Prostate cancer occurs frequently in men and can often lead to death. Many cancers, including prostate cancer, can be initiated by oxidative insult caused by free radicals and reactive oxygen species. The superoxide dismutase family removes the oxygen-derived reactive oxygen species, and increased superoxide dismutase activity can often be protective against prostate cancer. Prostate cancer can be treated in a variety of ways, including surgery, androgen deprivation therapy, radiation therapy, and chemotherapy. The clinical trajectory of prostate cancer varies from patient to patient, but more aggressive tumors often tend to be radioresistant. This is often due to the free-radical and reactive-oxygen-species-neutralizing effects of the superoxide dismutase family. Superoxide dismutase 2, which is especially important in this regard, can be induced by the NF-κB pathway, which is an important mechanism in radioresistance. This information has enabled the development of interventions that manipulate the NF-κB mechanism to treat prostate cancer. In 2021, almost 250,000 men in the United States are projected to be diagnosed with prostate cancer and almost 34,000 are projected to die from the disease [Treatments for prostate cancer include surgery, radiation, and androgen deprivation, among other therapies. Often these therapies are used in combination with each other. Radiotherapy kills cancer cells by generating free radicals and reactive oxygen species (ROS) that progress to cause fatal cellular and DNA damage. Unfortunately, in some patients, the disease, including castration-resistant prostate cancer and radioresistant prostate cancer, resists therapy.Mutations in cellular DNA often occur due to insults caused by free radicals and ROS, which can be generated by cellular processes or exogenous causes such as radiation. The superoxide dismutase (SOD) family consists of SOD1, SOD2, and SOD3 proteins that neutralize the first oxygen-derived ROS and provide protection against the development of cancers, including prostate cancer.Even though SOD can have antitumor effects, it can also contribute to radioresistance, which is often mediated by the NF-κB pathway. One effect of the NF-κB pathway is that transcription of the SOD2 gene increases. In that case, a side effect of neutralizing the ROS is that the cancer cells survive radiotherapy and have prometastasic characteristics.In this minireview, we review first the basics of prostate cancer and its treatment; then, the role of the NF-κB pathway in SOD induction and radioresistance; next, the role of the SOD family in prostate tissue and prostate cancer; and finally focus on the present translational interventions that utilize the NF-κB pathway and knowledge about SOD.The treatment of prostate cancer is dependent on many factors, including patient performance status, prostate cancer stage, and Gleason score, among others. Treatments include surgery, androgen-deprivation therapy, and radiation therapy for most nonmetastatic cases. Chemotherapy is usually used only for metastatic cases.Multiple mechanisms are utilized to reduce androgens. Some of the more common options include bilateral orchiectomy, luteinizing hormone-releasing hormone (LHRH) agonists, and antiandrogens. A list of androgen-lowering pharmacologic agents can be found in Radiation therapy, encompassing brachytherapy and external beam radiation, is a commonly utilized curative modality for the treatment of localized prostate cancer, with outcomes comparable to radical prostatectomy . In addiThe generation of ROS is critical for the efficacy of radiation therapy, but it can activate signaling pathways that contribute to radiation resistance and tumor recovery . There aIt is important to note that antioxidant enzymes often do not work alone in the prevention of oxidative damage, but often work in tandem. For example, SOD converts two superoxide radicals (and two hydrogen ions) into hydrogen peroxide and molecular oxygen. Then, other enzymes, such as catalase or peroxidase, further convert two molecules of hydrogen peroxide into water and oxygen. There are several antioxidant enzymes in addition to SOD and catalase. For example, peroxiredoxins (Prx) are an interesting and relatively newly discovered group of enzymes that scavenge peroxide and peroxynitrite. Glutathione, a cellular thiol found in both the cytoplasm and organelles, also reduces ROS and free radicals.All the antioxidants have been implicated in radioresistance and are topics of active research in varying degrees of emphasis. Peroxiredoxins have a complicated relationship with radioresistance. Prx 2 is significantly upregulated in radioresistant tumors, but Prx 1, 3, 4, 5, and 6 are not . A studyImportantly, molecular pathways governing the transition to castration resistance are often implicated in radiation therapy failure as well . Redox-sExposure to increased oxidative stress is one of the factors which may be implicated in the transformation to more aggressive and castration-resistant prostate cancer . The resThe NF-κB family of proteins includes members p105/p50 (NFκB1), p100/p52 (NFκB2), c-Rel, RelB, and RelA, which are transcription factors that play a critical role in governing cellular processes including growth, survival, and apoptosis . The NF-NF-κB is capable of activating over 150 target genes, including inflammatory cytokines, growth factors, and cell adhesion molecules . This acIncreased NF-κB nuclear activity has been found to be associated with progression of prostate cancer in mouse models . In manyFurthermore, constitutively elevated nuclear levels of RelB are seen in prostate tumors with higher Gleason scores, and correlate with levels of IL-8, a known target gene of the NF-κB pathway and mediator of angiogenesis, which also has other established roles in tumor progression . IL-8 leBecause the NF-κB pathway heavily influences radiosensitivity and radioresistance, certain interventions such as gene therapy or pharmacotherapy can alter the pathway and thus make prostate cancer more radiosensitive. It appears that in androgen-dependent prostate cancer, accumulation of p52 was heavily influenced by activation of the androgen receptor. In a study by Lessard et al., when LNCaP cells were exposed to an androgen analog, more nuclear accumulation of p52 occurred. However, when the antiandrogen bicalutamide was used, nuclear p52 level dropped . A studyImatinib is a tyrosine kinase inhibitor which is specific for abl, c-kit, and the platelet-derived growth factor receptor. It is typically used to treat leukemia. In response to radiotherapy, RelB moves to the nucleus of the cell where it transcribes genes that will lead to radioresistance. Imatinib decreased this nuclear translocation in androgen-independent PC-3 cells and thus increased radiosensitivity. However, imatinib increased RelB nuclear translocation in androgen-sensitive LNCaP cells .2O2 [Free radicals and ROS are generated from mitochondria and other cellular processes, as well as from external sources such as radiation and many pharmacologic agents ,46. A co2O2 . Thus, oOxidative stress and free radical formation have been linked to the development of several malignancies, including prostate cancer. Antioxidant enzymes, such as those in the SOD family, work to protect the cell from oxidative stress, including damage from free radicals and ROS in intra- and extra-cellular environments. The SOD family contains three isoforms in humans, which include SOD1, SOD2, and SOD3. SOD1 is also known as copper–zinc SOD (CuZnSOD) and is located in the cytosol and mitochondrial intermembrane space. SOD2 is also known as manganese SOD (MnSOD) and is present in the mitochondrial matrix. SOD3 is also known as extracellular SOD (EcSOD). It is only present in some cell types and is the only member of the SOD family present in the extracellular matrix, where it protects the cell from extracellular ROS .A study by Botswick et al. examined PC3 cells, prostatic intraepithelial neoplasia (PIN) tissue, and benign epithelium for SOD1, SOD2, and SOD3 using immunohistochemistry, immunogold electron microscopy, and enzymatic assays . The preSome data support that SOD2 levels are increased in intermediate Gleason scores. A study by Miar et al. looked at levels of SOD2 in prostate, colon, and lung cancers, and found increased levels of either the SOD2 protein, mRNA, or both, for all three cancer types. For prostate cancer, the SOD2 protein, but not the mRNA, increased. Though SOD2 expression was increased in all Gleason scores, this was especially true for Gleason scores 6 and 7 . However2O2, and an increase in the proportion of cells in the G1 phase [2O2 can work to reduce proliferation.Interestingly, a study by Venkataraman et al. found that SOD2 expression was reduced in PC-3 cells compared to nonmalignant prostate tissues. When the PC-3 cells were modified to overexpress SOD2, there was a reduction in cell growth rate, an increase in accumulation of extracellular HG1 phase . The accG1 phase ,59. Thus2O2. There is evidence that cells treated with H2O2 have higher levels of 8-OHdG expression [2O2 mediated by overexpression of SOD3 could increase 8-OHdG by causing oxidative DNA damage. This could, in turn, result in tumor cell apoptosis. A paper by Chaiswing et al. points out a correlation between SOD3 overexpression and reduced matrix-metalloproteinase activity, and a corresponding reduced tumor invasion in PC-3 cells [Similar results regarding the overexpression of SOD3 were reported by Kim et al. . Inhibitpression . Thus, i-3 cells .SOD3 function can exert an effect opposite to that of the cysteine/glutamate transporter in the control of the redox state of the tumor microenvironment. Prostate cancer with a high Gleason score has a higher cysteine/glutamate transport expression and lower SOD3 expression. This is similar in metastatic prostate cancer. There is less prostate cancer invasion when SOD3 is overexpressed, or the expression of the cysteine/glutamate transporter is decreased. This decreased invasion was accompanied by higher extracellular hydrogen peroxide levels and was reversed by overexpression of catalase, which turned hydrogen