Cannabis sativa affect the endocannabinoid system was gathered from the literature. SwissTarget prediction and BindingDB databases were used to anticipate the targets for the phytocannabinoids. The genes related to RA were retrieved from the DisGeNET and GeneCards databases. Protein–protein interactions (high confidence > 0.7) were carried out with the aid of the string web server and displayed using Cytoscape. The Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathway analysis was used to perform enrichment analyses on the endocannabinoid–RA common targets. ShinyGO 0.76 was used to predict the biological processes listed in the Gene Ontology (GO) classification system. The binding affinity between the ligand and the receptors was precisely understood using molecular docking, induced-fit docking, and a molecular dynamics simulation. The network pharmacology analyses predicted that processes like response to oxygen-containing compounds and peptodyl-amino acid modification are related to the potential mechanisms of treatment for RA. These biological actions are coordinated by cancer, neuroactive ligand–receptor interaction, lipids and atherosclerosis, the calcium signaling pathway, and the Rap1 signaling pathway. According to the results of molecular docking, in the context of RA, phytocannabinoids may bind to important target proteins such PIK3CA, AKT1, MAPK9, PRKCD, BRAF, IGF1R, and NOS3. This entire study predicted the phytocannabinoids’ systemic biological characteristics. Future experimental research is needed, however, to confirm the results so far.The endocannabinoid system consists of several phytocannabinoids, cannabinoid receptors, and enzymes that aid in numerous steps necessary to manifest any pharmacological activity. It is well known that the endocannabinoid system inhibits the pathogenesis of the inflammatory and autoimmune disease rheumatoid arthritis (RA). To the best of our knowledge, no research has been done that explains the network-pharmacology-based anti-rheumatic processes by focusing on the endocannabinoid system. Therefore, the purpose of this study is to further our understanding of the signaling pathways, associated proteins, and genes underlying RA based on the abundant natural endocannabinoids. The knowledge on how the phytocannabinoids in  Although inflammation is a typical reaction to tissue damage, it can become out of control and cause further problems. Specific soluble pro-inflammatory mediators, including cytokines or chemokines, prostaglandins, and leukotrienes, are produced as a result of an inflammatory reaction . Many acCannabis sativa (family: Cannabaceae) as well as cannabinoid receptors and enzymes that aid in several steps to exhibit any pharmacological activity [Cannabis sativa (family: Cannabaceae) as a traditional medication for entheogenic and recreational uses. Cannabis sativa, however, may be useful in the treatment of pain, dysmenorrhea, muscle spasms, anorexia, etc., according to earlier research [Cannabis sativa [The endocannabinoid system consists of the numerous phytocannabinoids derived from activity ,5,6. It research ,8. Recenresearch . Furtherresearch . Cannabis sativa . The moss sativa . For a ls sativa ,14.RA is a condition with several diagnoses that is primarily characterized by chronic synovial inflammation. The etiology of this illness involves the infiltration of inflammatory cells, the growth of synoviocytes that resemble fibroblasts, elevated levels of proinflammatory cytokines, and matrix metalloproteinases. These substances encourage angiogenesis and osteoclast activity, which damage joints by altering the development of bone and cartilage . The endCannabis sativa used in this investigation were derived according to reports by Morales et al. [https://pubchem.ncbi.nlm.nih.gov/, accessed on 1 December 2022). The targets of the phytocannabinoids were found using the web servers Swiss Target Prediction and BindingDB . In addition, the canonical forms and 2D structures of the phytocannabinoids were retrieved from the PubChem database . The RA targets were derived from two databases: DisGeNET  and GeneCards . The UniProt database  was utilized to standardize the target names into official gene symbols. The keywords “Rheumatoid arthritis” and the organism “Homo sapiens” were used to seek the targets, which are verified by UniProt ID. To obtain the Venn diagram of the shared targets, Venny2.1.0  was used.The abundant phytocannabinoids of s et al. . In addihttps://string-db.org, accessed on 1 December 2022) to create a PPI network using “Homo sapiens” as the species and “highest confidence > 0.7” as the minimal interaction threshold. Additionally, the detached nodes were kept a secret. Cytoscape software was used to visualize these networks. To identify PPI subnetworks and clusters with a high degree of interconnection within the PPI network, we employed the Molecular Complex Detection (MCODE) method (version 1.5.1). As threshold settings, we set the Cytoscape plug-in MCODE’s maximum depth to 100, node score at 0.2, and K-score to 2 [The chosen targets were added to the STRING 11.0 database  was used to predict Gene Ontology (GO) biological processes. The top 20 biological pathways and processes  were selected based on fold enrichment, and they were then displayed as bubble charts with the adjusted p-values, fold enrichment, gene counts, etc. The GO analysis predicted the various biological processes, cellular components, and molecular functions. A network was constructed using the key pathways obtained from the KEGG analysis for the discovery of the important target proteins that play a significant role in the treatment effects of C. sativa [C. sativa in RA, all the information on the important targets and associated pathways was gathered through online software.The KEGG metabolic pathway analysis was used to perform enrichment analyses on Molecular docking studies of 13 phytocannabinoids were done using Glide XP (extra-precision) implemented in the Schrödinger suite. The energy-minimized structures of the ligands were docked with the crystal structure of PIK3CA , AKT1 ((Gene: Akt1) (AlphFold)), MAPK9 ((Gene: MAPK9) (AlphFold)), PRKCD ((Gene: PRKCD) (AlphFold)), BRAF (PDB ID: 6P3D), IGF1R (PDB ID: 5FXS), and NOS3 (PDB ID: 6NH5) with validated grid parameters [The Desmond module of the Schrödinger software was used to simulate the docked protein–ligand complexes in an explicit water environment. Using a system builder panel, an orthorhombic box was constructed. The simulation of the bound ligand–protein complex was performed using a simple point charge (SPC) model of water. The MD simulation was performed using a constant-temperature, constant-pressure ensemble (NPT) at 310 K and 1.013 bar as an atmospheric pressure intended to last 100 ns. The Simulation Interactions Diagram Report of the Desmond software was used to thoroughly examine the results of the molecular dynamics research ,25.Induced-fit docking was carried out on the best binding energy-containing bioactives with their respective receptors by employing the IFD panel of the Schrödinger suite . Here, tThe binding free energy was calculated from the structural information of the MM-GBSA method. These approaches employ the solvent accessibility method, molecular mechanics, and the generalized Born model to evade the free energy simulations’ convolution . The eneHere, the MM-GBSA calculation ,30 was dhttp://www.swissadme.ch/, accessed on 3 December 2022) were used to determine the physicochemical and pharmacokinetic properties of all the phytocannabinoids, which helps to determine the physicochemical significant descriptors and pharmacokinetically important properties of the phytocannabinoids.The QikProp module of Schrödinger suite   and SwisCannabis sativa, were retrieved from the literature [A total of 13 phytocannabinoids, found in terature . A totalThe phytocannabinoids and common targets were added to Cytoscape 3.9.1 for the construction of a network B,C. ThisA total of 4255 terms linked to the therapeutic effects of phytocannabinoids on RA were found as a result of GO enrichment analysis on common phytocannabinoids–RA targets. 9-tetrahydrocannabivarin , and cannabichromene  (Based on the Glidescores of the phytocannabinoids against PIK3CA (PDB ID: 4L23), the top three phytocannabinoids were discovered to be cannabigerol , Δcal/mol) . The phycal/mol) .In the case of AKT1 (Gene: AKT1), the top three phytocannabinoids were discovered to be cannabitriol , cannabichromene , and cannabielsoin . The phytocannabinoids cannabitriol and cannabichromene formed a common H-bond with Asn279. Cannabitriol formed two additional H-bonds with Lys276 and Asp292. The phytocannabinoid cannabielsoin formed H-bonds with Lys158 and Glu278. All three phytocannabinoids exhibited common polar interactions with Asn279 and Thr291. The superimposition of the three phytocannabinoids predicted that they all easily fit into the Leu156, Val164, Ala177, Met227, Tyr229, Ala230, Met281, and Phe438 residues-based shared hydrophobic binding pocket.8-tetrahydrocannabinol . Cannabinodiol formed H-bond with Ile32 and Ser155, whereas the phytocannabinoids cannabichromene and Δ8-tetrahydrocannabinol formed H-bonds with Glu109 and Met111; and Lys55, respectively. All three phytocannabinoids formed common polar interactions with Ser34, Asn114, and Ser155. Through the superimposition of all three phytocannabinoids, it was revealed that they fitted well into the common hydrophobic binding pocket consisting of Ile32, Val40, Ala53, Ile86, Met108, Leu110, Met111, Val158, and Leu168.In the case of MAPK9 (Gene: MAPK9), the top three phytocannabinoids were found to be cannabinodiol , cannabichromene , and Δ9-tetrahydrocannabinol , and Δ9-tetrahydrocannabivarin . All three phytocannabinoids formed two common H-bonds with Val174 and Gln284, while cannabitriol formed an additional H-bond with Leu279. All three phytocannabinoids formed four common polar interactions with Ser173, Hie231, Asn283, and Gln284. It was discovered via the superimposition of all three phytocannabinoids that they all fit comfortably into the same hydrophobic binding pocket that is made up of the residues Val174, Cys175, Cys264, Met266, Leu279, Cys280, Ile282, and Leu286.In the case of PRKCD (Gene: PRKCD), the top three phytocannabinoids were found to be cannabitriol , ΔIn the case of BRAF (PDB ID: 6P3D), the top three phytocannabinoids were found to be cannabigerol , cannabinodiol , cannabichromene . The phytocannabinoids cannabigerol and cannabinodiol formed H-bonds and π-π interactions with Cys532 and Phe595, respectively. All three phytocannabinoids formed two common polar interactions with Thr529 and Gln530. It was discovered via the superimposition of all three phytocannabinoids that they all fit comfortably into the same hydrophobic binding pocket made up of the residues Ile463, Val471, Ala481, Val2, Leu514, Trp531, Cys532, Phe583, and Phe595.9-tetrahydrocannabinol , Δ9-tetrahydrocannabivarin , and cannabigerol . All three phytocannabinoids formed an H-bond with Met1082. They also formed one common polar interaction with Thr1083. It was discovered by the superimposition of all three phytocannabinoids that they all fit comfortably into the same hydrophobic binding pocket made up of the residues Leu1005, Phe1010, Val1013, Ala1031, Leu1081, Met1082, Ile1156, and Met1160.In