 and 6)plus iron(100 mg im,on days 1-6,(G_1)or iron only(G_2).Haematocdt was measured on days 0,6,14, 30,45,and 60,respectively. RESULTS:One patient from G_1 and two from G_2 were lost to follow-up.Therefore,14 and 13 patients from G_1 and G_2 respectively were analysed.Demographic characteristics,serum iron,ferritin,total iron binding capacity,reticulocytes,and haernatoait were not significantly different at entry to the study. Median reticulocyte counts were significantly different between groups on day six(G_1:4.0,3.0-6.4 vs G_2:3.5,2.1-4.4%, P=0.03)and median haematocrit on day fourteen [G_1:35.9, 30.7-41.0 vs G_2:32.5,29.5-37.0%(median,range),P=0.04]. CONCLUSION:Erythropoietin administration significantly accelerates correction of anemia after acute ulcer bleeding. The haematocrit gain is equivalent to one unit of transfused blood two weeks after the bleeding episode.

Constitutively Active Soluble Form of Erythropoietin Receptor Suppresses Growth and Angiogenesis of Xenografts of Transfected Cancer Cell Lines

We examined the effect of blocking the erythropoietin (Epo) signaling using an anti-Epo antibody, soluble form of Epo receptor (sEpoR) capable of binding to Epo or EpoR antagonist, and proved to be effective against xenografts of female reproductive organ malignancies and of cancer cell lines in nude mice. We transfected seven cancer cell lines of various origins to express constitutively active sEpoR, and examined their tumorigenesis in nude mice. Suppression of the tumor growth, decrease in viable and proliferating cells and reduction of vascular density were seen individually in all xenografts of transfected cell lines compared with the controls. Quantitative RT-PCR analyses showed that expression levels of Epo, EpoR, ?1A-adrenaline receptor (?1A-ADR) and muscalinic acetylcholine receptor subunit 3 mRNAs (m3-AchR) were higher in the majority of the wild-type xenografts than in the corresponding cell lines except for A549. In some of the transfected xenografts, EpoR, ?1A-ADR and m3-AchR mRNAs were down-regulated. Western blot analyses revealed that the constitutively activated ERK1/2MAPK was discernible in the majority of non-transfected cell lines and was reduced in the transfected cell lines. However, it was regained after exposure to acetylcholine and/or noradrenaline. These findings suggest that constitutively active sEpoR can effectively destroy the xenografts but signals from the autonomic neurotransmitters of the host produced under stress may interfere with this antitumor activity.

Evaluation of Hemoglobin and Serum Erythropoietin Levels in Patients with Polycythemia Vera and Secondary Polycythemia

Objective: This study compares hemoglobin and erythropoietin levels in patients with polycythemia vera and secondary polycythemia. Study Design: A retrospective cross-sectional study evaluating the serum erythropoietin and hemoglobin levels in patients with polycythemia vera and secondary polycythemia. This study was performed simultaneously in Texas state of the U.S. and Fars Province in Iran. Methods: Hemoglobin, hematocrit and erythropoietin test results were collected from patients aged 19 to 75 years who were diagnosed with polycythemia vera and secondary polycythemia. Patients records with history of thrombocythemia, congestive heart failure, dyspnea, anemia and pregnant woman were excluded from study. Patients in each decade of life were examined in separate groups, so that changes in hemoglobin related to aging did not affect the research results. Results: 75% of the patients were men, and 25% were women. A total of 1580 patients were analyzed in this study. 57.3% of patients in UTMB and 38.8 patients in Iran have hemoglobin level above 17 mg/dl. 74% of patients in UTMB and 88% of patients in Iran have erythropoietin below 10 IU/mL. Polycythemia in UTMB was more common in people over 50 and in Iran in patients under 50 years old. The serum hemoglobin and erythropoietin levels in patients with polycythemia vera were not significantly different in compare to secondary polycythemia patients. Data showed that there were 84 polycythemia patients per 100,000 people. The results of this study in UTMB and Iran showed that 4.5% and 7%, respectively, of patients with polycythemia had a positive JAK2 test. Conclusion: Low erythropoietin levels may not be helpful in differentiating polycythemia vera from secondary polycythemia. .