peroxide into water and oxygen .As discussed above, SOD2 is thought to protect prostate cells under normal conditions. Because radiation therapy can kill cancer cells by generating free radicals, and because SOD2 is a primary antioxidant enzyme, SOD2 can mitigate the effects of radiotherapy. In the presence of radiation, SOD2 is upregulated by the NF-κB pathway and is radioprotective and antiapoptotic ,65. SOD2A study by Josson et al. demonstrated that RelB induced SOD2 in PC-3 cells in response to radiotherapy . This leThis adds to the literature that supports increased concentrations of SOD in cells does appear to confer radioresistance. Several studies utilized the SOD2 gene in vivo and in vitro to confer radioresistance ,73,74,75The radioprotective effect of SOD2 may be limited to oxygen-rich environments. A study from Urano et al. used cDNA to transfect tumor cells with SOD2. Cell lines included a low SOD line, a high SOD line, and two control lines. In the presence of oxygen, both cell lines containing SOD had higher levels of survival after radiotherapy than the control cell lines. In the absence of oxygen, the SOD cell lines were more radiosensitive than the control cell lines. It is important to note that in the absence of oxygen, SOD2 reduced the tumorigenicity of the cell lines containing SOD and did not confer significant radioresistance .Although cells have numerous antioxidant mechanisms, some are more radioprotective than others. A study by Sun et al. created three mammalian cell lines which overexpressed SOD1, SOD2 and glutathione peroxidase, respectively. These cell lines and a control cell line were then irradiated. The cell line which expressed SOD2 provided the most radioprotection followed by the cell line that expressed glutathione peroxidase. Overexpression of SOD1 did not significantly alter the radiosensitivity of the cells .By utilizing our knowledge about the NF-κB pathway and SOD, clinical interventions could be used to fight prostate cancer. A study by Ismy et al. demonstrated that PC-3 cells undergo more apoptosis after administration of exogenous SOD2 . This ocOther studies have focused on decreasing the radioresistance of prostate cancer cells. SN52 is a cell-permeable peptide that blocks the noncanonical NF-κB pathway. It works by inhibiting the binding of p52, and thus the translocation of RelB-p52 into the nucleus. SN52 blocks both baseline constitutive SOD2 expression and radiotherapy-induced SOD2 expression. It was found to increase radiosensitivity of prostate cancer cells when given prior to a 2 Gy fraction  .Even previously known substances have proven to be useful to improving radiotherapy. Pretreatment with 1-alpha, 25-dihydroxyvitaminD (3) suppresses increased levels of RelB caused by radiation therapy in prostate cancer cells expressing the vitamin D3 receptor. This in turn causes reduced expression of SOD2 and increased radiosensitivity of prostate cancer cells .There are some clinical results in humans. ATN-224 is an oral SOD1 inhibitor which has also been shown to have a variety of antitumor activities, including antiangiogenic activity, inhibition of epidermal growth factor receptor and platelet-derived growth factor receptors, and prevention of translocation of RelA into the nucleus. A noncomparative Phase II trial administered high-dose and low-dose ATN-224 to patients with biochemically recurrent hormone naive prostate cancer. At six months, 59% and 45% of patients in the low- and high-dose arms, respectively, were biochemically recurrence free . Of courMutations which result in prostate cancer can originate from ROS-mediated damage. The SOD family is one of the body’s primary antioxidant mechanisms that confers protection from these mutations. Once prostate cancer has developed, SOD level can vary throughout the course of prostate cancer development. Generally, however, the level initially decreases and then gradually increases as the tumor progresses.Unfortunately, in response to therapy, including radiotherapy, SOD, and especially SOD2, can be upregulated. When it is, it confers radioresistance. This is mediated by the NF-κB pathway, which utilizes both the canonical and the noncanonical pathways. However, the noncanonical pathway has been shown to be particularly significant in the induction of SOD2 in prostate cancer. This knowledge provides the unique opportunity to investigate methods that would block SOD2 in prostate cancer but not in normal tissues. Early, promising, translational developments have been made already that utilize our knowledge of the NF-κB pathway and SOD family to improve the treatment of prostate cancer.


Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the progressive degeneration of motor neurons, leading to muscle atrophy, paralysis and even death. Immune disorder, redox imbalance, autophagy disorder, and iron homeostasis disorder have been shown to play critical roles in the pathogenesis of ALS. However, the exact pathogenic genes and the underlying mechanism of ALS remain unclear. The purpose of this study was to screen for pathogenic regulatory genes and prognostic markers in ALS using bioinformatics methods. We used Gene Ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, gene set enrichment analysis (GSEA), and expression regulation network analysis to investigate the function of differentially expressed genes in the nerve tissue, lymphoid tissue, and whole blood of patients with ALS. Our results showed that the up-regulated genes were mainly involved in immune regulation and inflammation, and the down-regulated genes were mainly involved in energy metabolism and redox processes. Eleven up-regulated transcription factors  and one down-regulated transcription factor (NOG) in the nerve tissue of patients with ALS likely play important regulatory roles in the pathogenesis of ALS. Based on construction and evaluation of the ALS biomarker screening model, cluster analysis of the identified characteristic genes, univariate Cox proportional hazards regression analysis, and the random survival forest algorithm, we found that MAEA, TPST1, IFNGR2, and ALAS2 may be prognostic markers regarding the survival of ALS patients. High expression of MAEA, TPST1, and IFNGR2 and low expression of ALAS2 in ALS patients may be closely related to short survival of ALS patients. Taken together, our results indicate that immune disorders, inflammation, energy metabolism, and redox imbalance may be the important pathogenic factors of ALS. CEBPB, CEBPD, STAT5A, STAT6, RUNX1, REL, SMAD3, GABPB2, FOXO1, PAX6, FOXJ1, and NOG may be important regulatory factors linked to the pathogenesis of ALS. MAEA, TPST1, IFNGR2, and ALAS2 are potential important ALS prognostic markers. Our findings provide evidence on the pathogenesis of ALS, potential targets for the development of new drugs for ALS, and important markers for predicting ALS prognosis. Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by progressive muscle paralysis, which involves the degeneration of upper motor neurons (UMN) and lower motor neurons (LMN) in the motor cortex, brainstem, and spinal cord . ALS is After years of research, many new discoveries have been made regarding the pathogenesis of ALS. Various environmental factors have been proposed to be associated with ALS, such as smoking, low antioxidant intake, intense physical exercise, head trauma, inflammation, poor nutritional status, and exposure to electromagnetic fields ,7,8. ImmSOD1 mutations [C9orf72 gene [C21orf2 gene increases the risk of developing ALS [C9orf72 [Abnormal expression of many genes has been shown to play an important role in the occurrence and development of ALS. Superoxide dismutase 1 (SOD1) is a small protein that contributes to partial resistance to oxidative stress . Approxiutations . Additiof72 gene . TAR DNAf72 gene . The half72 gene . Mutatiof72 gene . A genomping ALS , has bee[C9orf72 . As ALS In this study, we used Gene Ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, gene set enrichment analysis (GSEA), and expression regulation network analysis to investigate the function of differentially expressed genes (DEGs) in the nerve tissue, lymphoid tissue, and whole blood of patients with ALS. We identified 12 transcription factors that play important regulatory roles in the pathogenesis of ALS and four prognostic genes that affect the survival of ALS patients. This is highly significant for the development of targeted treatment and prognostic tools for ALS in the future. Thus, our study linked DEGs in ALS, GO, KEGG, and GSEA analysis, and an expression regulation network, which will help to develop new targets for the treatment of ALS. Data were sourced from Gene Expression Omnibus (GEO). The data were RNA expression data obtained from samples harvested from nerve tissue , lymphoid tissue, and whole blood. Detection platforms and sample sizes are shown in Using Pearson’s correlation coefficient, we assessed the correlations of DEGs among the datasets . As the p < 0.05 for nerve tissue, and fold change ≥1.2 and p < 0.05 for lymphoid tissue and whole blood. We screened for DEGs in each dataset. The chip data were processed using the student t-test and modest t-test by the limma R package, and the union of the result obtained by both methods was selected. The sequencing data were processed using edgeR and DEseq2 R package, and the union of the data processed by both methods were selected. The thresholds for DEGs were as follows: fold change ≥1.3 and p < 0.05 in each dataset were considered to be significantly enriched. Second, the DEGs (defined as sel2g) that were regarded as differential expressed in at least two datasets were identified for subsequent analysis. These DEGS were consistently up-regulated (sel2g.up) or consistently down-regulated (sel2g.down). Thereafter, these common DEGs were subjected to GO analysis  and KEGG analysis, using the sel2g data. In addition, functional enrichment analysis, comprising GO BP and KEGG analysis, of the intersection of sel2g and DEGs in lymph or whole blood tissues were performed.First, the DEGs in each dataset were divided into up-regulated genes and down-regulated genes, which were