the case of IGF1R (PDB ID: 5FXS), the top three phytocannabinoids were found to be ΔIn the case of NOS3 (PDB ID: 6NH5), the top three phytocannabinoids were found to be cannabinol , cannabichromene , and cannabigerol . The phytocannabinoids cannabinol and cannabichromene formed one common π-π interaction with Trp178. Cannabichromene formed one H-bond with Ser354, while cannabigerol formed two H-bonds with Cys184 and Pro334. All three phytocannabinoids formed two common polar interactions with Ser226 and Ser354. It was discovered by the superimposition of all three phytocannabinoids that they all fit comfortably into the same hydrophobic binding pocket that is made up of the residues Trp178, Ala181, Cys184, Leu193, Ala227, Pro334, Val336, Met339, Phe353, and Phe473 .The molecular dynamics simulations of protein–ligand complexes are thought to be the best developed method for assessing the structure-to-function relationships of macromolecules in computer-aided drug design. To clarify the cannabinoid’s potency against the six proteins associated with RA, an impartial molecular dynamics investigation was conducted in the current study. The stability and viability of the best docked ligands against all the target proteins were examined using MD simulation studies, which were run for 100 ns. The root mean square deviation (RMSD), the root mean square fluctuation (RMSF), and protein–ligand interactions were generated with the production of 100 ns MD simulations, and their relevance in the solidity of the protein–ligand complexes was thoroughly examined. Protein–ligand interactions and protein–ligand RMSDs were investigated and visualized during this period.For the PIK3CA–cannabigerol complex, their RMSD for the first 50 nanoseconds was more than 2.5 Å, and from 50 nanoseconds on, the ligand started to stabilize with the protein at above 3.5 Å. It was discovered through protein–ligand contact analysis that roughly 50% of the amino acids were responsible for the hydrophobic interactions with cannabigerol. The remaining 50% were engaged in either the H-bond interactions or the water bridges. Hydrophobic interactions with cannabigerol were largely caused by the residues Met772, Pro778, Trp780, Ile800, Tyr836, Ile848, Val850, Met922, and Ile932, of which Met772 and Ile932 were the most accountable at 0.15 ns and 0.20 ns fractions of the interaction time, respectively. Glu849 and Val851 were responsible for the highest H-bond contacts at 0.10 ns and 0.25 ns fractions, while Met772, Ser919, and Asn920 were responsible for forming highest water bridge contacts at 0.20 ns and 0.35 ns fractions.For the AKT1–cannabitriol complex, their RMSD for the first 60 nanoseconds was stabilized at above 4 Å, while for the next 40 nanoseconds, both the ligand and protein RMSDs were slightly fluctuating at 4.8 Å. Through protein–ligand contact analysis, it was discovered that the residues Phe161, Val164, Ala177, Lys179, and Leu181 were highly responsible for forming hydrophobic contact with cannabitriol, among which Val164 and Lys179 were the most accountable ones at 0.3 ns and 0.6 ns fractions of the interaction time, respectively. Glu234 and Asp439 were responsible for the highest water bridge forming contacts at 0.3 ns and 0.5 ns fractions. Lys179 and Asp439 were the H-bonding residues, whereas Glu234 and Asp439 were the residues forming ionic contacts.The protein–ligand RMSD plots for the MAPK9–cannabinodiol complex showed that RMSD was greater than 4 Å for the first few nanoseconds, increased to 12 Å after a while, and then increased to 16 Å before stabilizing there. Through protein–ligand contact analysis, it was found that the residues Val40, Ala53, Leu57, Ile86, Met108, Leu110, Val158, and Leu168 were responsible for hydrophobic contacts with cannabinodiol, among which Val40, Ala53, Val158, and Leu168 were the most accountable ones at 0.40 ns, 0.30 ns, 0.35 ns, and 0.30 ns fractions of the interaction time, respectively. Ile32 and Lys55 were responsible for the highest water bridging at 0.35 ns, while, on the other hand, Ile32, Lys55, Asn114, Ser155, and Asn156 were responsible for H-bonding contacts.The RMSD for the PRKCD-cannabitriol complex was greater than 4 Å for the first few nanoseconds before increasing to 8 Å and then stabilizing there. According to the protein–ligand contact study, Ser173, Gln284, and Lys285 in H-bond interactions at 0.8, 1.0, and 0.8 ns of the interaction period, respectively, accounted for the majority of the protein–ligand connections with cannabitriol. The remaining amino acid residues were in charge of either creating water bridges or creating hydrophobic interactions.The RMSD for the BRAF–cannabigerol complex was greater than 1.6 Å for the first few nanoseconds before increasing to 2.4 Å, then 2.8 Å, and then stabilizing at that value. Trp531, Phe583, and Phe595 were determined to be primarily in charge of hydrophobic interactions with cannabigerol at 0.22 ns, 0.15 ns, and 0.14 ns fractions of the interaction time, respectively, according to the protein–ligand contact study. The majority of the residues either contributed to the formation of water bridges or H-bonds. Among these, Gln461 and Cys532 played a significant role in the formation of H-bonds. Ile463, Glu533, and Gly534 played a significant role in the formation of water bridges.9–tetrahydrocannabinol complex was more than 3.0 Å for the first few nanoseconds, but with time, it increased to 4.8 Å, where it stabilized. Based on protein–ligand contact analysis, it was found that Met1082 was responsible for the highest H-bond contact with Δ9-tetrahydrocannabinol at the 1.75 ns fraction of the interaction time. The amino acid residues Leu1005, Val1013, Ala1031, Met1142, and Ile1160 were mostly responsible for hydrophobic contacts. Leu1005 and Thr1157 were responsible for forming water bridges  to two (in IFD-generated pose). In case of IFD of NOS3 with cannabinol, the number of π-π stacking interactions increased from one  to two (in IFD-generated pose). Additionally, one H-bond interaction with Ala177 was observed from the IFD-generated pose which was absent in the normal XP-mode Glide docked pose. Furthermore, the prime MM-GBSA binding free energies of the best five docked complexes were calculated and are given in the IFD  of all tData generated by QikProp predicted that all the expected attributes for phytocannabinoids fell inside the acceptable range. The SwissADME-predicted data showed that the phytocannabinoids had significant values for the studied attributes, and based on their physicochemical features, they demonstrated drug-like qualities. All phytocannabinoids obeyed Veber’s rule. The phytocannabinoids cannabigerol and cannabichromene failed to pass/satisfy the blood–brain barrier (BBB) filter , but othThe chronic inflammatory condition known as arthritis is characterized by immune-mediated inflammatory sinusitis, which causes hyperplasia, vasculogenesis, pannus development, the deterioration of cartilage and bone, joint deformity, and functional impairment. Many phytocannabinoids, cannabinoid receptors, and enzymes that are part of the endocannabinoid system all contribute to the pharmacological activity of cannabis in different ways. It is well known that the endocannabinoid system inhibits the pathogenesis of the inflammatory and autoimmune disease RA. Due to the abundance of natural phytocannabinoids, the goal of this study was to build on the signaling pathways, associated proteins, and genes that cause RA using integrated bioinformatics and network pharmacology. The general workflow scheme of the present study is depicted in p-value of <1.0 × 10−16. As would be predicted, the proteins interacted with one another quite a bit. The results of the enrichment analysis showed that the proteins were, collectively, at least loosely physiologically interconnected. For the hub PPI, we chose nodes whose median edge count was greater than double that of all other nodes.We gathered 2723 and 4881 predicted targets for phytocannabinoids from two distinct databases, namely GeneCards and the DisGeNET database, respectively. Thirteen phytocannabinoids and 244 common targets were obtained by the intersection of RA targets and phytocannabinoids (with the aid of venny 2.1.0). The PPI enrichment analysis results were reported with 244 nodes, 699 edges, highest confidence (0.90), and the PPI enrichment The results of the PPI network and GO analysis predicted that the key proteins involved in the numerous phytocannabinoids–RA common targets were primarily those involved in the pathways for cancer, neuroactive ligand–receptor interaction, lipids and atherosclerosis, calcium signaling, and RAP signaling. In cancer, there is an abnormality in the signaling of PI3K/Akt/mTOR due to which many types of research have been done to find the inhibitors of PI3K/Akt/mTOR signaling which may act as anticancer agents. RA is an autoimmune-mediated inflammatory disease where the abnormal proliferation of immune cells, macrophages, monocytes, dendritic cells, and synovial fibroblasts may significantly overlap with the abnormal growth of cancer cells. The results of some recent studies in arthritis using PI3K signaling inhibitors suggest that small molecule inhibitor strategies directed at PI3K signaling may be useful for immune-mediated arthritis . The pre9-tetrahydrocannabivarin had the highest Glidescores and the most drug-like qualities, as well as acceptable ADME/T profiles.In light of the current findings of network pharmacology, PIK3CA, AKT1, MAPK9, PRKCD, BRAF, IGF1R, and NOS3 were chosen as the seven key target proteins closely related to the progression of RA. These proteins were further subjected to molecular docking with phytocannabinoids. Negative binding energy indicates effective binding to the target. Lower binding energy suggests a stable ligand–receptor complex better explained as <−5 kcal/mol called good binding activity and <−7 kcal/mol called strong binding activity. Except for AKT1 and PRKCD, the Glidescores of the top three compounds against the other five proteins were in the range of −8.888 to −6.751 kcal/mol. The top three active ingredients were shown to have a strong binding interaction with their respective proteins. It entails unexpected bias from the materiality, which could hasten the anonymous fallacy in in vitro or in vivo studies. However, the results of the molecular docking experiment suggest the potential therapeutic approaches and could guide trials in animals. Network pharmacology is currently used extensively for various traditional medicines, such as Indian traditional medicine, traditional Chinese medicine, Kampo, traditional Korean medicine, etc. Network pharmacology research is being conducted owing to its robust potential to investigate convoluted relationships among multiple targets. Additionally, network pharmacology accelerates the modernization of the Indian traditional system of medicine by usefully illuminating the synergistic effects of biochemical and systemic biology methodologies. Researchers may identify prospective active ingredients for upcoming in vitro and in vivo studies by using molecular docking to lay out an effective binding mode of the active ingredients with disease-related important target proteins. However, a number of the Ayurvedic formula’s constituents can have antagonistic and synergistic effects on the body