Plasma erythropoietin levels in anaemic and non-anaemic patients with chronic liver diseases

AIM: To investigate the serum erythropoietin (Epo) levels in patients with chronic liver diseases and to compare to subjects with iron-deficiency anaemia and healthy controls.METHODS: We examined 31 anaemic (ALC) and 22 non-anaemic (NALC) cirrhotic patients, 21 non-anaemic subjects with chronic active hepatitis (CAH), 24 patients with iron-deficiency anaemia (ID) and 15 healthy controls. Circulating Epo levels (ELISA; R&D Systems, Europe Ltd, Abingdon,UK) and haemoglobin (Hb) concentration were determined in all subjects.RESULTS: Mean±SD of Epo values was 26.9±10.8 mU/mL in ALC patients, 12.5±8.0 mU/mL in NALC subjects,11.6±6.3 mU/mL in CAH patients, 56.4±12.7 mU/mL in the cases of ID and 9.3±2.6 mU/mL in controls. No significant difference (P>0.05) was found in Epo levels between controls, CAH and NALC patients. ALC individuals had higher Epo levels (P<0.01) than these groups whereas ID subjects had even higher levels (P<0.001) than patients suffering from ALC.CONCLUSION: Increased Epo values in cirrhotics, are only detectable when haemoglobin was lesser than 12 g/dL.Nevertheless, this rise in value is lower than that observed in anaemic patients with iron-deficiency and appears blunted and inadequate in comparison to the degree of anaemia.

Effect of erythropoietin combined with PCI on myocardial damage and vascular endothelial function in patients with STEMI

Objective:To investigate the effects of erythropoietin combined with percutaneous coronary intervention (PCI) on myocardial damage, vascular endothelial function and neuroendocrine hormone levels in patients with ST-segment elevation myocardial infarction (STEMI).Method:From August 2015 to February 2018, 80 patients with STEMI were selected from our hospital, divided into 2 groups according to admission time, 40 cases in each group, set to observation group and control group, the observation group was treated with erythropoietin combined with PCI, and the control group was treated only with PCI, to compare the degree of myocardial damage, vascular endothelial function and neuroendocrine hormone levels after treatment. Results: At 24 h after surgery, the observed group of cardiac troponin (cTNI), creatine kinase isoenzyme (CK-MB), type I collagen carboxy terminal peptide (ICTP) and ischemic modified albumin (IMA) is lower than the control group,the cystatin-C (Cys-C) level is higher than the control group, the difference was statistically significant (P<0.05);At 1 week after surgery, the levels of endothelin-1 (ET-1) and von Willebrand factor (vWF) in the observation group were lower than the control group, and the level of nitric oxide (NO) was higher than that in the control group, the difference was statistically significant (P<0.05);At 1 week after surgery, the levels of cortisol (Cor), adrenocortical hormone (ACTH), norepinephrine (NE) and Angiotensin II (AngII) in the observation group were lower than in the control group, the difference was statistically significant (P<0.05).Conclusion: Erythropoietin combined with PCI in the treatment of STEMI patients with better clinical effect, can effectively reduce the degree of myocardial damage, protect vascular endothelial cell function, improve the body's neuroendocrine hormone levels, recommend clinical application.