enriched and analyzed respectively, and the enriched KEGG pathways with http://www.pathwaycommons.org/pc/home.do, June 15, 2021). Subnetworks were identified from the networks of up- and down-regulated genes using the Markov cluster algorithm (MCL) clustering function [Expression regulation network analysis was performed to screen for important regulatory factors  among the DEGs in the nerve tissue of patients with ALS, and to analyze the gene expression regulation relationships between genes. Gene regulation relationship data, i.e., “controls-expression-of”, were sourced from the Pathway Commons database (function .p < 0.05) were selected for subsequent analysis. The differential expressed probsets found in both training and validation datasets were kept, and thus 4770 probes were retained.The data used for ALS biomarker screening were sourced from GEO (accession no. GSE112681). The data were from 397 ALS samples, 645 control samples. All these samples were from the blood. The chip platforms were Illumina HumanHT-12 v3.0 and v4.0. As the data were sourced from two chip platforms, only the same probes were retained. The outliers were removed by principal component analysis (PCA) and partial least square discriminant analysis (PLS-DA), and the batch effect were removed using the SVA R package. The genes that met the criteria for differential expression in ALS compared to control samples (After eliminating 35 samples (due to outliers), there were 759 and 248 samples in the training and validation datasets, respectively. The training dataset was used for cross-validation by dividing the samples into two groups: one for training and the other for model evaluation. More precisely, using 5-fold cross-validation, the samples were split into five groups, and four were used for training while the remaining one was used for validation. As the procedure was performed five times, each group had the opportunity to become a training dataset to evaluate the performance of the classifiers. The mean model accuracy was determined across the five groups. The genes associated with high model accuracy were selected as the characteristic genes. The second classifiers were AdaBoost, Random Forest, and support vector machines (SVM). These classifiers were constructed with the identified characteristic genes and their classification accuracy in the training dataset was evaluated. The classification accuracy of the SVM classifier was highest. However, whether there was a difference in both groups (ALS vs. Control) was not considered when initially identifying the characteristic genes. Therefore, the SigFeature R package was used to further screen for characteristic genes with differences in both groups (ALS vs. Control) and strong classification performance in the second classifiers. Finally, the constructed ALS biomarker models were evaluated based on accuracy, precision, recall, and receiver operating characteristic (ROC) curve analysis.p < 0.05). Next, the random survival forest algorithm was used to further screen for markers with the highest correlation with survival (relative importance >0.5). Four key survival-related markers were obtained and then used to construct a multivariate Cox regression model. The regression coefficients of each marker were obtained. The following risk score formula was constructed to determine whether each patient was in the high- or low-risk group: 0.13 × ALAS2 + 0.17 × TPST1 + 0.17 × MAEA + 0.35 × IFNGR2. The risk score for each patient in the training dataset was calculated. The patients were classified according to the median risk score in the dataset. Patients with a risk score lower than the median were included in the low-risk group, and those with a risk score higher than the median were included in the high-risk group. Survival curves were drawn to compare the survival status between the low- and high-risk groups. In addition, a cluster heatmap and boxplots were used to show the expression of the four survival-related genes in the low- and high-risk groups.The data used for survival analysis were from 290 patients in the training dataset and 94 patients in the validation dataset. First, we used univariate Cox proportional hazards regression to screen for survival-related markers . In addition, the up-regulated genes in whole blood were mainly related to phagosome, Th1 and Th2 cell differentiation, NF-κB signaling pathway, and TNF signaling pathway, while those in lymphoid tissue were mainly related to the NOD-like receptor signaling pathway C. FigureAs our results showed that most DEGs were in the nerve tissue of ALS patients, we performed GO and KEGG analysis of the up- and down-regulated genes in the nerve tissue of ALS patients. Regarding the GO BPs, the up-regulated genes were mainly related to proteolysis, macromolecule catabolic process, protein modification and translation, negative regulation of cell differentiation, and negative regulation of cell cycle A. The GOWe performed GSEA to analyze the function of the DEGs in the nerve tissue of ALS patients. KEGG analysis revealed that GSEA positive was mainly related to immunity, inflammation, and cell phagocytosis , and GSEA negative was mainly related to oxidative phosphorylation and neurodegenerative diseases (such as Parkinson’s disease and Huntington’s disease) A. GO BP We analyzed the DEGs co-existing in the nerve tissue, lymphoid tissue, and whole blood and found that there were few of them A,B. FirsTo further screen for transcription factors that may play important regulatory roles in the occurrence and development of ALS, we analyzed the expression regulatory relationships among the DEGs in the nerve tissue and screened for important transcription factors. According to the number of target genes of the transcription factors and the multiple of differential gene expression, we identified 12 important transcription factors . An exprPCA and PLS-DA were performed. PCA revealed that it was difficult to separate the samples between the ALS and control groups A. PLS-DAHeatmap analysis of the training dataset showed that 50 characteristic genes were highly expressed in most ALS patients, while 29 were weakly expressed in most ALS patients . Heatmapp < 0.05)  . Next, w < 0.05) . Next, w < 0.05) A. We als < 0.05) B. Next,  < 0.05) C,D. The  < 0.05) E–H. p < 0.1 for the risk score, indicating that it may be useful for independently predicting the survival of ALS patients regardless of the value of other factors  that are extremely important for the pathogenesis of ALS and four key prognostic markers  . Neuroinflammation is one of the most typical features of ALS. All STAT family members  are related to autoimmune diseases . Signal REL encodes the NF-κB subunit c-Rel. The NF-κB/REL transcription factor family plays a central role in the initiation and regression of the inflammatory response [response . There aresponse . NF-κB iresponse . NF-κB sresponse . Knockouresponse . FUS mayresponse . Taking Runt-related transcription factor 1 (RUNX1) is a highly conserved transcription factor that regulates embryonic development, angiogenesis, hematopoiesis, immune response, and inflammatory response . RUNX1 iSMAD family member 3 (SMAD3) is a key mediator of the typical TGF-β signaling pathway and plays an important role in TGF-β1-mediated transcriptional regulation . PhosphoThe transcription factor CCAAT enhancer-binding protein β (CEBPB) can be activated by a variety of inflammatory stimuli, including IL-17 and LPS, and it regulates many genes involved in inflammation . It is aGA-binding protein (GABP) is a widely expressed ETS family transcription factor that is composed of two subunits: the DNA-binding GABP α subunit and the transactivation GABP β subunit . GABP reNoggin (NOG), which is a member of the TGF-β superfamily, is a bone morphogenetic protein (BMP) antagonist that is related to nerve recovery and nerve regeneration . NOG treOur results revealed that high TPST1, IFNGR2, and MAEA expression and low ALAS2 expression were closely related to short survival in ALS patients. Tyrosylprotein sulfotransferase 1 (TPST1) transfers sulfate moieties to proteins that are then secreted, using 3′phosphoadenosine-5′phosphosulfate (PAPS) as the donor , in the IFNGR2 expression increased in mesenteric white adipose tissue and played a role in cancer cachexia-related systemic inflammation [IFNGR2 was significantly up-regulated [IFNGR2, are up-regulated, suggesting that IFNGR2 may be involved in the pathogenesis [The Th1 cytokine interferon (IFN)-γ is a pleiotropic, pro-inflammatory cytokine that functions through two IFN-γ receptors: IFNGR1 and IFNGR2 . IFN-γ pammation . In a BAegulated . Patientegulated . In Alzhogenesis . Our resMacrophage erythroblast attacher (MAEA) encodes a protein that plays a role in erythroblast enucleation and the normal differentiation of erythroid cells and macrophages. It regulates the development of mature macrophages by mediating attachment to erythroblasts and promotes adult bone marrow erythropoiesis by regulating the maintenance of macrophages ,98. MAEA5-Aminolevulinate synthase 2 (ALAS2) is a rate-controlling enzyme of erythroid heme synthesis . ALAS2 kIn this study, we analyzed the function of the DEGs in nerve tissue, lymphoid tissue, and whole blood of ALS patients and screened for biomarkers related to the survival of ALS patients. We found that the up-regulated genes in ALS were mainly involved in the regulation of immune and inflammation-related processes, and the down-regulated genes in ALS were mainly involved in the regulation of energy metabolism and redox processes. We also found 11 up-regulated transcription factors  and one down-regulated transcription factor (NOG) in the nerve tissue of ALS patients. In addition, MAEA, TPST1, IFNGR2, and ALAS2 were greatly related to the survival of ALS patients. High expression of MAEA, TPST1, and IFNGR2 leads to short survival of ALS patients, while high expression of ALAS2 results in long survival of ALS patients. Taken together, our findings provide evidence for the determination of the pathogenic mechanism of ALS, potential targets for new drug development, and new markers for predicting the prognosis of ALS.