while being treated because they target the same protein. It is important to consider these implications and to continue to assess them. The findings of MM-GBSA (binding free energy) and IFD support the molecular dynamics findings that the ligand is stable within the protein. One of the finest molecular dynamics results was obtained with the Akt1-cannabitriol complex, despite its Glidescore of −6.035 kcal/mol. Among the seven proteins, the phytocannabinoids cannabigerol, canabichromene, and ΔThe present study evaluated the integrated bioinformatics and network pharmacology to identify therapeutic targets and molecular processes involving the endocannabinoid system in patients RA. The work used a combined bioinformatics and network pharmacology approach to further examine signaling pathways, associated proteins, and genes that may implicate naturally occurring phytocannabinoids in the pathogenesis of RA. The current research findings suggest that the abundant phytocannabinoids can exert their curative effect through a synergistic effect of multi-components, multi-targets, and multi-pathways in the treatment of RA. However, due to the limitation of network pharmacology and molecular docking, a more complete evaluation of the role of the endocannabinoid system in the immune landscape in patients with RA and a comprehensive analysis of patients with RA and normal controls would be necessary. Hence, further evaluation of in vitro and in vivo studies needs to be carried out to validate the current findings, which is the future scope of the present study. The probable mechanisms of phytocannabinoids in rheumatoid arthritis are depicted in The network pharmacology analysis results predicted that the processes such as response to oxygen-containing compounds and peptodyl-amino acid modification are related to the potential treatment mechanisms for rheumatoid arthritis. These biological actions may be coordinated by cancer, neuroactive ligand–receptor interaction, lipids and atherosclerosis, the calcium signaling pathway, and the Rap1 signaling pathway. According to the results of molecular docking, in the context of rheumatoid arthritis, phytocannabinoids may bind to important target proteins such as PIK3CA, AKT1, MAPK9, PRKCD, BRAF, IGF1R, and NOS3. Due to a lack of experimental validation, this study has shortcomings, although it can inspire in vitro or in vivo studies. However, due to the limitations of network pharmacology and molecular docking, a more complete evaluation of the role of the endocannabinoid system in the immune landscape in patients with RA and a comprehensive analysis of patients with RA and normal controls would be necessary. This entire study predicted the phytocannabinoids’ systemic biological characteristics. Future experimental research is still needed to confirm the results so far.


Moreover, no significant difference was found between those with simple and complex febrile seizures in selenium, vitamin D, homocysteine, vitamin B12, and magnesium levels. Conclusions: Elevated levels of selenium, zinc, vitamin D, and vitamin B12 in the blood of children with fevers help to prevent febrile seizures.Objective: Febrile seizure is a complication that makes physicians and families uneasy when detected in children with a high fevers. This study aimed to compare children with febrile seizures and children without seizures in blood selenium, zinc, homocysteine, vitamin D, vitamin B12, and magnesium levels. Materials and Methods: The study group included sixty-one children between the ages of 1–5 who came to the pediatric emergency department with febrile seizure. The control group had 61 children with fever without seizure, who were compatible with the study group in age, sex, and elapsed time since the onset of fever. Blood samples were taken from the patients during their admission. Selenium, zinc, vitamin D, homocysteine, vitamin B12, and magnesium levels were measured, and the data of the two groups were compared. Additionally, patients in the study group had two subgroups, simple and complex febrile seizures, and their parameters were compared. Results: Selenium, zinc, vitamin D, and vitamin B12 levels were significantly lower in the study group than in the control group ( Febrile seizure (FS) is a seizure accompanied by fever (>38 °C or >100.4 °F) due to any focus infection other than central nervous system (CNS) in children aged six months to five years without a history of febrile seizures or CNS anomaly The febrile seizure is the most common type of seizure in children and occurs in about 2–5% of children . Etiologit does not show focal characteristics,it lasts less than 15 min,it does not recur within 24 h.Febrile seizures can be simple and complex FS. FS is simple if:it shows focal characteristics,it lasts more than 15 min,it recurs within the first 24 h .On the other hand, the FS is complex if: Febrile seizures are generally benign, but according to recent studies: (1) they can cause psychiatric diseases and even sudden death, (2) they can cause brain damage if prolonged, and (3) patients have a 14% chance of developing epilepsy within the first 2 years after febrile status . If the Studies were indecisive regarding whether trace elements and vitamin levels in the blood affect the development of FS. Therefore, this study aimed to investigate how trace elements, such as selenium and zinc in the blood, vitamin D, and homocysteine levels influence the development of FS in children with fever. To estimate the required sample size, we used G*Power  to conduThis research had a study group and a control group. The study group included sixty-one children who applied to the Pediatric Emergency Clinic of Kastamonu Training and Research Hospital between October 2022 and January 2023 with FS between the ages of 1–5. The control group had sixty-one children who had a fever but did not have seizures and were similar to the study group characteristics in age, sex, and elapsed time since the onset of fever formed the control group. Children in the study groups were in two subgroups: simple FS and complex FS. There were comparisons between the study group, control group, and two subgroups of the study group in serum selenium, zinc, homocysteine, and vitamin D levels. The criteria to differentiate simple and complex FS are in Seizures resulting from high fever (>38 °C) due to an infection other than CNS were diagnosed with FS . Those wAll blood analyses were performed from the first blood samples taken after the informed consent form was obtained at the time of admission. Selenium level was measured in serum with Perkin Elmer Atomic Absorption method (PinAAcle 900 Z), zinc level with homogeneous colorimetric enzyme technique (SATURNO 300), homocysteine level with chemiluminescence immunoassay (CLIA) method in plasma with EDTA , Vitamin B12 level with Competitive binding immunoenzymatic assay , 25-OH vitamin D level with CLIA method , and complete blood count with Automatic hematological analyzer .p < 0.05. All data were analyzed using SPSS 26.00 . Statistical Analysis: The significance of differences between the means of the continuous variables was determined by the Mann–Whitney U test for non-normally distributed data . PearsonEthical Approval: The study was approved by the Non-Interventional Ethics Committee of Kastamonu University with the decision number 2022-KAEK-107, date 19 October 2022. p > 0.05)  .The children in the study group had the following health condition issues: 33 (54.1%) of the children had simple FS, 28 (45.9%) had complex FS, 68.8% had a family history of FS or epilepsy, 17 children had lower respiratory tract infection (LRTI), 12 children had upper respiratory tract infection (URTI), 10 children had acute tonsillitis, 10 children had acute otitis media (AOM), 8 children had urinary tract infection (UTI), and 4 children had acute gastroenteritis (AGE) . On the p < 0.001). However, there was no significant difference between the two groups in homocysteine (p = 0.990) and magnesium levels (p = 0.787).p < 0.001) and Immature Granulocyte (IG) (p = 0.01) with respect to two groups. The value of RDW was higher in patients with febril seizure while the value of IG was higher in patients without febril seizure. We conducted a study to determine what changes were present in serum selenium, zinc, homocysteine, and vitamin D levels in children aged 1–5 years who experienced febrile seizures. The levels of selenium, zinc, and vitamin D were significantly lower in children with FS than in children without seizures, but there was no significant difference in homocysteine levels. Furthermore, there was no significant difference in serum selenium, zinc, vitamin D, and homocysteine levels in children with simple and complex FS.Selenium is a trace element, and it is the main component of enzymes such as glutathione peroxidase, deiodinases, and thioredoxin reductase with the following critical roles in muscle function, reproduction, tumor suppression, and antioxidant effects, particularly in brain cells . It is aSelenium deficiency had been effective in the pathogenesis of epilepsy in several studies, but its role in FS has not been determined ,18. In t(1)increasing the effect of glutamic acid decarboxylase, a rate-limiting enzyme in the synthesis of an inhibitory neurotransmitter in CNS, GABA (gamma-aminobutyric acid),(2)facilitating the suppression of an excitatory neurotransmitter in CNS, NMDA (N-methyl D-aspartate) by calcium .Zinc prevents excitatory neuronal discharge and seizure by the following two mechanisms: However, increased cytokines during FS reduce serum zinc levels by stimulating the production of metallothionein. Metallothionein accelerates the passage of zinc to the liver . In manyVitamin D acts as both a neurotransmitter and a brain protector in CNS, and the positive effect of vitamin D on epilepsy patients is well-known ,36. Kalup < 0.001), and they suggest that vitamin D could also be used in the treatment of simple recurrent FS. Similarly, Singh et al. [The role of vitamin D in FC remains unclear, although there are limited studies in the literature. In their study, Bhat et al.  found thh et al.  and Motlh et al.  stated th et al.  reportedHomocysteine is a non-essential amino acid synthesized from the amino acid methionine. Lifestyle, diet, medications, and genetic factors can influence homocysteine levels in humans. High homocysteine levels are a well-known risk factor for neurovascular diseases such as epilepsy, dementia, migraine, and cardiovascular diseases . Homocysteine has a toxic effect on CNS through the following mechanisms: DNA damage, triggering apoptosis, and oxidative stress . HoweverVitamin B12 is a water-soluble vitamin that must be taken in the diet, and it is effective in growth and development in humans. The only dietary source of vitamin B12 is animal foods . VitaminAlthough it is thought that low vitamin B12 may be a factor triggering seizures, its mechanism of action is still unclear ,51. Few RDW is a parameter that measures the differences in volume and size of circulating erythrocytes. In other words, the larger the size of the erythrocytes, the greater their heterogeneity . The usaHigh serum selenium, zinc, vitamin D, and vitamin B12 levels prevent the development of seizures in children between the ages of 1–5 in the pediatric emergency department due to fever.