Outbreak of methanol-induced optic neuropathy in early COVID-19 era;effectiveness of erythropoietin and methylprednisolone therapy

BACKGROUND Methanol is a highly toxic,non-potable alcohol.Outbreaks of methanol toxicity occur due to its fraudulent addition to alcoholic beverages as a cheaper substitute for ethanol.Recently,alongside the coronavirus disease 2019(COVID-19)pandemic,rumors circulated on social media that consuming alcohol can prevent or cure the virus,leading to a COVID-19 and methanol-induced optic neuropathy(MON)syndemic.AIM To investigate the impact of erythropoietin(EPO)on the outcomes of patients diagnosed with MON.METHODS In this prospective study,105 patients presenting with acute bilateral visual loss secondary to methanol intoxication were enrolled from March to May 2020 at Farabi Eye Hospital.A comprehensive ocular examination was conducted for all participants.Recombinant human EPO and methylprednisolone were administered intravenously to all patients for three consecutive days.RESULTS The mean age of the participants was 39.9 years(±12.6).Ninety-four patients were male and eleven were female.The mean pre-treatment best corrected visual acuity(BCVA)improved from 2.0±0.86 to 1.39±0.69 logarithm of the minimum angle of resolution post-treatment(P<0.001),with significant improvement observed in all age categories and genders(P<0.001).Visual acuity improvement was also significant regardless of whether the patient presented before or after 72 h(P<0.001),and the post-treatment BCVA remained significant at all monthly follow-up visits(P<0.001).CONCLUSION EPO and methylprednisolone therapy have been shown to be effective in improving visual outcomes in patients with MON when administrated within the first month of exposure.Public awareness efforts are necessary to prevent further outbreaks of methanol toxicity in the current COVID-19 era.

Transplantation of erythropoietin gene-modified neural stem cells improves the repair of injured spinal cord

The protective effects of erythropoietin on spinal Here, the eukaryotic expression plasmid pcDNA3.1 cord injury have not been well described. human erythropoietin was transfected into rat neural stem cells cultured in vitro. A rat model of spinal cord injury was established using a free falling object. In the human erythropoietin-neural stem cells group, transfected neural stem cells were injected into the rat subarachnoid cavity, while the neural stem cells group was inject- ed with non-transfected neural stem cells. Dulbecco＇s modified Eagle＇s medium/F12 medium was injected into the rats in the spinal cord injury group as a control. At 1-4 weeks post injury, the motor function in the rat lower limbs was best in the human erythropoietin-neural stem ceils group, followed by the neural stem cells group, and lastly the spinal cord injury group. At 72 hours, compared with the spinal cord injury group, the apoptotic index and Caspase-3 gene and protein expressions were apparently decreased, and the bd-2 gene and protein expressions were noticeably increased, in the tissues surrounding the injured region in the human erythro- poietin-neural stem cells group. At 4 weeks, the somatosensory evoked potential latencies were cavities were clearly smaller and the motor and remarkably shorter in the human erythropoi- etin-neural stem cells group and neural stem cells group than those in the spinal cord injury group. These differences were particularly obvious in the human erythropoietin-neural stem cells group. More CM-Dil-positive cells and horseradish peroxidase-positive nerve fibers and larger amplitude motor and somatosensory evoked potentials were found in the human erythro- poietin-neural stem cells group and neural stem cells group than in the spinal cord injury group. Again, these differences were particularly obvious in the human erythropoietin-neural stem cells group. These data indicate that transplantation of erythropoietin gene-modified neural stem cells into the subarachnoid cavity to help repair spinal cord injury and promote the recovery of spinal cord function better than neural stem cell transplantation alone. These findings may lead to significant improvements in the clinical treatment of spinal cord injuries.

Impact of erythropoietin therapy on cardiorenal syndrome:A systematic review with meta-analysis