Precipitation and temperature are major controls on multiple ecosystem functions in alpine grasslands. There is scant evidence for the interactive effects of temperature and precipitation changes on the ecosystem multifunctionality (EMF) in alpine meadows. To explore the interactive effects of temperature and precipitation changes on the EMF in alpine meadows, we transplanted meadow blocks reciprocally among three altitudes  on the central Tibetan Plateau. Compared with the home sites (control), the EMF has a trend to increase when meadow blocks were downward transplanted  to the high-precipitation sites but decrease as meadow blocks were downward transplanted to the low-precipitation sites. However, the experimental cooling (upward transplantation) consistently reduced the EMF regardless of the precipitation change. The increase of EMF under the experimental warming was closely related to the variation of both plant and soil functions, whereas the reduction of EMF under the cooling was highly correlated with the decrease of plant function. Our results highlight that climate warming effects on the EMF are greatly associated with precipitation changes in the semi-arid alpine ecosystems. Other coexisting species mainly included Kobresia humilis, Kobresia humilis, Androsace tapete, Polygonum macrophyllum, Thalictrum alpinum etc.  on the central Tibetan Plateau . This reIntact meadow blocks (70 cm × 70 cm wide × 40 cm depth) with attached vegetation were cut off for reciprocal transplantation from the plots at 4,650, 4,950, and 5,200 m in September of 2011 . There wPi is the relative importance value (IV) of the ith species calculated as: IV = (relative height + relative cover + relative biomass)/3  and community height (CH) at each quadrat. The species richness (SR) was defined according to the number of plant species. Aboveground plant biomass (AGB) was estimated using a non-destructive sampling method, and the detailed information was described elsewhere . Soil temperature (ST) and soil moisture (SM) at each collar were measured simultaneously with Re by a Time Domain Reflectometer. Top soil samples (0–10 cm in depth) were collected with a soil auger (diameter: 3.0 cm) within each quadrat. Soil samples for measuring soil organic carbon (SOC) and soil total nitrogen (STN) were air-dried at room temperature and sieved (2–mm mesh). Soil samples for measuring soil microbial biomass carbon (MBC) and soil microbial biomass nitrogen (MBN), ammonium nitrogen , diurnal variation  of ecosystem respiration (Re) at 2 h intervals were measured twice a month in 2014 using the opaque chamber of Li-8100 103 automatic soil COij represents Z-score of the ith ecosystem function in the jth plot and N is the total number of ecosystem functions evaluated  in the ecosystem functions and the EMF between the control and transplanted plots was calculated as: Relative change (Δ) = Transplanted – Control. Pearson correlation and a simple linear model (y = a + bx) was used to determine correlations of the relative changes in the ecosystem functions with that of the EMF. All statistical analyses were performed using SPSS 20.0 , and graphics were drawn using the OriginPro 2019 software .One-way analysis of variance (ANOVA) and the Tukey-HSD test was applied to assess the differences in the VC, CH, SR, PD, AGB, ST, SM, SOC, STN, MBC, MBN, The CH at the home sites generally decreased with increasing altitude from 4,650 to 5,200 m, and there were no significant differences in the SR and PD at home sites among three altitudes . HoweverThe SOC, STN, MBC, and MBN at the home sites were significant higher at 4,950 m compared with that at 4,650 and 5,200 m . The NO3altitude . However 5,200 m . Compare 5,200 m . Warmingon sites . Both thme sites . The ST me sites . Howeverme sites . The grome sites . Coolingme sites . Warmingme sites .The EMF index at the home sites varies greatly along altitudinal gradient, with the highest value at the 4,950 m and the lowest value at 4,650 m . The resltitudes . We founltitudes , but onlltitudes . The annltitudes , 5A,C. Altitudes .In alpine grasslands, numerous studies have explored the relationships among plant or soil microbial diversity and the EMF in alpine grasslands and further demonstrated that aboveground or belowground biodiversity generally positively related to the EMF , the National Natural Science Foundation of China , and a grant from Grassland Science Institute, Tibet Academy of Agricultural and Animal Husbandry Sciences (CYS-TC-2021-001).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Background: Neuromyelitis optica spectrum disorder (NMOSD) is an autoimmune demyelinating disease that recurrently relapses and leads to severe disability. The available choices for disease prevention are few or intolerable. Previous studies suggested that telitacicept may provide a promising therapeutic strategy for autoimmune diseases involving B cells. Therefore, this study aims to assess the effectiveness and safety of telitacicept for recurrent NMOSD.Methods: We will perform a single-arm, single-center, open-label, specialist study with a total enrollment of eight participants. The treatment regimen includes plasma exchange three times and subcutaneous injection of telitacicept for 46 cycles, with a total period of 48 weeks. The primary endpoint is the time to first recurrence after enrollment. Secondary endpoints are Expanded Disability Status Scale (EDSS) score, Opticospinal Impairment Scale (OSIS) score, Hauser Ambulation Index, number of lesions on MRI, and changes in visual evoked potential (VEP), optical coherence tomography (OCT) and immunologic status. All adverse events after medication will be documented and investigated.Discussion: This study will explore the safety and effectiveness of telitacicept following plasma exchange regarding the time to recurrence in neuromyelitis optica spectrum disorder (NMOSD) for the first time.Clinical Trial Registration:Chictr.org.cn, identifier ChiCTR1800019427 Neuromyelitis optica spectrum disorder (NMOSD) is a primary B-cell mediated central nervous system autoimmune disease with an incidence of 0.5–10 per 100,000 people , there mCurrently, infusions of high-dose intravenous methylprednisolone (IVMP) and plasma exchange (PLEX)  are commNMOSD is a primary B-cell and antibody-mediated autoimmune disease . The relThe regulatory role of BLyS and APRIL involves both B-cell maturation antigen (BCMA) receptors and transmembrane activator and calcium modulator and cyclophilin-ligand interactor (TACI) , 20. TelStudies have shown the safety and efficacy of telitacicept in B-cell-mediated diseases such as SLE and RA –25. StudOur study is a single-center, single-arm, open-label clinical study and abide by the SPIRIT reporting guidelines . The purConsidering the small and special nature of the study, the sample size will not be analyzed.The potential participants of this study are patients with AQP4-IgG-seropositive NMOSD in the acute phase, with ≥2 recurrences occurring in the past 12 months prior to screening (including the acute phase recurrence). Written informed consent is required for all potential participants recruited prior to screening in Renji Hospital affiliated to Shanghai Jiao Tong University, School of Medicine. After rigorous screening, the patients enrolled in this study are required to meet all the inclusion criteria without satisfying any of the exclusion criteria .Box 1Inclusion criteria1. Meet the diagnostic criteria for NMOSD in the 2015 international consensus; AQP4-IgG-seropositive.2. Age 18–75 years.3. NMOSD in the acute phase; that is, new onset of neurological symptoms or aggravation of existing symptoms within 30 days before screening, lasting for at least 24 hours without accompanying fever.4. NMOSD recurrence (including acute onset during screening): ≥2 recurrences occurring 12 months before screening.5. Effective contraception during the study. 6. Provide written informed consent.Exclusion criteria1. Progressively deteriorating neurological disorders unrelated to NMOSD.2. Active hepatitis or severe liver dysfunction ; HBsAg-positive patients were excluded. Patients with only HBc-Ab positivity are needed for the HBV-DNA quantitative test and are not excluded if negative.3. Severe renal insufficiency .4. Current pregnancy, breastfeeding or plans to become pregnant in the past 48 weeks.5. Participation in any clinical trial drug within 28 days before enrollment or 5 times the half-life period of the experimental drug (for a shorter time).6. History of splenectomy.7. Allergic reaction: history of allergies to contrast agents or human-derived biological products for parenteral administration.8. Severe psychiatric symptoms and the inability to cooperate.9. Unable to undergo magnetic resonance imaging.10. Medication history of rituximab or mitoxantrone 3 months prior to enrollment.11. Considered unsuitable by the investigators.Participants providing written informed consent will undergo screening tests, and the investigators will review the criteria of patient inclusion and exclusion. If participating individuals meet all the inclusion criteria without satisfying any of the exclusion criteria, they will be documented as registered in our study. After written informed consent is obtained, the study will consist of a screening period with a maximum of 2 weeks and a period of treatment for 48 weeks. The patients will be treated with plasma exchange during the first 2 weeks of the treatment period. Afterward, telitacicept will be given weekly for 46 weeks . After rWhen participants are enrolled, they will be given plasma exchange three times, lasting no more than 14 days. Afterward, they will receive telitacicept treatment weekly for 46 weeks. The time lapse from onset of symptoms to first administration of telitacicept is within 17 to 58 days. The interval is at least 48 hours. A dose of 240 mg will be subcutaneously injected once. The injection site can be the thigh, abdomen or upper arm. At least one clinician needs to observe the patients for 30 min, mainly observing the patient's consciousness and vital signs.Telitacicept is a freeze-dried powder injection. The specification is 80 mg for one piece. It needs to be stored and transported at 2°C~8°C. After rehydration, the drug should not be left at room temperature for more than 30 min. Every 80 mg of drug will be dissolved in 1 mL of injection water into an isotonic solution.The patients will be withdrawn from the study if relapse occurs. Except for special situations, the dosage, method of administration and time interval are not allowed to be changed during the study. However, during or after the 12-week visit, the investigator will evaluate the safety of the therapy in the patient. If the investigator considers that the dose needs to be reduced to ensure safety, then the dose can be adjusted. Supposing that a dose started at 240 mg, it can be adjusted to 160 mg for the first time and adjusted again to 80 mg. The minimum dose is 80 mg.9/L; (2) cell count of neutrophils <1.0 × 109/L; (3) cell count of lymphocytes <0.5 × 109/L; (4) count of platelets <75 × 109/L; (5) aspartate aminotransferase (AST) or alanine aminotransferase (ALT) > 3×upper limit of normal (ULN); or (6) hemoglobin <80 g/L. With the approval of the investigator, drug use can be continued if participants meet the following criteria: (1) cell count of leukocytes ≥2.5 × 109/L; (2) cell count of neutrophils >2.0 × 109/L or the count at baseline; (3) cell count of lymphocytes ≥ 0.75 × 109/L; (4) count of platelets ≥100 × 109/L or the count at baseline; (5) AST or ALT < 2×ULN; and (6) hemoglobin ≥100 g/L or the count at baseline. If the conditions listed above occur again after restarting the drug administration, the patient should permanently discontinue the drug. In addition, if any of the following conditions occur during the study, the patient should also withdraw from the study: (1) AST or ALT > 8×ULN; (2) AST or ALT > 5×ULN and the duration of drug suspension exceeds 2 weeks; (3) AST or ALT > 3×ULN and serum total bilirubin concentration>2×ULN; (4) AST or ALT >3×ULN along with fever, rash, nausea, vomiting, fatigue, right upper abdomen pain, or eosinophilia; (5) cell count of leukocytes <1.0×109/L; (6) cell count of neutrophils <0.5×109/L; (7) cell count of lymphocytes <0.2×109/L; or (8) hemoglobin <65 g/L.The drug should be suspended if any of the following conditions appear: (1) cell count of leukocytes <2.0 × 10At each visit and drug administration, compliance instructions will be provided and confirmed by the investigators.Concomitant drugs or therapies will not be applied during either the screening or treatment period. The oral or intravenous administration of glucocorticoids is not allowed; however, intranasal, intraocular, topical or inhaled glucocorticoids will be allowed. Immunosuppressants