Cannabis sativa, a versatile plant with numerous varieties, holds promising potential for a wide range of biological activity. As raw materials for research, we chose leaves and inflorescences of hemp varieties such as Białobrzeskie, Henola, and Tygra, which are cultivated mainly for their fibers or seeds. The choice of extraction is a key step in obtaining the selected compositions of active compounds from plant material. Bearing in mind the lipophilic nature of cannabinoids, we performed supercritical carbon dioxide (scCO2) extraction at 50 °C under 2000 (a) and 6000 PSI (b). The cannabinoid contents were determined with the use of the HPLC-DAD method. The antioxidant capabilities were assessed through a series of procedures, including the DPPH, ABTS, CUPRAC, and FRAP methods. The capacity to inhibit enzymes that play a role in the progression of neurodegenerative diseases, such as acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and tyrosinase was also studied. The dominant cannabinoids in the extracts were cannabidiol (CBD) and cannabidiolic acid (CBDA). The highest concentration of eight cannabinoids was detected in the Tygra inflorescences extract (b). The most notable antioxidant properties were provided by the Tygra inflorescences extract (b). Nonetheless, it was the Henola inflorescences extract (b) that demonstrated the most efficient inhibition of AChE and BChE, and tyrosinase was inhibited the most significantly by the Białobrzeskie inflorescences extract (b). Multidimensional comparative analysis enrolled all assays and revealed that the Henola inflorescences extract (b) showed the most substantial neuroprotective potential. Cannabis plant material is essential for the development of many economic areas. For instance, the value of the European CBD market in 2020 was estimated at USD 1.7–1.9 billion ,4,5.Cannabis sp. include the development of dietary supplements and medicines containing cannabinoids (mainly cannabidiol (CBD) and Δ9-tetrahydrocannabinol (THC)). Cannabinoids are synthesized in the cannabis plant through a complex series of biochemical reactions from cannabigerolic acid (CBGA) and cannabigerovarinic acid (CBGVA) which originate from olivetolic and divarinic acids  grade) were supplied by Merck . High-quality pure water was prepared using a Direct-Q 3 UV purification system . Iron (III) chloride hexahydrate, 5,5-dithio-bis-(2-nitrobenzoic acid), 2,2-diphenyl-1-picrylhydrazyl, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), neocuproine, 2,4,6-tri(2-pyridyl)-s-triazine, trolox, tyrosinase, L-DOPA, acetylcholinesterase, butyrylcholinesterase, acetylcholine iodide, butyrylcholine iodide, Trizma® hydrochloride, and Trizma® base were purchased from Sigma-Aldrich . Sodium chloride, sodium dihydrogen phosphate, and sodium hydrogen phosphate, were purchased from Avantor Performance Materials . Ammonium acetate (NH4Ac) and methanol were supplied by Chempur . Cupric chloride dihydrate, acetic acid (99.5%), ethanol (96%), and sodium acetate trihydrate were supplied by POCH . Cannabinoids standards: CBD, CBDA, Δ2 extraction process was carried out under 2000 and 6000 PSI at 50 °C with 250 mL of CO2 as marginal pressure values significantly affecting the neuroprotective activity and exhaustive amount of carbon dioxide assessed during preliminary studies  were used. The flow rate was set at 2.0 mL/min, and the column temperature was set at 35 °C. The injection volume was 10.0 µL, the detection wavelength was 228 nm, and the analysis time was 50 min. The retention time of cannabinoids is presented in The cannabinoid profile analysis of the extract was studied with the use of ultra-high-performance liquid chromatography with the diode array detector (HPLC-DAD) validated method  . The detThe antioxidant activity was performed with the use of four assays: DPPH, ABTS, CUPRAC, and FRAP. Each assay was preceded by the screening of the extracts’ antioxidant activity with descending concentrations of the extracts. Trolox antioxidant activity was studied at a suitable concentration range to inhibit radicals (DPPH and ABTS) or to perform redox reactions (CUPRAC and FRAP). A linear regression equation was established to relate the trolox concentration with its corresponding scavenging percentage (DPPH and ABTS) or absorbance (CUPRAC and FRAP), subsequently, the results presented as mg trolox/g plant material were calculated through the equation, according to the antioxidant properties of the extracts in all four assays ,44. The oA is the absorbance of the control sample and iA is the absorbance of the test sample. The DPPH assay is based on mixing the radical solution with a hydrogen atom-donating substance, as it causes the vibrant violet color to vanish, leading to the formation of the reduced form . The inhThe other measurement that determined the scavenging radicals potential was the ABTS assay in which green cation radicals are produced by the loss of electrons by the nitrogen atoms of ABTS, caused by potassium persulfate. The inhibition of ABTS•+ was calculated using the following formula:A0—the absorbance of the control;A1—the absorbance of the sample.As a first method of determining the reducing potential of an oxidant, CUPRAC was performed. During this assay, phenolic groups of antioxidants are oxidized to quinones, and the neocuproine and copper (II) ion complex (bluish) is reduced to the neocuproine and copper (I) ion complex (yellow). 3+ ion to Fe2+ with the formation of a dark blue complex with 2,4,6-tris(2-pyridyl)-1,3,5-triazine (TPTZ)) by the extracts was the FRAP technique. The last antioxidant technique that determined the reducing properties  to elucidate and interpret the interrelationships between the compound profiles and their influence on the biological activity of the extracts. This analysis was carried out using PQStat v.1.8.4.140 software . The Pearson matrix was also calculated with PQStat software v.1.8.4.140. To identify the extract exhibiting the most robust potential neuroprotective activity, which will include both antioxidant activity  and the capacity to inhibit AChE, BChE, and tyrosinase enzymes, a multidimensional comparative analysis (MCA), which compares multi-feature objects, was performed [The statistical analysis was conducted using Statistica 13.3 software . The data is presented in the form of mean values along with their corresponding standard deviations. The experimental data were analyzed using the skewness and kurtosis tests to determine the normality of each distribution, while Levene’s test was used to assess the equality of variances ,58. To eerformed ,60. SyntCannabis sativa is known to have such properties [Hemp varieties like Białobrzeskie, Henola, and Tygra are primarily cultivated for their seeds and fibers, contributing significantly to various industries. However, a noteworthy aspect often overlooked is the potential use of their leaves and flowers for disease prevention, given their studied and hoped-for biological activity. These plant materials, often treated as waste during the production of seeds and fibers, might possess disease-preventative potential if they possess biological activity. For example, it would be beneficial to present their antioxidant and neuroprotective activity as operties ,62,63,64operties ,66,67. Moperties . By reco2 is a commonly used extractant in the pharmaceutical and natural products industries [2 at 50 °C under 2000 (a) and 6000 PSI (b)  .As expected, the cannabinoid profiles differed between the studied hemp strains  . In addiThe antioxidant properties of the extracts were studied based on two mechanisms: the first mechanism scavenges free radicals: 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), and the second inhibit oxidation reactions by being oxidized themselves: cupric reducing antioxidant capacity (CUPRAC), and ferric reducing antioxidant power (FRAP). 2. The most potent antioxidant activity against the DPPH radical was obtained by the Tygra variety, with the result 0.92 ± 0.03 mg trolox/g plant material and 0.93 ± 0.03 mg trolox/g plant material for the leaves and inflorescences, respectively. Both of the results were statistically significantly better than every other extract. During the DPPH assay , the 1,1Another technique that determined the radical scavenging potential of the extracts was the ABTS method . The ABT2+ ion was shown by Tygra in the florescences (b) extract—8.34 ± 0.01 mg trolox/g plant material, which was statistically greater than every other extract. The ability to reduce cupric ion was also studied (CUPRAC assay) , and theThe last antioxidant model used in this study was FRAP, which determined the ability to reduce ferric ions . The strThe oxidative stress mechanism entails the creation of reactive oxygen species (ROS) through normal cellular metabolism, exposure to environmental toxins, or the presence of conditions that cause oxidative stress. Oxidative stress arises when there is a disturbance in the equilibrium between the generation of ROS and the body’s protective antioxidant mechanisms .Cannabis sativa L. plant material was extracted with methanol and acetone by ultrasound-assisted extraction; subsequently, both of the extracts were tested for antioxidant properties with the use of the ABTS and DPPH assays [Cannabis sativa L. cultivars [Cannabis sativa Futura 75 variety was extracted with ethanol and administered to the mice intragastrically (1.6 mg of cannabis extract/g/day) via stomach tube [3-induced increase in the level of malondialdehyde in the liver and brain and prevented the decrease in catalase activity. Previous studies indicate that antioxidant activity is related to the level of polyphenols in plant material, but the level of cannabinoids contributes significantly to antioxidant activity by Cannabis sativa. In the present study, the most potent antioxidant activity was related to the greatest CBD level. There are also other studies in the literature presenting the antioxidant potential of the cannabis plant. In a study by Mastellone et al., the fiber-type H assays . MethanoH assays . In the ultivars . The totach tube . The extIn our previous research, which inspired the research presented in this paper, we confirmed the antioxidant potential of methanolic, ethanolic, and isopropanol extracts of hemp fiber varieties. The greatest antioxidant power was also associated with the highest concentrations of CBD . HoweverAChE and BChE are enzymes that break down the neurotransmitter acetylcholine in the synaptic cleft. The inhibition of these enzymes can lead to an accumulation of acetylcholine, which can enhance cholinergic neurotransmission and potentially provide neuroprotective effects. Cholinergic neurotransmission plays a crucial role in cognitive