BACKGROUND Heart and kidney dysfunction frequently coexist in patients with acute heart failure due to the overlap between these two organ systems.Cardiorenal syndrome(CRS)results from pathology occurring in the heart and kidneys along with the consequences of dysfunction in one organ contributing to dysfunction in the other and vice versa.AIM To evaluate the use of erythropoietin(EPO)in patients with CRS and its effects on hemoglobin(Hb),major cardiovascular(CV)events,and hospitalization rates.METHODS On February 24,2022,searches were conducted using PubMed,MEDLINE,and EMBASE,and 148 articles were identified.A total of nine studies were considered in this systematic review.We assessed the included articles based on the National Heart,Lung,and Blood Institute quality assessment tools for controlled intervention and observational cohort or cross-sectional studies.An assessment of bias risk was conducted on the chosen studies,and data relevant to our review was extracted.RESULTS The systematic review of these studies concluded that most existing literature indicates that EPO improves baseline Hb levels and decreases myocardial remodeling and left ventricular dysfunction without reducing CV mortality.In addition,the effect of EPO on the hospitalization rate of patients with CRS needs to be further studied since this relationship is unknown.Future studies,such as randomized controlled clinical trials and prospective cohort studies,should be conducted to enhance the literature on the potential of EPO therapy in patients with CRS.CONCLUSION Our systematic review suggests that EPO therapy may have a significant role in managing CRS.The review highlights the potential benefits of EPO in improving baseline Hb levels,reducing the risk of major CV events,improving cardiac remodeling,myocardial function,New York Heart Association class,and B-type natriuretic peptide levels.However,the effect of EPO treatment on hospitalization remains unclear and needs further exploration.

Improved Recovery of Erythropoietin and Darbepoetin from Equine Plasma by the Application of a Wheat Germ Agglutinin Mediated Pre-Extraction Prior to Immunoaffinity Chromatography

We describe a two-step method that uses wheat germ agglutinin immobilized on Sepharose gel followed by immunoaffinity chromatography (IAC) to extract recombinant human erythropoietin and Darbepoetin from equine plasma. Lectin affinity chromatography was shown to be an effective approach for isolating these epoetins from plasma and in combination with IAC;this method gave superior recovery when compared to the use of the latter technique alone. Moreover, due to the ease with which it can be scaled up, it is particularly well suited for pre-concentrating larger volumes of samples prior to IAC and this provides a facile way of improving the overall sensitivity with which these foreign proteins can be detected in equine plasma.

Novel nervous and multi-system regenerative therapeutic strategies for diabetes mellitus with mTOR

Throughout the globe,diabetes mellitus（DM） is increasing in incidence with limited therapies presently available to prevent or resolve the significant complications of this disorder.DM impacts multiple organs and affects all components of the central and peripheral nervous systems that can range from dementia to diabetic neuropathy.The mechanistic target of rapamycin（m TOR） is a promising agent for the development of novel regenerative strategies for the treatment of DM.m TOR and its related signaling pathways impact multiple metabolic parameters that include cellular metabolic homeostasis,insulin resistance,insulin secretion,stem cell proliferation and differentiation,pancreatic β-cell function,and programmed cell death with apoptosis and autophagy.m TOR is central element for the protein complexes m TOR Complex 1（m TORC1） and m TOR Complex 2（m TORC2） and is a critical component for a number of signaling pathways that involve phosphoinositide 3-kinase（PI 3-K）,protein kinase B（Akt）,AMP activated protein kinase（AMPK）,silent mating type information regulation 2 homolog 1（Saccharomyces cerevisiae）（SIRT1）,Wnt1 inducible signaling pathway protein 1（WISP1）,and growth factors.As a result,m TOR represents an exciting target to offer new clinical avenues for the treatment of DM and the complications of this disease.Future studies directed to elucidate the delicate balance m TOR holds over cellular metabolism and the impact of its broad signaling pathways should foster the translation of these targets into effective clinical regimens for DM.

Effect of Recombinant Human Erythropoietin on Survivin Expression in Brain Tissues after Traumatic Brain Injury in Rats

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.