are also not allowed, such as tripterygium glycosides, cyclosporine, methotrexate, mitoxantrone, cyclophosphamide, azathioprine, tacrolimus, leflunomide, mycophenolate mofetil and teriflunomide. The following concomitant drugs and therapies are prohibited: biological immunosuppressive agents such as rituximab, hematopoietic stem cell transplantation, lymphatic irradiation and immunoglobulin injection.Mecobalamin and vitamin B1 are allowed to be used during the study. In the case of other diseases during the study, drugs that do not affect the efficacy of the telitacicept (such as antibiotics) can be used. However, the reasons for the use of drugs and method of application should be recorded in detail.The primary outcome is time to first recurrence after enrollment. The definition of recurrence needs to meet the following criteria: (1) new nervous system abnormalities or worsening of existing symptoms; (2) symptoms lasting for at least 24 h; (3) over 30 days after the last relapse; and (4) no associated fever, temperature <37.5°C, or without known infection.Secondary outcomes are listed below:EDSS scoreOSIS scoreHauser Ambulation IndexNumber of lesions in MRIChanges in visual evoked potential (VEP)Changes in optical coherence tomography (OCT)Changes in immunologic statusAll adverse events after medication.The EDSS score, OSIS score, Hauser Ambulation Index are evaluated by two professional physicians. Number of lesions in MRI include number of high signal lesions on T2 weighted imaging (T2WI), low signal lesions on T1 weighted imaging (T1WI) and lesions on gadolinium-enhanced T1WI, which are obtained in MRI of the optic nerve, brain and spinal cord. MRI scans are analyzed independently by radiologists at the Department of Radiology at Renji Hospital affiliated with Shanghai Jiaotong University School of Medicine. The latency and amplitude of evoked potential P100 were recorded by VEP. Optical coherence tomography (OCT) is used to measure the thickness of the retinal nerve fiber layer (RNFL), which is obtained with the “Fast RNFL Thickness” protocol. The titers of anti-AQP4 antibody are tested by the method of cell-based transfection immunofluorescence assay (CBA). Other immune tests are titers of IgA, IgG, IgM, complement 3, complement 4 in serum and lymphocyte subsets in the peripheral blood.Adverse events will be graded using the Common Terminology Criteria for Adverse Events (CTCAE V.4.0). Common adverse events are local reactions at the injection site, including mild to moderate erythema and itchy skin rashes. Other potential adverse events include upper respiratory tract infection, bruises and itching at the injection site, soreness and redness at the injection site, urinary tract infection, herpes zoster, mumps, periodontitis, abnormality of liver function, hypothyroidism, fatigue, headache, sore throat, nasopharyngitis, diarrhea, nausea, vomiting, flu-like symptoms, cough, oral ulcer, nasal congestion, palpebral edema, right upper abdominal pain, joint pain, back pain, and perianal abscess.Patients who are enrolled in this study and receive the test drugs at least once, with at least one post-medication evaluation, will be included in the full analysis set (FAS). The per protocol set (PPS) is described as participating individuals in the FAS who comply with the protocol, including the therapeutic regimen and the availability of the primary outcome with no major violation of the protocol. The primary outcome will be analyzed in the PPS. Participants who are administered the test drugs at least once and have recorded security evaluations will be included in the safety set (SS). Safety will be analyzed in the SS.The quartile of time to recurrence, 95% CI and censoring rate will be calculated.t test will be used for statistical analyses of the difference in means before and after administration.The secondary outcomes will be analyzed in the form of simple descriptive statistics, such as the mean and associated 95% CIs, SDs, median, minimum, and maximum. Paired p-value.All adverse events will be analyzed in the SS. The Medical Dictionary for Regulatory Activities (MedDRA) will be used to code the preferred terms (PT) for adverse events and system organ class (SOC). All adverse events will be listed in detail, and the incidence of different adverse events will be calculated. All statistical analyses will be performed using JMP software (SAS V.9.3), with significance set at a two-tailed All data for every participant will be recorded by investigators in case report forms (CRFs). CRFs will be made according to the plan at screening, at baseline, at 4 weeks, 12 weeks, and 24 weeks after baseline, at the end of treatment and if relapse occurs. The researcher will review and sign the CRF when complete. The investigator needs to re-sign the form if the data are modified.Materials such as the research records, informed consent forms and data of all patients will be reviewed every 6 months to ensure the accuracy and integrity of the data.Patients and/or the public were not involved in the design, or conduct, or reporting, or dissemination plans of this research.All information collected in our study will be stored securely in locked file cabinets at the study site. All information of every participating individual will be identified by a coded number without names or other personal identifiers to maintain participant confidentiality. A password will be applied to protect the database.Our study will be performed according to the principles of the Declaration of Helsinki and has been approved by the ethics committee of Renji Hospital affiliated with Shanghai Jiaotong University School of Medicine. Investigators will explain the study by information sheets. Patients who are willing to participate will have enough time to ask questions. All questions will be explained in detail before obtaining written informed consent. All participants will be provided consent forms. Compensation will be given to those who suffer harm from trial participation.The results of this study will be reported in academic conferences and peer-reviewed journals. The paper will be reviewed and approved by all authors before publication.Recently, an increasing number of studies have focused on treatments involving biological agents in patients with NMOSD (8). Few studies have been conducted on the treatment of telitacicept in patients with NMOSD. Compared with available immunosuppressive drugs such as azathioprine and mycophenolate mofetil, telitacicept shows advantages in targeting a very specific molecule. Therefore, some off-target adverse effects may be avoided, and the treatment may be more accurate.The results of this study may be useful for considering the safety of new biological agents in patients with NMOSD. Studies in healthy volunteers have demonstrated the safety and tolerability of telitacicept in a healthy population , 31. RanThe study will also provide preliminary data on the efficacy of telitacicept in patients with NMOSD. Since NMOSD is a primary B cell- and antibody-mediated autoimmune disease, AQP4-IgG plays an important role in the disease . In our In addition, we will provide a detailed description to demonstrate the efficacy of telitacicept, including the EDSS score, OSIS score, Hauser Ambulation Index, number of lesions in MRI, and changes in OCT, VEP and immunologic status. We expect that the review of this protocol may be useful for other researchers to promote future related clinical trials.Nevertheless, the sampling strategy is a limitation and potential source of bias in this study. We will only be able to preliminarily demonstrate the probable safety and efficacy of telitacicept, as this study is a single-center, single-arm, open-label study. Large-sample, prospective, multicenter, randomized controlled studies need to be performed in the future. In addition, the assessment of safety and tolerability will only be based on a 1-year follow-up visit. A longer follow-up period may be needed.In this study, we expect that telitacicept may be safe and effective in patients with recurrent NMOSD; however, large sample, prospective, multicenter, randomized controlled studies are needed for further validation. Overall, as there are few studies on telitacicept treatment in patients with NMOSD, this study will be useful in presenting preliminary data for future study designs.▸ This study will explore the safety and effectiveness of telitacicept following plasma exchange regarding the time to recurrence in neuromyelitis optica spectrum disorder (NMOSD) for the first time.▸ This study is well-designed for the selection of NMOSD patients with a high recurrence risk.▸ A major limitation of this study is the small sample size, which weakens the power of evidence for the effectiveness of telitacicept. As the current low incidence of NMOSD, a small and special study is firstly conducted.▸ Because eligible NMOSD patients are relatively rare, especially those with a recurrent acute phase within 30 days and a second recurrence within 12 months, this study does not include a control group, which may produce bias.The studies involving human participants were reviewed and approved by the ethics committee of Renji Hospital affiliated with Shanghai Jiaotong University School of Medicine (No 2018-088). The patients/participants provided their written informed consent to participate in this study.JD wrote the initial draft of the paper. Y-TG, YH, and YC revised the paper. Y-TG, YC, and YZ designed the study. Y-TG, YC, YD, YZ, W-WL, and YL reviewed all versions of the protocol and contributed to the start of the study. Y-TG, YH, X-GJ, and JD analyzed and interpreted data. YH, X-GJ, JD, NZ, ZW, and H-JY collected the clinical data. M-CG contributed to the drug storage and electronic CRF. All authors have read and approved the final manuscript and agreed to be responsible for all aspects of the research to ensure that issues related to the accuracy or completeness of the study are properly studied and addressed.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


However, surprisingly, the content of phenolic acid showed the highest values in the area with the lowest pollution. The results of antioxidant tests did not completely correlate with the levels of phenolic metabolites, which may be due to the involvement of other active molecules  in the antioxidant machinery.Gemmotherapy represents the most recent therapeutic technique that uses the properties of extracts from fresh meristematic plant tissues, mainly buds and sprouts, by macerating them in ethanol and glycerol. The harvesting time and the location can significantly affect the chemical composition of the buds. Therefore, this work aimed to point out the possible variability in the phenolic content and the antioxidant potential of extracts prepared from commonly grown trees in the Czech Republic. Extracts from buds collected during autumn and spring in three different localities were analysed using UHPLC-MS (ultra-high-pressure liquid chromatography) for the phenols profile. Five tests assays were used for the evaluation of the extract antioxidant potential. The sampling time positively affected the content of total phenols, flavonoids, and phenolic acids. The increased levels of total phenols and flavonoids in localities with high and medium pollution may be the result of the higher levels of NO and SO Phenolic compounds represent common secondary metabolites in vascular plants. They exhibit great structural diversity and play an important role in the defence responses against various environmental stimuli, such as ultraviolet radiation, heavy metal pollutions, or plant–insect interactions. They are present in all plant organs and are therefore an integral part of the human diet. Polyphenols have drawn increasing attention due to their wide distribution in food, beverages, phytotherapeutics, and potent antioxidant properties .Antioxidants are compounds that can delay, inhibit, or prevent the oxidation of oxidizable materials by scavenging free radicals and diminishing oxidative stress. The imbalance between the excess of reactive oxygen/nitrogen species and endogenous antioxidants leads to oxidative stress, and subsequently to the development of chronic degenerative diseases, such as cancer, atherosclerosis, diabetes mellitus, rheumatism, cardiovascular diseases, or inflammatory injury ,2.In recent years, the demand for traditional and alternative medicine products that use the power of substances naturally contained in plants has increased. One such phytotherapeutic method is gemmotherapy, which uses more biologically active substances in the embryonic parts of plants rather than the grown parts of plants. Here, the extracts are obtained from fresh buds and other meristematic tissues  macerated in a mixture of water, alcohol, and