function , includiAll of the leaf and inflorescence extracts showed inhibition of AChE . In geneCannabis sativa extracts and the overall results are similar to those of the AChE inhibition, as among the leaves the strongest inhibitory potential was assessed for the Tygra leaves (b)—8.17 ± 0.09 mg galantamine/g plant material , which showed exceptional inhibition properties—166.73 ± 1.87 mg azelaic acid/g plant material and was not significantly different from the Henola inflorescences extract (b) 170.92 ± 5.57 mg azelaic acid/g plant material. The greatest tyrosinase inhibitor and most statistically different from all other extracts was the Białobrzeskie inflorescences extract (b)—212.22 ± 8.83 mg azelaic acid/g plant material.In a Kim et al. study, hemp seeds ethanol crude extract was fractioned with hexane, dichloromethane, ethyl acetate, and butanol . TyrosinIt would be beneficial to extend the investigations to include experiments conducted on cell lines. After receiving promising results, it would be suggested to expand the research to in vivo animal studies to gain insights into the potential neurodegeneration-preventative applications of the extracts.PCA was used to explain the variation in the data in the present study, namely, to verify if and how the sum of the eight cannabinoids’ content impacts the biological activity of the extract. Factor 1  accounted for approximately 73.9% of the observed variation in the samples, whereas factor 2  explained approximately 16.13% of the variation . The ABTThree antioxidant assays: ABTS, FRAP, and CUPRAC, showed a strong correlation with some of the cannabinoids and the strongest with CBDA—0.941, 0.774, and 0.885, respectively . AChE anThe Tygra inflorescences extract (b) exhibited the highest antioxidant potential among the tested extracts. The most effective inhibition was observed in the case of AChE and BChE by the Henola inflorescences extract (b). The inhibition of tyrosinase was most pronounced in the case of the Białobrzeskie inflorescences extract (b). As the result of multidimensional comparative analysis, the Henola inflorescences (b) extract was determined as one with substantial neuroprotective potential, which comprises antioxidant activity and the ability to inhibit enzymes related to neurodegeneration.Patients’ preferences have undergone significant changes, reflecting a growing demand for natural and alternative remedies ,104. HemAll the tested varieties of hemp  showed the potential for antioxidant and neuroprotective properties, expressed by the possibility of scavenging radicals, reducing ferric and cupric ions, and inhibiting esterases and tyrosinase, but the greatest neuroprotective effect was found in extracts from Henola hemp inflorescences obtained as a result of extraction with carbon dioxide at 50 °C under and under 6000 PSI. Due to the promising results, the Henola variety holds the highest potential for effective use in neurodegeneration prevention, while also contributing to the reduction in associated agricultural waste.Bearing in mind all the advantages of supercritical carbon dioxide extraction in the face of the confirmed neuroprotective effect of the leaves and inflorescences of the tested hemp varieties, this method of obtaining preparations/semi-finished products should be recommended as the most optimal and safe in the development of products containing hemp varieties such as Białobrzeskie, Henola, and Tygra.


Many low- and middle-income countries are now shifting toward diets that are higher in added sugars, unhealthy fats, salt, and refined carbohydrates. Childhood obesity and chronic diseases have all been linked to unhealthy food consumption. Despite this, the majority of Ethiopian infants and children consume unhealthy food. There is also a scarcity of evidence. Therefore, the objective of this study was to assess the prevalence of unhealthy food consumption and its associated factors among children ages 6–23 months in Gondar City, northwest Ethiopia.A community-based cross-sectional study was conducted from June 30 to July 21, 2022, in Gondar city. Multistage sampling was used to select 811 mother-child pairs. Food consumption was measured through a 24-hour recall. Data were entered into EpI Data 3.1 before being exported to STATA 14 for further analysis. A multivariable logistic regression analysis was employed to identify the factors associated with unhealthy food consumption. An adjusted odds ratio (AOR) with a 95% confidence interval was used to show the strength of the association, while a P-value of 0.05 was used to declare the significance of the association.The percentage of children with unhealthy food consumption was 63.7% . Maternal education , living in an urban residence , GMP service , age of the child 18–23 months , and family size of more than four  were significantly associated with unhealthy food consumption.In Gondar City, nearly two thirds of infants and children received unhealthy food. Maternal education, urban residence, GMP service, child age, and family size were all significant predictors of unhealthy food consumption. Thus, improving the uptake of GMP services and family planning services is critical to reducing unhealthy food consumption. Many low- and middle-income countries (LMICs) are going through a nutritional transition, with diets shifting toward more added sugars, unhealthy fats, salt, and refined carbohydrates . As a reGlobally, 40 (5.9%) million children under the age of five are overweight or obese, with the majority of these children living in low- and middle-income countries . CentralChildhood obesity has consequences for adulthood obesity, which can lead to psychosocial problems as well as cardiovascular risk factors such as high blood pressure, high cholesterol, diabetes, and sleep disordered breathing –18. In aReports show that food marketing environments , maternaEthiopia has various initiatives and programs in place to minimize all forms of malnutrition, including overweight and obesity, as part of its national development plan since children under the age of two have been identified as being part of the first key window of opportunity . HoweverA community-based cross-sectional study was conducted from June 30 to July 21, 2022, in Gondar City, Northwest Ethiopia. Gondar City is the capital city of Gondar city administration and the central Gondar zone, which has six sub-cities. It is 727 km from Addis Ababa, the Federal Democratic Republic of Ethiopia’s capital, and 120 km from Bahir Dar, the capital of the Amhara National Regional State. The city has 3 hospitals, 8 health centers, and 14 health posts. The city has a total population of 454,445 people, including 22,945 children under the age of two.All infants and young children aged between 6 and 23 months, with their mothers or caregivers, living in Gondar city were considered the source population, whereas all infants and young children aged between 6 and 23 months, with their mothers or caregivers, in selected Kebeles living in Gondar city were considered the study population.Based on the following assumptions, a sample size was calculated by using the single population formula: Because there have been no previous studies, a 95% CI was calculated using 50% of the previous prevalence, a 5% margin of error, and a 10% non-response rate. Finally, a total of 845 samples were obtained with 2% design effects. To identify study participants, a simple random sampling procedure was used. A list of infants and children was found in health posts. First, two sub-cities were selected from the total number of sub-cities in Gondar by using the lottery method. Second, six Kebeles were selected from each of the selected sub-cities by using the lottery method. Third, after having the number of all 6–23-month-old children from the selected kebeles, corresponding to the month of the 6-23-month-old children, samples were allocated proportionally to each kebele. Finally, a total of 845 participants were taken from each kebele through a simple random sampling technique.The data collection instruments were structured questionnaires that were pretested. The questionnaire was adapted from the World Health Organization (WHO) Infant and Young Child feeding (IYCF) indicators and other literature with some modifications –32. The Two-day training was given on the contents of the questionnaire, interviewing technique, the purpose of the study, and how the principal investigator approaches and maintains the respondents’ confidentiality. A pre-test was conducted with 5% of the total sample size. The quality of the data was checked every day after data collection, and any errors were corrected before the following data gathering measures. Prior to analysis, the data was cleaned up and cross-checked. The principal investigator and the supervisor closely monitored the process throughout the data collection period and made any necessary corrections.To determine each child’s unhealthy food consumption (UFC), the mother was asked to list all foods consumed by the child in the 24 h preceding the survey, such as Juices, soda, coffee or tea with sugar, candies, chocolate, cakes, sweet biscuits, ice cream, potato chips and instant noodle. Children who got at least one food from the above lists were classified as meeting the unhealthy food consumption; otherwise, they were considered to be getting healthy food consumption .A mother with less than two-year-old children who read a newspaper or magazine, listened to the radio, or watched television at least once per week is considered to have satisfactory media exposure; children who did so less than once per week are considered to have unsatisfactory media exposure.Postnatal care (PNC) is defined as care provided to the mother and her newborn baby immediately following placental birth and for the first 42 days of life .GMP utilization was measured by taking a child’s GMP and having them participate in GMP at least once in the previous three months.The household wealth index was determined using principal component analysis (PCA) by considering household assets, such as quantity of cereal products, house, livestock, and agricultural land ownership. First, variables were coded between 0 and 1. Then, the variables were entered and analyzed using PCA, and those variables having a communality value of greater than 0.5 were used to produce factor scores. Finally, the factor scores were summed and ranked into poor, medium, and rich wealth index .Cleaned and coded data was entered into EPI Data Version 3.1 and exported to STATA Version 14 for further analysis. Descriptive and summary statistics were presented in the form of text, tables, and graphs. Both frequency and tables are used to summarize descriptive