Cardioprotective effect of erythropoietin on sepsisinduced myocardial injury in rats

BACKGROUND:Sepsis-induced myocardial injury is one of the major predictors of morbidity and mortality of sepsis.The cytoprotective function of erythropoietin(EPO) has been discovered and extensively studied.However,the cardioprotective effects of EPO on sepsis-induced myocardial injury in the rat sepsis model has not been reported.METHODS:The rat models of sepsis were produced by cecal ligation and perforation(CLP)surgery.Rats were randomly(random number) assigned to one of three groups(n=8 for each group):sham group,CLP group and EPO group(1000 lU/kg erythropoietin).Arterial blood was withdrawn at3,6,12,and 24 hours after CLP.cTnl,BNP,CK-MB,LDH,AST,TNF-a,IL-6,IL-10,and CRP were tested by the ELISA assay.Changes of hemodynamic parameters were recorded at 3,6,12,24 hours after the surgery.Histological diagnosis was made by hematoxylin and eosin.Flow cytometry was performed to examine cell apoptosis,myocardium mitochondrial inner membrane potential,and NF-κB(p65).Survival rate at 7 days after CLP was recorded.RESULTS:In the CLP group,myocardial enzyme index and inflammatory index increased at3,6,12 and 24 hours after CLP compared with the sham group,and EPO significantly blocked the increase.Compared with the CLP group,EPO significantly improved LVSP,LV +dpldt_(max) LV-dp/dt_(min),and decreased LVEDP at different time.EPO blocked the reduction of mitochondrial transmembrane potential,suppressed the cardiomyocyte apoptosis,inhibited the activation of NF-κB,and reduced the production of proinflmmatory cytokines.No difference in the survival rate at 7 days was observed between the CLP group and the EPO group.CONCLUSION:Exogenous EPO has cardioprotective effects on sepsis-induced myocardial injury.

Erythropoietin inhibits ferroptosis and ameliorates neurological function after spinal cord injury

Ferroptosis is one of the critical pathological events in spinal cord injury.Erythropoietin has been reported to improve the recovery of spinal cord injury.However,whether ferroptosis is involved in the neuroprotective effects of erythropoietin on spinal cord injury has not been examined.In this study,we established rat models of spinal cord injury by modified Allen’s method and intraperitoneally administered 1000 and 5000 IU/kg erythropoietin once a week for 2 successive weeks.Both low and high doses of erythropoietin promoted recovery of hindlimb function,and the high dose of erythropoietin led to better outcome.High dose of erythropoietin exhibited a stronger suppressive effect on ferroptosis relative to the low dose of erythropoietin.The effects of erythropoietin on inhibiting ferroptosis-related protein expression and restoring mitochondrial morphology were similar to those of Fer-1(a ferroptosis suppressor),and the effects of erythropoietin were largely diminished by RSL3(ferroptosis activator).In vitro experiments showed that erythropoietin inhibited RSL3-induced ferroptosis in PC12 cells and increased the expression of xCT and Gpx4.This suggests that xCT and Gpx4 are involved in the neuroprotective effects of erythropoietin on spinal cord injury.Our findings reveal the underlying anti-ferroptosis role of erythropoietin and provide a potential therapeutic strategy for treating spinal cord injury.

Effect of carbamylated erythropoietin on retinopathy of diabetic rats

Objective:To study the effect of carbamylated erythropoietin (CEPO) on retinopathy of diabetic rats.Methods: Male SD rats were selected as experimental animals and randomly divided into control group, DM group and CEPO group, and diabetic animal models were established and then given CEPO intervention. 2 weeks after intervention, the retina was collected to detect the expression of angiogenesis molecules, apoptosis molecules and oxidative stress pathway molecules.Results: HIF-1α, VEGF, Ang-1, Bax, Caspase-3, Nrf-2, ARE, HO-1 and NQO-1 mRNA expression in retina of DM group were significantly higher than those of control group while TKLK, PEDF, Bcl-2 and Survivin mRNA expression were significantly lower than those of control group;HIF-1α, VEGF, Ang-1, TKLK and PEDF mRNA expression in retina of CEPO group were not significantly different from those of DM group, Bcl-2, Survivin, Nrf-2, ARE, HO-1 and NQO-1 mRNA expression were significantly higher than those of DM group, and Bax and Caspase-3 mRNA expression were significantly lower than those of DM group.Conclusion:CEPO can reduce the apoptosis and oxidative stress injury of the retina tissue in diabetic rats without affecting the angiogenesis.