glycerol. The composition of such extracts is more complex because, as well as secondary metabolites, it consists of vitamins, proteins, amino acids, growth factors, hormones, or cytokines, and, therefore, acts on the body at various physiological levels, usually more effectively than common herbal tinctures . Due to Betula spp. L.) leaves and other parts, i.e., buds, bark, or essential oil, are traditionally used for healing urinary disorders, skin diseases, or rheumatism. Triterpenes , tannins, and flavonoids, mainly quercetin and hyperoside glycosides, are the main components and showed antioxidant and antimicrobial activity [Quercus L.) bark is widely used in traditional folk medicine due to its antiphlogistic and antimicrobial properties. Leaf, bark, or acorn extracts also showed antioxidant activity [Acer L., commonly known as maple, have been used in traditional medicine. Phenylpropanoids, flavonoids, tannins, terpenoids, and diarylheptanoids are the most abundant and major bioactive constituents showing antioxidant, antitumor, antimicrobial, anti-inflammatory, and antidiabetic activities [Birch , glycerol, Folin–Ciocalteu’s phenol reagent, sodium carbonate anhydrous, hexahydrate aluminum chloride, hide powder, DPPH , ABTS ), PMS (N-methylphenazonium methyl sulfate), NADH (β-nicotinamide adenine dinucleotide reduced), NBT (nitrotetrazolium blue chloride), SNP (sodium nitroprusside), L-ascorbic acid, 2-deoxy-D-ribose, EDTA, hexahydrate iron(III) chloride, hydrogen peroxide 35% (v/v), sulfanilamide, N-(1-naphthyl)ethylenediamine dihydrochloride, TCA (trichloroacetic acid), and TBA (2-thiobarbituric acid) were purchased from Sigma-Aldrich . All of the solvents, reagents, and standards used were of analytical grade. Ethanol 96% .The chemical reagent and reference standards for UHPLC (ultra-high-pressure liquid chromatography), including formic acid, acetonitrile, protocatechuic acid, Acer pseudoplatanus L., Betula pendula Roth, and Quercus robur L., that originated from three different localities with different levels of environmental impact were used. The plant material was collected in December 2019 and in April 2020, when the buds began to open  represented the locality with the lowest pollution impact. The vegetation around the blind branches of the Elbe in Hradec Králové  represented the locality of common pollution. Artificial plantings with occasional rejuvenation around the Opatovice n. Labem  thermal power station were selected as the polluted location. For more details, see Buds of three different trees,  to open . The polv/v ethanol and 25.5% v/v glycerol. After four weeks of cold maceration (in a dark place at laboratory temperature), extracts were filtered  and diluted with the same solution at a ratio of 1:10 and used for the following analyses. The extraction method followed the protocol for bud preparation detailed in the European Pharmacopoeia . The motw/v) sodium carbonate, and 25 µL of Folin–Ciocalteu reagent solution were incubated for 60 min at 45 °C. After cooling, the absorbance was read at 750 nm [The content of total phenols was determined spectrophotometrically  using the Folin–Ciocalteu method with gallic acid as a standard. Briefly, 30 µL of the sample extract, 470 µL of distilled water, 975 µL 2%  and then incubated at room temperature for 60 min. The absorbance was read at 420 nm. The aluminium chloride method, with quercetin as a standard, was used for the determination of the total flavonoids level . The sam−1, nebulizer 35 psi, sheath gas temperature 380 °C, sheath gas flow 12 L min−1, capillary 2500 V, and nozzle voltage 0 V. Selected MRM transitions were followed for each compound: protocatechuic acid , p-hydroxybenzoic acid , caffeic acid , chlorogenic acid , t-cinnamic acid , ferulic acid , gallic acid , p-coumaric acid , salicylic acid , syringic acid , and vanillic acid  [The contents of phenolic acids were determined by UHPLC on Zorbax RRHD Eclipse plus C18 column  (Agilent) with a 6470 Series Triple Quadrupole mass spectrometer (Agilent) (electrospray ionization in negative ion mode) as a detector. Eluents: (A) 0.05% formic acid in water and (B) 0.05% formic acid in acetonitrile were used in the following gradient program: 0–1 min (5% B), 2.0–4.0 min (20% B), 8.0–9.5 min (70% B), and 10.0–11.0 min (5% B). The MS source conditions were as follows: gas temperature 350 °C, gas flow 9 L minsample is the absorption of the solution with extract, and Acontrol is the absorbance of the solution without extract. Bud extracts were assayed by the discoloration of a solution of DPPH· as previously reported  with som+ assay was performed by bleaching the cationic radical ABTS+ as described by Soto et al. [+ and 2.5 mM potassium persulfate was left to stabilize from 12 to 16 h in the dark at room temperature before use. Afterward, the solution was diluted with methanol  until an initial absorbance of 0.70 ± 0.02 was obtained at 734 nm. The sample extract (10 µL) was mixed with 300 µL of prepared working ABTS+ solution in a 96-well plate and incubated for 6 min in the dark at room temperature. The absorbance was measured at 734 nm in a microplate reader . The scavenging effect was calculated using the formula:sample is the absorption of the solution with extract, and Acontrol is the absorbance of the solution without extract.The ABTSo et al. . A solut3 , H2O2 , and deoxyribose  was incubated in a water bath for 60 min at 37 °C. Then, 1.0 mL of 2.8% TCA (w/v) and 1.0 mL of 1% TBA (w/v) were added and heated in a water bath for 15 min at 100 °C. After cooling the absorbance was read at 532 nm. The percentage of deoxyribose degradation was calculated using the formula:sample is the absorption of the solution with extract, and Acontrol is the absorbance of the solution without extract [For the generation of the hydroxyl radicals, the deoxyribose method was used. The mixture of the sample extract (10 μL), potassium phosphate buffer , ascorbic acid , FeClsample is the absorption of the solution with extract, and Acontrol is the absorbance of the solution without extract.Superoxide radicals were generated by the NADH/PMS system according to the previously described procedure . The samsample is the absorption of the solution with extract, and Acontrol is the absorbance of the solution without extract.The activity was determined spectrophotometrically in a 96-well plate reader. The reaction mixtures consisted of the sample extract (100 µL) and 100 µL SNP (20 mM) were preincubated for 60 min at 25 °C under light exposure. Then, 100 µL of Griess reagent was added, and the absorbance was read at 540 nm. The percentage of scavenging effect was calculated using the formula: p < 0.05. Six individual samples were used for analyses of each parameter. Statistica 10.0  software was used for statistical analyses. The comparison of differences in the experiment was based on the one-way analysis of variance (ANOVA) and Tukey’s test at the significance level The overall phytochemical profile of the analysed material, phenolic compounds included, is influenced by several factors, such as the genetic origin of the plant, or time and technique of sample collection, and the processing of the material. In addition, environmental conditions (influence of biotic and abiotic factors) significantly affect the qualitative and quantitative composition of plant materials and thus the possible therapeutic potential of medicinal products ,6. ThereBased on the results shown in Castanea species. The parallel increase in flavonoids, namely quercetin and rutin, and tannins also increased [The changes in metabolite content, with respect to sampling time, may be related to the transition of the buds from the dormant phase to the actively growing phase, where the need for metabolites is different. In birch, the content of hydrolysable tannins  and flavonoid aglicones decreased by up to 90%, depending on the bud transformation to the adult leaf. The content of phenolic acids (mainly hydroxycinnamic acid derivatives) increased . A similncreased . The worncreased  pointed ncreased .−1 FW, respectively) and flavonoids  were observed in bud extracts in the slightly contaminated locality HK. In the case of oak, the highest values were in the clean locality J. The commercially available extracts of the monitored trees alone showed significantly lower values compared to our measurements. In the case of phenols, there were no significant differences between N and RA producers, 9.39 and 9.69 mg g−1 FW for maple, 16.83 and 19.21 mg g−1 FW for birch, and 11.75 and 13.48 mg g−1 FW for oak, respectively. The content of flavonoids in the maple extract of the company N (6.72 mg g−1 FW) showed a similar level as the extracts from the localities J and HK. On the contrary, the values in the RA extract (4.92 mg g−1 FW) were comparable with the locality O, and at the same time significantly lower from the above. For birch, the values in N and RA were significantly lower compared to our extracts, 6.36 and 5.63 mg g−1 FW, respectively. For oak, the commercial extracts showed similar values as HK, namely 3.10 mg g−1 FW for N and 3.08 mg g−1 FW for RA.The monitored trees responded differently to the environmental impact . For mapPhenolic acids can be divided into two classes: derivatives of hydroxybenzoic acid such as gallic acid, and derivatives of hydroxycinnamic acid (HCA), such as caffeic, chlorogenic, or ferulic acid . Here, wp-hydroxybenzoic and salicylic acids) and HK . For oak  and the content of total phenols and flavonoids. However, phenolic acids were positively influenced strongly at the highest sampling locality. Thus, the authors suggest that a greater influence on the seasonal and altitudinal variation, as opposed to ambient UV-B, would have other developmental or abiotic  factors [Another factor influencing the overall composition and contents of metabolites may be the altitude at which the plants grow. Several meteorological factors, such as temperature, precipitation, or light intensity are closely related to the altitudinal gradient of a specific locality. The content of phenolics in elderberry leaves was significantly affected by the altitude. The increase of HCA was not gradual along the altitudinal gradient (200–1050 m) but, nonetheless, their levels were higher at the hilltop than at the foothill. The authors suggest that higher HCA values are connected with higher light intensity  . In Buxu factors . On the  factors . The locFrom the above, it is clear that the overall qualitative and quantitative composition of phenols can be affected by several factors simultaneously. Studies evaluating several variables simultaneously already differ in determining the main factor influencing the level of phenolic substances. Moreover, constitutive genetic differences may affect the physiology of the studied species more than the different environmental conditions ,24,25.Rubus and Ribes from different companies showed a similar percentage of individual bioactive compounds, but the values differed from laboratory-prepared extracts [The values of individual phenolic metabolites in commercially available preparations (N and RA) vary considerably in comparison with our extracts. From the above, as the possible causes, we can state that they are the sampling locality, its altitude, or the genotype of the plant. Producers declare that the plant material comes from clean sites without environmental pollution or directly from their own planting (genotype selection may take place here). Similar to our results, commercial preparations of the same species of extracts .2−·, and NO· tests with the SAT pattern. The results are expressed as the % of radical scavenging. The results are shown in Studies showing the relationship between antioxidant-rich food intake and the occurrence of some chronic diseases have increased the interest in studying natural antioxidants. Some authors recommend that several tests be used in the testing of antioxidant potential that involve various mechanisms of chemical reaction, such as hydrogen atom transfer (HAT) or single electron transfer (SAT) . Therefo2− scavenging activity was recorded in birch extracts, specifically in the HK locality. Locations J and O did not differ significantly. In oak and maple, the highest levels were observed in the HK extracts . Based on the NO· assay, birch and oak extracts showed the highest activity in locality J; 68% and 79%, respectively. For maple extracts, it was the locality O. The scavenging of the DPPH radical was very effective for all trees, more than 90%. Significant differences between localities were observed only for maple from the HK locality, where the overall activity was the lowest, 95.15%. Strong antioxidant activity, more than 90%, was also observed for the ABTS test. The results of the ·OH scavenging test showed lower values, ranging from 85–90% in comparison with previously mentioned tests. Significant differences between localities were found only in maple extracts. Overall, the highest OSalix, and Ribes extracts was monitored even after considerable dilution of the extract (up to 100-fold). Similar to our results, aqueous and methanol extracts of birch leaves showed high antioxidant activity, more than 80%, measured via the DPPH and ABTS test . HoweverAlcea species in the content of total phenolics, flavonoids, anthocyanins, and overall antioxidant activity [Aloe vera plant was monitored. Extracts of plants from the highland and semi-arid zones of northern India possessed maximum antioxidant potential compared to the tropical zones of southern India [The different antioxidant activities between tree extracts may be due to the variability of composition, content, and chemical character of various active compounds, and the synergy between them and other natural substances . The stuactivity . A signirn India . 