statistics. Variables with a p-value less than 0.2 in the bivariable analysis were fitted into the multivariable logistic regression analysis. A multivariable logistic regression analysis was employed to identify the factors associated with unhealthy food consumption. An adjusted odds ratio (AOR) with a 95% confidence interval was used to show the strength of the association, while a P-value of 0.05 was used to declare the significance of the association.A total of 811 mother-child pairs with children aged 6–23 months were enrolled in the study, with a response rate of 96%. The mean age of the respondents was 32.33 years, with a standard deviation of 6.24. More than one-third of the mothers, 277 (34.1%), were between the ages of 25 and 30. More than three-quarters of the respondents, 631 (77.8%) and 608 (75%), were married and Orthodox Christians, respectively. More than one third, 283 (34.9%), of the mothers were housewives. Less than a quarter, 182 (22.4%), and 160 (19.7%) of the mothers and fathers, were able to read and write their educational status, respectively. Housewives made up less than half of the 349 mothers (43%) , of the children were within the age range of 12 to 17 months. The majority of study participants, 685 (84.5%), lived in urban Kebeles. More than half, 485 (59.8%) of respondents, had fewer than or equal to four family members. Most 724 (89.3%) of the participants did not have a garden at home. More than two-thirds 620 (76.40%) of the study participants, received advice on child feeding practices from health professionals. More than two thirds, 624 (76.94%) of the participants, began providing their children with complementary feeding, and 604 (74.5%) of the respondents had satisfactory media exposure. The vast majority of children, 730 (90%), are currently breastfed (Table ).Nearly two thirds, 494 (60.9%) of the respondents’ mothers, had four and above ANC visits during their last pregnancies. Almost three-quarters 579(71.4%) of the mothers received information on infant feeding during any of their antenatal care (ANC) visits. The majority, 720 (88.8%) of the study participants, had PNC visits during their last pregnancies, and nearly two-thirds of 500 (61.7%) of the children had growth monitoring and promotion (GMP) in the last three months. Regarding the birth place, the majority 773, (95.3%) of children were from health facilities .The multivariable logistic regression revealed that educational status of the mother, age of the child; urban residence, family size, and GMP service were all significantly associated with unhealthy food consumption Table .Mothers unable to read and write were 1.89 times  more likely to consume unhealthy food as compared to those with a diploma and above.Children aged 18–23 months were 47% less likely  to consume unhealthy food than those aged 6–11 months.Children living in families larger than four were 1.22 times more likely to consume unhealthy food than those living in families smaller than or equal to four .Moreover, mothers who lived in urban areas were 4.55 times more likely  to provide unhealthy food as compared to mothers who lived in rural areas.Mothers who had not received GMP services were 2.07  times more likely to provide unhealthy foods as compared with mothers who had received GMP services.Unhealthy food consumption among children aged 6–23 months in Gondar city was 63.7%. This finding is consistent with a study done in Iran (66.8%) . WhereasMothers unable to read and write were 1.89 times more likely to consume unhealthy food as compared to those with a diploma and above. The possible explanation might be that women who are unable to read or write are unconcerned about the short- and long-term consequences of unhealthy food consumption . In addiChildren living in families larger than four were 1.22 times more likely to consume unhealthy food than those living in families smaller than or equal to four. This could be due to the fact that mothers with larger families take longer to prepare foods and, as a result, purchase unhealthy items for their family. Furthermore, intra-household food distribution may boost the desire to consume unhealthy foods ; unhealtMoreover, mothers who lived in urban areas were 4.55 times more likely to provide unhealthy food as compared to mothers who lived in rural areas. The finding is supported by the global food policy report . The posChildren aged 18–23 months were 47% less likely to consume unhealthy food than those aged 6–11 months. Our finding is contradicted by studies done in Africa, Asia , and NepMothers who hadn’t had GMP visits were 2.07 times more likely to provide unhealthy foods as compared to mothers who had GMP visits. The possible reason might be due to the fact that GMP focuses on empowering mothers to know about and become competent in appropriate child care and feeding practices through individual and group child feeding practices . GMP is The strength of this study was that it was the first of its kind in the study area targeting unhealthy food consumption among children aged 6–23 months. As a limitation, the data collection was done based on past feeding practices, which might be socially desirable. Since the data collection was done in a single 24-hour recall, the finding did not indicate the usual dietary habit of an individual child.In Gondar City, nearly two thirds of infants and children received unhealthy food. Maternal education, urban residence, GMP service, child age, and family size were all significant predictors of unhealthy food consumption. Thus, improving the uptake of GMP services and family planning services is critical to reducing unhealthy food consumption.


Hypercalcemia is a complex medical condition characterized by elevated levels of serum calcium (>10.5 mg/dL) in the bloodstream, often arising from various underlying etiologies. This condition presents a significant clinical challenge due to its diverse clinical manifestations and potential for serious complications. Profiling and understanding hypercalcemia is essential for accurate diagnosis, appropriate management, and improved patient outcomes.In this study, we delve into the comprehensive profiling of hypercalcemia, encompassing its epidemiology, pathophysiology, clinical presentation, and diagnostic approaches. We explore the multifaceted etiological factors contributing to hypercalcemia, including primary hyperparathyroidism, malignancies, granulomatous disorders, medications, and more. We highlight the intricate interplay between parathyroid hormone, vitamin D, and other regulatory mechanisms that influence calcium homeostasis, shedding light on the underlying molecular pathways.Furthermore, we discuss the diverse clinical manifestations of hypercalcemia, ranging from asymptomatic cases to severe, life-threatening complications involving the renal, gastrointestinal, cardiovascular, and neuromuscular systems. Accurate diagnosis is pivotal, and we evaluate the array of laboratory tests, imaging modalities, and specialized assays that aid in identifying the root cause of hypercalcemia. We emphasize the importance of a systematic approach to differential diagnosis and the significance of risk stratification to guide clinical decision-making.The evolving landscape of treatment options for hypercalcemia is also explored, encompassing both acute management and long-term strategies tailored to the underlying etiology. We assess the role of hydration, pharmacological agents, and surgical interventions, underscoring the need for individualized therapeutic plans based on the severity and underlying cause of hypercalcemia.In conclusion, the profiling of hypercalcemia is a multidimensional endeavor that necessitates a comprehensive understanding of its underlying mechanisms, diverse clinical presentations, and diagnostic intricacies. This study intends to serve as a valuable resource for healthcare professionals, offering insights into the complex terrain of hypercalcemia. Hypercalcemia is defined as the serum calcium concentration two standard deviations above the mean values. The normal serum calcium ranges from 8.5 mg/dL to 10.5 mg/dL [Hypercalcemia can be driven by more than 25 different diseases, a number of drugs, and even dehydration. The majority of individuals with hypercalcemia suffer from primary hyperparathyroidism and various types of cancer . Among pSymptoms of hypercalcemia are evident once the serum levels exceed 12 mg/dL which clinically manifests as ‘bones, stones, abdominal moans and physical groans’ . Renal mAfter taking a detailed history, and obtaining serum calcium levels, PTH and Vitamin D levels should be estimated to classify the type of hypercalcemia: PTH Dependent or PTH Independent . This caThis prospective study was conducted on adult patients (age > 18 years) with hypercalcemia  diagnosed on two separate occasions at a tertiary healthcare center, from 1st January 2019 to 31st December 2019. A total of 200 patients were enrolled in the study after satisfying the inclusion and exclusion criteria and were followed up for a period of three months. A thorough clinical history  and general examination were carried out on each subject. Thereafter, an etiological evaluation was carried out and they were followed up for a period of three months. A provisional diagnosis guided specific investigations, such as serum protein electrophoresis, tissue biopsies and aspiration cytology, parathyroid assays, thyroid profiles, serum ACE and 25 hydroxy Vitamin-D levels. The etiology, clinical presentation, course of treatment, and outcome of each patient with hypercalcemia were assessed. Wherever necessary, appropriate radio-nuclear imagings were performed. Ethical clearance was obtained from the "Research and Ethical Committee, Dayanand Medical College and Hospital, Ludhiana, Punjab, India" (BFUHS/2K19p-TH/10723).The collected data was analyzed using Statistical Package for the Social Sciences . For quantitative data, the mean and SD were determined, whereas, for qualitative variables, proportions and percentages were computed. Using the Student t-test and the ANOVA test, quantitative factors between the study groups were compared. The chi-square test was used to compare categorical data. Statistical significance was defined as a probability value  less than 0.05.A study enrolled 200 patients with confirmed hypercalcemia between January 2019 and December 2019. The patients were aged 18-83, with 18.3% aged 18-40 and 40.2% aged 41-60. The majority were male (51.5%) and had mild hypercalcemia (41%). Gastrointestinal symptoms were the most prevalent, with no significant difference in renal symptoms. Neurological symptoms increased from mild to moderate cases, with generalized symptoms more common in moderate and