Effect of recombinant human erythropoietin therapy on convalescent serological indicators in patients with severe craniocerebral injury

Objective:To study the effect of recombinant human erythropoietin (rHu-EPO) therapy on convalescent serological indicators in patients with severe craniocerebral injury.Methods:Patients with severe craniocerebral injury who were treated in Fifth Hospital in Wuhan between July 2014 and February 2017 were selected and randomly divided into the rHu-EPO group who accepted rHu-EPO combined with conventional therapy and the control group who accepted conventional therapy. Before and after treatment, serum levels of nerve injury indexes, inflammation indexes, oxidative stress indexes and apoptosis indexes were measured. Results:Serum Tau, S100B, GFAP, NSE, IL-1β, TNF-α, VCAM-1, ICAM-1, LPO, AOPP, 8-iso-PGF2 , sTRAIL, sFas and sFasL levels of both groups of patients 14 d after treatment were significantly lower than those before treatment, and serum Tau, S100B, GFAP, NSE, IL-1β, TNF-α, VCAM-1, ICAM-1, LPO, AOPP, 8-iso-PGF2 , sTRAIL, sFas and sFasL levels of rHu-EPO group were significantly lower than those of control group.Conclusion: rHu-EPO therapy can significantly improve the convalescent nerve injury, inflammation, oxidative stress and apoptosis in patients with severe craniocerebral injury.

Age and genotype dependent erythropoietin protection in COVID-19

Erythropoietin(EPO)is the main mediator of erythropoiesis and an important tissue protective hormone that appears to mediate an ancestral neuroprotective innate immune response mechanism at an early age.When the young brain is threatened-prematurity,neonatal hyperbilirubinemia,malaria-EPO is hypersecreted disproportionately to any concurrent anemic stimuli.Under eons of severe malarial selection pressure,neuroprotective EPO augmenting genetic determinants such as the various hemoglobinopathies,and the angiotensin converting enzyme(ACE)I/D polymorphism,have been positively selected.When malarial and other cerebral threats abate and the young child survives to adulthood,EPO subsides.Sustained high ACE and angiotensin II(Ang II)levels through the ACE D allele in adulthood may then become detrimental as witnessed by epidemiological studies.The ubiquitous renin angiotensin system(RAS)influences theα-klotho/fibroblast growth factor 23(FGF23)circuitry,and both are interconnected with EPO.Here we propose that at a young age,EPO augmenting genetic determinants through ACE D allele elevated Ang II levels in some or HbE/beta thalassemia in others would increase EPO levels and shield against coronavirus disease 2019,akin to protection from malaria and dengue fever.Human evolution may use ACE2 as a“bait”for severe acute respiratory syndrome coronavirus-2(SARS-CoV-2)to gain cellular entry in order to trigger an ACE/ACE2 imbalance and stimulate EPO hypersecretion using tissue RAS,uncoupled from hemoglobin levels.In subjects without EPO augmenting genetic determinants at any age,ACE2 binding and internalization upon SARS-CoV-2 entry would trigger an ACE/ACE2 imbalance,and Ang II oversecretion leading to protective EPO stimulation.In children,low nasal ACE2 Levels would beneficially augment this imbalance,especially for those without protective genetic determinants.On the other hand,in predisposed adults with the ACE D allele,ACE/ACE2 imbalance,may lead to uncontrolled RAS overactivity and an Ang II induced proinflammatory state and immune dysregulation,with interleukin 6(IL-6),plasminogen activator inhibitor,and FGF23 elevations.IL-6 induced EPO suppression,aggravated through co-morbidities such as hypertension,diabetes,obesity,and RAS pharmacological interventions may potentially lead to acute respiratory distress syndrome,cytokine storm and/or autoimmunity.HbE/beta thalassemia carriers would enjoy protection at any age as their EPO stimulation is uncoupled from the RAS system.The timely use of rhEPO,EPO analogs,acetylsalicylic acid,bioactive lipids,or FGF23 antagonists in genetically predisposed individuals may counteract those detrimental effects.