3) substituents in the molecules. Caffeic acid, most often esterified with quinic acid as in chlorogenic acid (both have two −OH groups), gallic acid (three −OH groups), and ferulic acid (one −OH and one −OCH3), showed substantial antioxidant properties to scavenge free radicals [2− and NO· tests. However, when we focus on the correlation between the content of the mentioned acids and the antioxidant activity within one tree, it is not possible to draw a specific conclusion about the possible influence of the studied locality. Generally, the radical scavenging activity of phenolic acids depends on the number and position of hydroxyl (−OH) groups and methoxy , and the genetic origin of plants can play an important role. From the above, there is a need for larger and longer studies, that take into account several of the above variables.Based on the data, we can conclude that the time of bud sampling positively affected the content of total phenols, flavonoids, and phenolic acids. The influence of environmental pollution, as another studied variable, was variable among the studied trees. Increased levels of total phenols and flavonoids in localities O and HK may be the result of higher levels of NO and SO


Overweight/obesity in childhood is suggested to increase the risk of hypertension later in life. We aimed to assess whether and how body mass index (BMI) trajectories during 6–18 years of age are associated with hypertension in young adulthood (18–37 years) in the Chinese population.  Based on the China Health and Nutrition Survey (CHNS), a total of 1,872 participants who received ≥2 measurements of BMI during 6–18 years and had assessment of blood pressure (BP) in young adulthood were included. BMI trajectories were explored using latent class growth mixture models, and associations between identified trajectories with hypertension in young adulthood were examined by logistic regression analyses.  Five heterogeneous BMI trajectories were identified: the low slow-increasing (20.03%), low moderate-increasing (56.14%), low rapid-increasing (17.04%), moderate-increasing (3.63%), and elevated-decreasing (3.15%) groups. Compared with the low slow-increasing group, another three increasing groups had gradually elevated risk of hypertension, yielding maximally adjusted odds ratio  (OR (95% CI)) of 2.48 (1.39–4.42), 3.24 (1.66–6.31), and 3.28 (1.19–9.08), respectively, whereas the elevated-decreasing group reversed overweight/obesity to normal weight in childhood, rendering its association with hypertension as not statistically significant (OR (95% CI) = 2.74 (0.98–7.65)).  Our study indicates that there are varied BMI trajectories from childhood to adulthood and that an elevated BMI trajectory during childhood is related with an increased risk of hypertension in young adulthood. In contrast, weight loss of children with high initial BMI may mitigate or reverse the risk. Our findings emphasize the importance of BMI continuous monitoring during early life. Hypertension is a global public health problem, affecting one-third of adults in the whole world . It is aA considerable number of studies have indicated that adult hypertension originates in childhood , 11. OveBody massive index (BMI) is widely accepted to assess children's weight status. It should be noted that BMI changes rapidly in childhood . Recent Given the urgent need to prevent hypertension in young adults, the alarming popularity of childhood overweight and obesity in China , and theThe current study is based on surveys of the CHNS that conducted in 1989, 1991, 1993, 1997, 2000, 2004, 2006, 2009, 2011, and 2015, with participants of each wave being followed up overtime , 26. In A total of 21,378 participants who were followed up between 6 and 37 years of age were extracted from the CHNS. Later, 1,108 and 669 participants without BMI or blood pressure (BP) values were excluded, respectively. Afterward, exclusions were subsequently made for 14,610 participants who received <2 measurements of BMI during 6–18 years of age and 3,119 participants who did not have BP assessment during young adulthood (18–37 years). Finally, a total of 1,872 participants were included in the present analysis.The flowchart of the exclusion and inclusion process of our study population is presented in Height and weight in both childhood and early adulthood were measured without shoes and in light clothes. By using a calibrated beam scale, weight was measured to the nearest 0.1 kg in light clothing. Height was measured to the nearest 0.1 cm without shoes by using a portable stadiometer. BMI was calculated as weight in kilograms divided by the square of height in meters. Each adult's SBP and diastolic BP (DBP) were measured on the right arm with the lower edge of the cuff placed ∼25 mm above the elbow after a 10-min seated rest using mercury sphygmomanometers. When measuring BP in childhood, the width of the cuff inflatable bag is at least 40% of the circumference of the upper arm between the olecranon and the acromion, and the length is 80% to 100% of the circumference of the upper arm. The arm is bare, and the cuff of the sphygmomanometer is at the same level with the heart. SBP is recorded when the first time there is a clear flapping sound (Korotkoff Phase I). DBP is recorded when Korotkoff disappears (Korotkoff V period). BP was measured three times at 30-s intervals, and the mean of the three measurements was recorded.2 ≤ BMI < 28 kg/m2 is considered as overweight, and BMI ≥ 28 kg/m2 is defined as obesity [In childhood, the definition of overweight/obesity adopts the reference standards of the Centers for Disease Control and Prevention . Overwei obesity .The LCGM model was used to identify different trajectory patterns of BMI during 6–18 years of age. Due to the continuity of BMI, the censored conventional model was considered appropriate. We firstly compared the Bayesian Information Criterion (BIC) among the model with two to six trajectories, with all functional forms set to a third-order (cubic) equation. As recommended in the literature, the closer the BIC values to zero, the better the model is fitted . When thP ≤ 0.05.Differences in characteristics of the participants according to the five BMI trajectories were tested using the rank-sum test or 1-way ANOVA for continuous variables and the chi-square test for categorical variables, respectively. We calculated odds ratio (OR) (95% confidence levels (95% CIs)) in three logistic models to evaluate the associations between BMI trajectories and risk of hypertension in young adults. Model 1 was unadjusted. Model 2 adjusted for demographic characteristics of the subjects . To control the possible influence of childhood hypertension status, which was not assessed because BP was only measured at single time point within 1 year in the CHNS, Model 3 additionally controlled for mean values of SBP and DBP  during childhood based on Model 2. All statistical tests were performed using SAS software , and differences were considered statistically significant when two-sided 2 for boys and 17.0 ≤ BMI < 19.2 kg/m2 for girls at 6 years old. Therefore, we named low, moderate, and elevated BMI trajectories referring to normal weight, overweight, and obesity at the age of six, respectively. The numbers of participants were 375 (20.03%), 1,051 (56.14%), 319 (17.04%), 59 (3.15%), and 68 (3.63%) for the low slow-increasing, low moderate-increasing, low rapid-increasing, elevated-decreasing, and moderate-increasing groups, respectively.As shown in The low slow-increasing group approximately follows the lower range of normal BMI , so we c2, and 89 (4.75%) and 69 (3.69%) subjects experienced or developed overweight or obesity, respectively. In early adulthood (18–37 years old), 162 (8.65%) participants suffered hypertension. Significant differences in characteristics  at enrollment of the current study among the five trajectory patterns were observed (P < 0.001).The study variables by BMI trajectories are presented in As shown in In this longitudinal study, with 7,703 BMI measurements from 1,872 Chinese children and adolescents, we identified five distinct BMI trajectories from ages 6 to 18 years. We estimated the relationship between participants on different BMI trajectories and the incidence of hypertension in young adulthood and found that compared with the lowest BMI trajectory, upward trajectories were prone to hypertension. However, participants who remitted overweight/obesity to normal weight showed a neutral association with hypertension in early adulthood.Studies have reported that an excessive increase in BMI in childhood can lead to hypertension in adulthood , 32, 33.Besides these ascending trajectories, we also found a descending trajectory that was with neutral association with hypertension. Consistent with our findings, previous cohort studies have suggested that overweight/obesity in childhood but normal weight in early adulthood could alleviate the risk of adult hypertension in blacks, Hispanics, or white non-Hispanics , 35. AnoMeanwhile, it was found that the low moderate-increasing and elevated-decreasing BMI trajectories almost converged around 18 years of age in the current study. Interestingly, the two groups possessed opposite directed trajectories during childhood and led to varied probability of hypertension in young adulthood. This also suggested that it may be insufficient to assess disease risk only by BMI at one single point.Potential mechanisms linking overweight/obesity to hypertension include hemodynamic changes, sympathetic nervous system activation, sodium retention, renal insufficiency, endocrine changes, insulin resistance and vascular dysfunction due to inflammation, and dietary factors , 36. HigOur data were derived from the CHNS cohort study, which is a longitudinal study with subjects from urban and rural areas in multiple provinces in China. It used standardized protocols and rigid quality control procedures, which may provide a valid appraisal of the relationship between BMI trajectories and the risk of hypertension. Nevertheless, there are some limitations of our study that should be considered. First, our cohort only includes Chinese. The BMI trajectory groups identified may not be generalizable to other populations. Second, the number of participants eventually included in the study is relatively small, and studies with larger sample size are needed to verify our results. Third, hypertension is suggested to be evaluated based on BP values from at least three separate office visits for both adults and children. However, BP was only measured at single time point within 1 year in the CHNS, and the status of hypertension for children and adolescents was not assessed. Therefore, our study was unable to exclude children and adolescents with hypertension at baseline. We tried to adjust the mean BP values during childhood as a compensation, which may not be sufficient to control their impact. Fourth, BMI does not necessarily reflect body fat changes and muscle mass that occur with age . Fifth, Our research supports an association between the excess gain in BMI during childhood and increased chances of hypertension in young adulthood. In contrast, participants who reversed childhood overweight/obesity to normal weight in adolescents may mitigate the risk of hypertension. Such findings are of particular interest because childhood, a phase of very rapid growth, constitutes a critical period for the onset of overweight/obesity. Continuous BMI monitoring is essential in identifying high-risk children, and specific public policies and clinical attention should be formulated for this population. Additional research is needed to clarify childhood growth trajectories and their association with adult hypertension, to more effectively prevent overweight/obesity in children and hypertension in adults.