severe groups. Musculoskeletal symptoms increased with severity, with the most prevalent in severe cases (41.1%). Generalized symptoms were more common in the moderate (66.1%) and severe groups (60.7%) compared to the mild group (46.3%), showing statistical significance (p=0.045). Musculoskeletal symptoms increased with severity, being the most prevalent in severe cases (41.1%), and the difference was statistically significant (p=0.012), as depicted in Table The hospitalization outcomes of patients diagnosed with hypercalcemia, classified based on the various causes of the condition and the different outcomes: discharge, death on arrival (DOR), discharge against medical advice (DAMA), and death are summarized in Table The relationship between the cause of hypercalcemia and patient outcome at three months follow-up was assessed in 200 patients, out of which 22 died, and 178 survived. Hyperparathyroidism was found in 9.1% of the dead and 24.2% of the survivors p = 0.173), and drug-induced hypercalcemia was found in 9.1% of the dead and 16.9% of the survivors (p = 0.630). Chronic liver disease (CLD) was found in 4.5% of dead patients and 6.2% of survivors (p = 0.533), and thyrotoxicosis was found in 4.5% of dead and 3.4% of survivors (p = 0.603). At the three-month follow-up, only malignancy showed a significant association (p=0.01) with patient outcome, as depicted in Table , and druThe current study was carried out at Dayanand Medical College and Hospital, Ludhiana over a period of 1 year, i.e., 1st January 2019 to 31st December 2019. Consecutive patients from the emergency department with confirmed hypercalcemia on two occasions were enrolled in our study. Patients were followed up at one month and three months intervals respectively. Out of 200 patients, 87 (43.5%) were in the age group of 41-60 years while 85 (42.5%) were in the age group of 61-80 years. The mean age of the cohort was 58.3 years ± 13.6 years (range 18 to 95 years), which was similar to the study conducted by Kuchay et al. (55.2 years ± 17.9) . There wThe majority of our study patients had mild hypercalcemia  followed by moderate  and severe hypercalcemia . Our results were comparable with another Asian study by Lee et al.  who alsoIn our study, there was no significant correlation between gender and the severity of hypercalcemia whereas, in the study conducted by Lee et al. , severe Two patients (9.1%) died of multiorgan dysfunction secondary to pancreatitis in the hyperparathyroidism group in the hospital. The overall prognosis for hyperparathyroidism was found to be favorable, particularly post-parathyroidectomy. One patient (4.5%) died of chronic liver disease (CLD) in the hospital due to decompensation whereas two (9.1%) died of vitamin D toxication in the hospital. One patient (4.5%) had in-hospital mortality due to multiorgan dysfunction secondary to sepsis. One patient (4.5%) with thyrotoxicosis and CLD, died of aspiration of variceal bleed whereas the exact etiology of mortality could not be ascertained in one (4.5%) of patients post-discharge.Malignancy and hyperparathyroidism are the two most common etiologies associated with hypercalcemia in hospital settings whereas in outpatients, hyperparathyroidism is seen more commonly. The majority of the patients with hyperparathyroidism are diagnosed in an asymptomatic stage in Western countries whereas in India, most of the patients are diagnosed when symptomatic . This diThis study had some limitations: Firstly, it was a single-center study and extrapolation of these results on other populations needs to be confirmed. The study had a relatively small sample size and therefore confounding effect of malignancy on outcome could not be studied.In conclusion, the profiling of hypercalcemia is an intricate and multifaceted undertaking that holds paramount importance in clinical practice. Through a comprehensive exploration of its epidemiology, pathophysiology, clinical manifestations, and diagnostic methodologies, we have gained valuable insights into the complexity of this medical condition. Hypercalcemia's diverse etiologies, spanning from primary hyperparathyroidism to malignancies and beyond, highlight the need for a systematic and tailored approach to diagnosis and management. As healthcare professionals, it is crucial to recognize the subtle variations in the presentation of hypercalcemia and employ a meticulous diagnostic strategy to identify its underlying cause. The integration of laboratory tests, imaging studies, and specialized assays enables us to differentiate between benign and malignant forms of hypercalcemia and guides us toward informed clinical decisions.Malignancy and hyperparathyroidism were the most common causes of hypercalcemia in our hospital setting with vitamin D intoxication as an emerging cause of hypercalcemia. Hypercalcemia has varied manifestations therefore hypercalcemia should be included in routine workup. Furthermore, our examination of the profiling of hypercalcemia underscores the importance of a multidisciplinary approach, where acute interventions and long-term management plans are customized to address both the severity of hypercalcemia and its root cause. In our pursuit to optimize patient outcomes, the profiling of hypercalcemia empowers us to navigate through the complexities of this condition. By staying attuned to the latest research, embracing advancements in diagnostic techniques, and tailoring interventions to each patient's unique circumstances, we can effectively manage hypercalcemia and mitigate its potential complications.In summary, the comprehensive profiling of hypercalcemia serves as an indispensable compass, guiding clinicians in their quest to unravel its intricate nature and offering patients the best possible avenues for diagnosis, treatment, and ultimately, improved quality of life.


In children with enthesitis-related arthritis (ERA), the hip and sacroiliac joint function might be impaired if not properly treated. We sought to evaluate the effectiveness of anti-tumor necrosis factor-α (TNF-α) therapy using the inflammatory indicators, Juvenile Arthritis Disease Activity Score 27 (JADAS27) and magnetic resonance imaging (MRI).We conducted a single-center retrospective study of 134 patients with ERA. We evaluated the effect of anti-TNF therapy on the inflammatory indicators, active joint count, MRI quantitative score, and JADAS27 over 18 months. We used the Spondyloarthritis Research Consortium of Canada (SPARCC) and the Hip Inflammation MRI Scoring System (HIMRISS) scoring systems for hip and sacroiliac joints scoring.n = 87, 64.93%). There were no differences in HLA-B27 positivity between the biologics and non-biologics treatment groups . Children who received anti-TNF  therapy improved significantly. Children with ERA used DMARDs and biologics at baseline (Group A) were followed up to 18 months, and their active joint count , JADAS27  and MRI quantitative scores (P = 0.001) were significantly lower than those at baseline. Some of the patients  were treated with DMARDs at the onset of the disease, but did not show significant improvement (Group B). After 6−18 months of switching to anti-TNF therapy, related indicators of the children were significantly lower than at baseline and 1 month (P < 0.013). At 18 months, a total of 33 patients  in Group A and 7  in Group B reached inactive state.The average age of onset of children with ERA was 11.62 ± 1.95 years, and they were treated with disease-modifying antirheumatic drugs (DMARDs) combined with biologics (Eighteen months after diagnosis, anti-TNF therapy was found to be effective in children diagnosed with ERA. MRI is important for the early diagnosis of juvenile idiopathic arthritis. TNF-α inhibitors can significantly improve the clinical manifestations of sacroiliac joint and hip involvement in patients with ERA. Overall, the real-world study provides more evidence for precision diagnosis and treatment for other hospitals, families and patients. Enthesitis-related arthritis (ERA) is a form of juvenile idiopathic arthritis (JIA), defined by the International League of Associations for Rheumatology (ILAR)  to prima2.This study is a single-center clinical retrospective analysis and involving 134 children diagnosed with ERA from January 2018 to March 2021 in our institute, and who met the newly revised JIA classification and the ILAR diagnostic criteria from 2004. ERA is closely associated with IBD and/or subclinical gut inflammation. According to the norms, calprotectin tests was prescribed for all the ERA patients and gastroscopy and colonoscopy examination were subsequently conducted for those with high faecal calprotectin. IBD and/or subclinical gut inflammation were excluded from our study. The long-term treatment and follow-up of all patients were conducted by rheumatologists. All children with ERA were followed up five times for ≥18 months and all children with ERA underwent MRI examination at baseline, 1, 6, 12 and 18 months as the inclusion criteria. Demographics, clinical characteristics, laboratory data, treatment effect, and MRI quantitative scores data from onset to 1, 6, 12, and 18 months were collected and analyzed. Patients of 6–17 years who fulfilled the ILAR criteria rather than the new PRINTO standard were deemed eligible. As demonstrated by clinical data, ERA often involves both the sacroiliac and hip joints at the time of diagnosis. The inclusion criteria for sacroiliac joint involvement were: (i) occult pain in the lower back, buttocks, proximal thighs, and groin and (ii) morning stiffness, albeit usually lasting <30 min. The clinical manifestations of hip involvement were (i) spontaneous groin, thigh, and hip pain with or without a history of trauma; (ii) limited internal and external rotation of the symptomatic hip(s) after the initial clinical assessment; and (iii) both acute and chronic inflammatory changes on MRI .Demographic profiles of newly diagnosed children with ERA, individual and family history of inflammatory and autoimmune diseases or psoriasis, time from symptom onset to diagnosis, site of joints involved, active and restrictive joint count, acute uveitis, joint effusion, medication, and related laboratory tests, such as C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), HLA-B27, platelet count, alanine aminotransferase, and aspartate aminotransferase, were recorded and analyzed. Serum CRP level and ESR were measured at baseline and at 1, 6, 12, and 18 months. Joint lesions can show bone marrow edema (BME), bone erosion, and joint effusion on MRI. During follow-up, each patient's medications, such as NSAIDs , disease-modifying antirheumatic drugs (DMARDs) , and TNF-α blockers  were recorded. At the same time, rheumatologists can comprehensively