Cell-Based Biological Markers for Blood-Enriching Chinese Herbs: Erythropoietin Production in HepG2 Cells and Nitric Oxide Release in Human Umbilical Vein Endothelial Cells (HUVECs)

Chinese tonifying herbs, which are classified into four functional categories (namely, Yang, Qi, Yin, Blood) are commonly used for restoring normal body function and the prevention of diseases. To explore cell-based biological markers for Blood-enriching Chinese herbs, we investigate the effect of 11 commonly used Blood-enriching herbs on erythropoietin (EPO) production in HepG2 cells. Herbs for nourishing Yin were tested for determining the specificity of Blood-enriching herbs in inducing EPO production. In addition, the effects of Blood-enriching herbs on nitric oxide (NO) production in both HepG2 cells and human umbilical vein endothelial cells (HUVECs) were also investigated. The results indicated that methanolic extracts of Blood-enriching herbs (but not Yin-nourishing herbs) showed characteristic pharmacological activity in inducing EPO production in HepG2 cells and NO release in HUVECs. The experimental findings, therefore, support the use of cell-based EPO production and NO release as biological markers for Blood-enriching Chinese tonifying herbs.

Assessment of Patient Perceptions about Use of Wepox Pen^TM(Recombinant Erythropoietin Delivery Device with 30,000 IU Cartridge) in the Management of Anemia in Chronic Kidney Disease Patients

Objective: To assess perceptions about ease of use and other benefits of Wepox PenTM (loaded with 30,000 IU cartridge of recombinant erythropoietin) in the management of anemia in adult chronic kidney disease (CKD) patients. Material and methods: In this prospective, observational, multicentric post marketing surveillance, adult CKD patients treated with erythropoietin were enrolled from November 2015 to December 2016 to understand their opinions about Wepox PenTM. Ease of use of pen, ease of administering accurate dose, confidence in administration and ease of storage and disposal of cartridge were assessed on five points Likert scale: 1. very easy;2. somewhat easy;3. neither easy nor difficult;4. somewhat difficult;5. very difficult. Global assessment was performed on five points scale: 1. excellent;2. very good;3. good;4. average;5. not good. Safety was recorded by checking pain and discomfort and adverse events. Results: A total of 263 patients (mean age 32.87 years;66% male;34% female) were enrolled. Number of patients reporting ease of use as “very easy” from 209 (80.7%) at baseline increased to 245 (94.6%) and 249 (96.1%) at visit 2 (p = 0.001) and visit 3 (p = 0.001) respectively. Number of “very easy” response for accurate dose increased from 236 (91.1%) at visit 1 to 246 (95%) at visit 2 (p = 0.84) and 249 (96.1%) at visit 3 (p = 0.001). Number of the patients with “no pain” at injection site increased from 177 (68.3%) at visit 1 to 205 (79.2%) and 212 (81.9%) at visit 2 (p = 0.001) and visit 3 (p = 0.001) respectively. Improvement in number of patients with “no hurt” at visit 2 (p = 0.538) and visit 3 (p = 0.286) was not statistically significant. Number of patients reporting “somewhat easy” to “very easy” confidence in self injection increased from 251 (96.9%) at visit 2 to 255 (98.5%) at visit 3. Number of patients reporting ease of storage and disposal of cartridge as “somewhat easy” to “very easy” increased from 254 (98.1%) at visit 2 to 256 (98.9%) at visit 3. According to the global assessment, 144 (56.3%) cases reported “excellent” response. “Very good” and “Good” responses were reported by 106 (41.4%) and 6 (2.3%) patients respectively. A total of 230 (98.7%) patients said that they would prefer to use erythropoietin pen device for further treatment too. Conclusion: Wepox PenTM(recombinant erythropoietin) is easy to use and does not cause significant pain or discomfort. Ability to self-administer recombinant erythropoietin with Wepox PenTM is a great advantage which can make a significant difference for both CKD patients and doctors. Storage and disposal of cartridge is also easy.