Acute exacerbations of chronic obstructive pulmonary disease (AECOPD) are serious complications that often require immediate intervention in an emergency department (ED). The aim of this study was to investigate the effect of intravenous magnesium sulphate as an adjuvant in the treatment of AECOPD in the ED.In a double-blind, randomized clinical trial, a total of 60 patients with AECOPD presenting to the ED of Imam Khomeini Hospital in Sari, Iran, were included. The study was conducted between September 2016 and February 2018. Eligible patients were randomly allocated into two groups of intervention and control. Patients in the intervention and control groups received intravenous infusion of magnesium sulfate (2 gr) or normal saline over 30 minutes, respectively. For all patients, Borgdyspnea score, forced expiratory volume in one second (FEV1) result and clinical variables of interest were evaluated before the beginning of the intervention, and also 45 minutes and 6 hours after the commencement of intervention.Regardless of time of evaluation, pulse rate (PR), respiratory rate (RR) and Borg score in intervention group was lower than control group. Also, FEV1 and SPO2 were greater in intervention group compared to control group. However, these differences were not statistically significant (between-subject differences or group effect) (p<0.001). The trends of FEV1, SPO2, PR, RR and Borg score were similar between two groups of study .According to the results of this study, it seems that using intravenous magnesium sulfate has no significant effect on SPO2, FEV1, RR, and PR of patients with AECOPD who presented to ED. Chronic Obstructive Pulmonary Disease (COPD) is one of the leading causes of hospitalization, mortality, and increased consumption of medical expenses –2. In thBronchodilators, corticosteroids, oxygen and antibiotics are the standard treatment for AECOPD. Recently, intravenous magnesium sulfate has been suggested as a potential adjunct therapy for AECOPD. It has been previously shown that magnesium can potentiate the bronchodilatory effect of inhaled beta-2 agonist and has beneficial effects on the function of respiratory muscle –9. In adTherefore, considering the paucity of information and conflicting available evidence, the aim of this study was to evaluate the effect of intravenous magnesium sulphate as an adjuvant in the treatment of AECOPD in the ED.Study design and sample: In a double-blind, randomized clinical trial, a total of 60 patients with AECOPD presenting to the ED of Imam Khomeini Hospital in Sari, Mazandaran Province, Iran, were included. The study was conducted between September 2016 and February 2018.Inclusion and exclusion criteria: The inclusion criteria were known clinically diagnosed moderate COPD  and not having diseases that mimic the clinical features of COPD. Patients who required tracheal intubation or noninvasive ventilation on admission to ED, having a history of cardiac disease, not able to cooperate or undergo spirometry and with reduced level of consciousness were excluded from the study. Others exclusion criteria were use of intravenous magnesium sulfate in the past 24 hours, systolic blood pressure <100mmHg, using medications that contain magnesium, presence of hyperkalemia or hypocalcaemia with ECG abnormalities, respiratory rate <16 and using magnesium hydroxide oraluminum-magnesium hydroxide syrups during the past 24 hours.Data collection: Eligible patients were randomly divided into two groups of intervention and control with equal sample size. Randomization was done using a sealed envelope technique with a computer-generated random numbering system with the help of a nurse who was blinded to the study groups. In addition to conventional standard therapy which includes nasal-cannula oxygen therapy (2–3 L/min), inhaled salbutamol and ipratropium (6 puff every 20 minutes until three times), and also intravenous hydrocortisone (100 mg), patients in the intervention and control groups received intravenous infusion of magnesium sulfate  or normal saline (100 ml) over 30 minutes, respectively. In both groups, the study was conducted after receiving standard treatment for COPD exacerbation, within the first 60 minutes and the patients were clinically stabilized. The doses of medications were the same for all patients. All drugs were prepared in syringes in the same size, color, volume, and shape. The syringes were labeled as A or B by a pharmacist and were administered by a nurse, using a peripheral venous catheter. Also, patients were unaware of the type of medication they were receiving.Throughout the study, patients were monitored for possible side effects. Fifteen minutes after the start of the magnesium sulfate infusion, patellar reflex was assessed in patients, and if the patella reflex was diminished or absent, magnesium sulfate was discontinued immediately.Before the beginning of the intervention (T1), and also 45 minutes (T2) and 6 hours after the commencement of intervention (T3), spirometry was performed for all patients . Also, the respiratory rate (RR), pulse rate (PR), O2 saturation (SPO2), presence of cyanosis, and 0–10 Borg dyspnoea scale (0 indicate the absence of dyspepsia and 10 indicates severe dyspnea) were evaluated.Ethical consideration: The present study was conducted after the approval of the institutional ethics committee. The researchers explained the objectives of the study to the participants, and informed consent was obtained from all participants. Also, this study was registered in the Iranian Registry of Clinical Trials Database (IRCT20150315021480N8).Sample size: A priori sample size was calculated using GPower3.1 with the formula for calculation of samples of repeated measures, based on a presumed effect size of 0.2, a statistical power of 80%, and a type I error of 5%. The overall proper sample size was found to be 54 participants. We therefore recruited 60 patients to account for any dropouts.Statistical analysis: Shapiro-Wilk test was used to evaluate whether data were normally distributed. Descriptive baseline characteristics for two groups were tabulated as Mean (SD), or percentages. The independent t-test was used to compare quantitative variables and the chi-square test was used to compare qualitative variables. Using General Linear Model (GLM), FEV1, RR, PR, SPO2 and Borg dyspnoea score were compared between two groups by repeated measurement ANOVA test. Time of evaluation was considered as within subject factor, intervention state (intravenous magnesium sulfate and control) as between subject factor. Data were performed using the SPSS software . For all statistical tests, P value <0.05 was considered as statistically significant.Data sharing: All relevant data and methodological detail pertaining to this study are available to any interested researchers upon reasonable request to the corresponding author.During the study period, 72 consecutive patients with AECOPD presenting to the ED were screened. Eight patients did not meet the inclusion criteria and four patients declined to participate in the study. All the remaining 60 patients were randomly allocated to two groups and completed the study .In the intervention and control groups, 83.3% (n=25) and 70% (n=21) of the patients were females, respectively (P=0.635). The mean age of the patients in intervention and control groups was 62.9±7.3 and 65.8±3.1 years, respectively (P=0.435). 53.3% and 56.7% of patients in intervention and control groups were current cigarettes smoker, respectively (P=0.92). Most of the patients in intervention  and control groups  had disease duration <5 years, respectively (P=0.75). Dyspnea was the most common complaint of patients . Ninety-three percent (n=28) of patients in the intervention group and 90% (n=27) in the control group were concurrently on medical treatment (P=0.86).The present study showed that although intravenous magnesium sulfate improves FEV1 and SPO2, and decreases PR, RR and Borg dyspnea score, compared to placebo, in patients with AECOPD in the ED; the differences were not statically significant. Based on previous evidence, the efficacy of intravenous magnesium sulfate in patients with exacerbated asthma has been confirmed –17. Magn