evaluate the disease activity in children by assessing the CRP, ESR, MRI quantitative score, active joint count, physician and parent visual analog scale score, and JADAS27. Acute-phase reactants are objective measures of inflammatory response. Among them, ESR is more sensitive for the detection of disease activity, while other inflammatory response indicators including CRP, white blood cell, and platelet counts are not sensitive .Quantitative assessment of the treatment changes in bone marrow edema of children's sacroiliac and hip joints by MRI, often use the Spondyloarthritis Research Consortium of Canada (SPARCC) and the Hip Inflammation MRI Scoring System (HIMRISS).During the study, children with ERA who received biological agents from baseline to the end of the follow-up were included in Group A, while those who were initially treated with non-biological agents but did not show significant improvement were included in Group B.This study was approved by the Ethics Committee of Children's Hospital of Nanjing Medical University (202008041-1).2.1.This scoring method is based entirely on the assessment of an increased signal on T2 with fat suppression or short tau inversion recovery (STIR) sequences denoting bone marrow edema on oblique coronal slices of the SI joint. All signal changes within the iliac bone and sacrum up to the sacral foramina were scored on six consecutive slices through the SI joint. These slices were selected based on the SPARCC protocol, which defines the acquisition characteristics and encompasses most of the synovial compartment. The sacral interforaminal bone marrow STIR signal forms a reference for the determination of increased signals in the SI joints. Each SI joint was divided into four quadrants, and the presence of an increased STIR signal in each of these four quadrants was recorded in each of the six slices, giving a maximum score of 48. The presence of a lesion exhibiting either an intense signal  or depth of ≥1 cm anywhere within each SI joint of the six slices was given an additional score , bringing the total score to 72 .2.2.The HIMRISS was used to assess the changes on MRI from baseline to 18 months. MRI was performed with a coronal STIR sequence with a slice thickness of 4 mm and a field of view of 400  ×  400 mm. The baseline and follow-up MRI examinations of both hips had to be performed every 6 months during the 18-month follow-up period. The MRI scans at baseline and at 18 months were calculated using the HIMRISS. The HIMRISS protocol was used to score the inflammatory changes on the MRI of the hip joints, including the BME of the femoral head and acetabulum, and synovitis , 16.2.3.P < 0.013.This retrospective, self-controlled study adopted a single-sample design. The data in this study were verified to conform to a normal distribution. A Paired T-test was used to assess the differences in remission of clinical indicators  and MRI improvement between different follow-up time points , and use Bonferroni test correction, which makes the data more accurate and meaningful. Measurement data are expressed as mean ± standard deviation (x ± s), and enumeration data as frequency and percentage. Statistical analyses were performed using IBM SPSS 23.0 . Statistical significance was set to 3.3.1.n = 74), 43 patients (58.11%) with ERA were HLA-B27 positive. In Group B (n = 13), 6 (46.15%) children with ERA were HLA-B27 positive, with no significant difference (P > 0.05). There were only five cases of acute uveitis and 15 cases of inflammatory back pain. In addition, of the children with joint pain, 104 had one or more joint effusions.We recruited 134 children with ERA from January 2018 to March 2021, and treatment follow-up was carried out for 18 months. The baseline demographic characteristics of the children diagnosed with ERA are shown in 3.2.P = 0.000), and ESR  in Group A, were significantly lower between each visit and the baseline (P < 0.013). However, in Group B, at one month of follow-up, ESR  and CRP  did not improve significantly or worsened compared with baseline. Subsequently, the treatment of ERA children in Group B was converted to anti-TNF, and at 18 months of follow-up, CRP  and ESR  were significantly lower than that at baseline. There were no significant differences in leukocytes, platelets, alanine aminotransferase, and aspartate aminotransferase during follow-up (P>0.013). Laboratory measures declined with the treatment. This indicates that biological agents can improve the inflammatory indicators of children with ERA.During the 18-month follow-up period, all enrolled patients underwent clinical symptom assessment and MRI examination. Laboratory tests, such as CRP  and hip  decreased significantly compared with baseline. In Group B, the children were transferred to biological agents due to the poor treatment effect of non-biological agents in January. As treatment progresses, the MRI quantitative scores of sacroiliac joints (1.92 ± 2.37) and hip joint (0.62 ± 1.21) were significantly lower than at baseline  and 1 month   . Therefo3.4.In principle, the selection of treatment follows the ILAR guidelines. The medication plan can be timely adjusted according to the examination results to improve the treatment efficacy and promote individualized treatment.P < 0.013). Thirteen children (9.70%) were administered oral NSAIDs combined with DMARDs at the initial stage of the disease according to disease activity and inflammatory response. However, within one month of follow-up, there were 13 children whose inflammatory markers, MRI quantitative scores, active joint count, JADAS27, CRP, ESR and TNF-α increased or did not improve significantly. Therefore, on the basis of the treatment with cDMARDs, etanercept  and golimumab  were added. After the 18 months follow-up, the disease activity and MRI quantitative scores of the affected children were significantly improved compared with the baseline (P < 0.013). Taking JADAS27 as the comprehensive evaluation criterion, at 12 months of follow-up, a total of 8  patients in Group A and 3  in Group B reached the state of disease inactivity. At 18 months, a total of 33 patients  in Group A and 7  in Group B reached inactive state. The symptoms and inflammatory index of 9 patients improved significantly after treatment with biological agents. However, MRI showed that BME was relieved one year after treatment with a TNF inhibitor. Symptoms, inflammatory indices, and MRI examinations of three cases treated with cDMARDs and bDMARDs showed no significant improvement from baseline to 18 months.In children with ERA who presented to our hospital, after completing the relevant inflammatory indicators  and MRI examination, the results showed high inflammatory indicators and poor MRI results; thus, conventional disease-modifying antirheumatic drugs (cDMARDs) combined with biologic disease-modifying antirheumatic drugs (bDMARDs) were usually chosen as the preferred treatment plan. A total of 87 children (64.93%) were treated with biologics, 13 of 87 children with ERA were treated with anti-TNF when they were followed up to 1 month, and the remaining (35.07%) with non-biologics. Of those treated with anti-TNF inhibitors, 71 received etanercept, 13 adalimumab, 2 golimumab, and 1 infliximab. During treatment, 11 children (8.21%) switched to other TNF-α inhibitors. After 1-, 6-, 12-, and 18-month treatment, the clinical symptoms, laboratory indicators, active joint count, JADAS27, and MRI quantitative scores results of Group A improved, without serious adverse reactions  and southern (18.8%) Europe and the lowest in Latin America (6.4%), Africa and the Middle East (5.9%), and Southeast Asia (5.0%) (.1% and sP < 0.013).Laboratory tests are essential for assessing affected children's situation and disease activity. Previous research confirmed that CRP, ESR and TNF-α are the main indicators of disease activity in JIA as well as the key evaluation indicators in JADAS27. Our data showed that CRP and ESR decreased from baseline at all stages, from the initial diagnosis to follow-up, with significant differences . Sometimes, children with ERA do not have symptoms such as joint pain and limitation of motion, but inflammatory reactions in the joints can still be found during MRI re-examination. Therefore, MRI is a key auxiliary examination method for early detection, diagnosis, and treatment.MRI changes in the inflammation of the sacroiliac and hip joints, including BME, were assessed in patients with ERA using SPARCC and HIMRISS. BME is a sensitive, but relatively nonspecific parameter. Pathological studies have shown that the cause of BME is the infiltration of inflammatory cells into the bone marrow, which reflects the pathological changes in osteitis and is regarded as an early marker of inflammation. This is supported by the evaluation of the activity of related clinical diseases and the concentration of markers (ESR and CRP) in the acute phase . CompareThe current first-line drugs for the treatment of children with ERA are NSAIDs. DMARDs therapy has been widely accepted as a second-line drug for JIA for children with limited mobility, mild bone marrow edema, and fluid accumulation on joint MRI. Methotrexate has good efficacy and safety in the treatment of JIA, and is currently the most widely used DMARD for the treatment of JIA . StudiesRecombinant human TNF receptor type II antibody fusion protein is an anti-TNF biological agent; the most commonly used biological agents are etanercept and adalimumab. Several studies have recently confirmed that TNF inhibitors are effective in children with ERA , with reIn recent years, the application of recombinant human type II TNF receptor antibody fusion proteins in children has been widely accepted , 26. In Recently, the study of fecal calprotectin (fCAL) levels in children with ERA has gradually entered the relevant research field. The study found that among patients with various subtypes of JIA, the fCAL concentrations were highest in those with the ERA subtype , alongsiIn summary, ERA accounts for a large proportion of patients with JIA in the Asian population. Therefore, it is necessary to formulate effective diagnostic criteria and personalized treatment schemes for children with ERA. Besides, due to the small sample size, there is no statistical analysis on the efficacy of different treatment schemes, so it is necessary to carry out multi center retrospective research. This real-world study is a description and summary of the clinical data of children with ERA in our hospital, so this study not only confirms the effectiveness of tumor necrosis factor inhibitors in children with ERA, but also provides more evidence for precision diagnosis and treatment for other hospitals, families and patients.