This study first reports the two-staged, double benefits of PD + MAO, and demonstrates its potential in clinical applications for inhibiting peri-implantitis, especially in patients with severe infection risk.Peri-implantitis are a major problem causing implant failure these days. Accordingly, anti-infection during the early stage and subsequent promotion of osseointegration are two main key factors to solve this issue. Micro-arc oxidation (MAO) treatment is a way to form an oxidation film on the surface of metallic materials. The method shows good osteogenic properties but weak antibacterial effect. Therefore, we developed combined strategies to combat severe peri-implantitis, which included the use of a novel compound, PD, comprising dendrimers poly(amidoamine) (PAMAM) loading dimethylaminododecyl methacrylate (DMADDM) as well as MAO treatment. Here, we explored the chemical properties of the novel compound PD, and proved that this compound was successfully synthesized, with the loading efficiency and encapsulation efficiency of 23.91% and 31.42%, respectively. We further report the two-stage double benefits capability of PD + MAO: (1) in the first stage, PD + MAO could decrease the adherence and development of biofilms by releasing DMADDM in the highly infected first stage after implant surgery both   •in vitro.PAMAM-DMADDM shows toxicity reduction and osteoconductive properties •Micro-arc oxidation treatment shows osteoconductive but weak antibacterial effect.•PAMAM-DMADDM + Micro-arc oxidation treatment shows multifunctional properties. Study sfections ,8 and usfections . Traditifections , whereasfections . Highly promised ,13. Accopromised .Microbial adhesion usually initiated in the early stage of treatment, which is defined as the first four weeks after implant surgery . AggressThere are two main strategies to solve the problem of peri-implantitis: implant material surface modification and local drug treatment ,17. Devein vitro and in vivo have been evaluated in the study.The present study reports a novel biomaterial PD and its combination with MAO to combat peri-implantitis at two stages. PD and MAO both have antibacterial and osseointegration-promoting effects. In this study, a highly infected rat model, which provides a foundation for clinical application, was used to test the multifunction of PD and MAO. The study indicated that local treatment with PD provides a strong antibacterial activity at the high-risk first stage of initial four weeks especially for individuals with severe infections or immunocompromised individuals. In the second stage, MAO implants will mainly have an antibacterial effect and promote osseointegration. Antimicrobial characteristics, anti-infection abilities, and osteogenesis characteristics of PD and MAO treatment 22.12SO4 solution. Subsequently, a 70 V DC voltage for 1 min was applied as previously reported [2O3 particles, and etched in acid solution ; the HA was first sandblasted and then plasma sprayed with about 30 μm of hydroxyapatite; the machined technique was mechanically polished in sequence with grit SiC paper . In this experiment, HA, SLA, and machined titanium act as controls. All discs and rods were sterilized in an ethylene oxide sterilizer .Titanium discs were prepared in the size of 6 mm in diameter and 1 mm in thickness, and titanium rods were prepared in the size of 1.5 mm in diameter and 20 mm in length. The novel MAO implant was made by an anodizing device. The samples were immersed into a 1 M Hreported ,35. Sand2.21H NMR spectroscopy, particle size testing, Zeta potential testing, loading efficiency evaluation, and release experiment in vitro, as well as biocompatibility testing in vivo and in vitro were performed.The same weight (1 g) of PAMAM and DMADDM were mixed in ethanol and stirred at room temperature for 1 h then 55 °C for 3 h. The mixed solution was added to a 1000D dialysis bag, and rotate dialysis for 2 days. Following this, the mixed solution was freeze dried. Then collecting the solid powder and storing at −20 °C for later use. Subsequently, 1H NMR spectroscopy, particle size, and Zeta potential were showed in the Supplementary materials. To calculate the loading efficiency of PD, the known concentration of DMADDM was diluted in a gradient, and ultraviolet absorbance measurement  at a wavelength of 220 nm was used to determine the standard curve. Then the dialyzed fluid was collected, diluted 160 times, and the concentration of non-carried DMADDM was detected via ultraviolet absorbance measurement  at a wavelength of 220 nm. The loading efficiency and encapsulation efficiency of PD was calculated using the following formula:The detailed methods of testing the D is the quantity of DMADDM that was loaded within PAMAM. MP is the quality of PAMAM. MTD is the quality of DMADDM added in the beginning. Ultraviolet absorbance measurement was used to determine the concentration of DMADDM within PD.MFor the releasing experiment, 780 mg PD was added to dialysis tubing and the release experiment was conducted in 100 mL ethanol. Then, 1 mL of the solution was collected for later detection and replaced with fresh ethanol at 1, 2, 3, 4, 5, 6, 12, 24, 36, 48, 72, 96, 168 (1 week), 336 (2 weeks), and 504 h (3 weeks), respectively. The concentrations of the DMADDM were tested by the method mentioned above.2.3S. aureus and C. albicans were diluted to 1 × 106 CFU/mL in the medium [Antibacterial experiment: Titanium was added to a 24-well plate, then added 1 mL RPMI1640 medium  to each well. PD solution at a mass fraction of 0.25 mg/mL and 0.5 mg/mL, or PBS as a control, was subsequently added to the wells. e medium . After 25 cells/mL. All groups were incubated in an incubator at a constant temperature of 37 °C under 5% CO2 for 24 h. Then the medium was refreshed with several kinds of medium: osteoinductive medium (OM), PM, OM + 0.25 mg/mL PD, OM + 0.5 mg/mL PD and OM + 0.5 mg/mL PAMAM. After incubating for 24 h, all the groups except for the PM group were refreshed with OM, while the medium in PM group was refreshed with PM. The medium was changed the next day and the cells were incubated for 4, 7, 14, and 21 days [The effect of PD on preosteoblasts: MC3T3-E1 mouse preosteoblastic cells  were inoculated in the α-MEM medium containing 10% fetal bovine serum . The cells were seeded at a density of 1 × 10 21 days . MTT, AL2.4All the animal experiments were followed by the ARRIVE guidelines 2.0, the details of checklist could be seen in the Supplementary materials S. aureus-C. albicans + 0.5 mg/mL PD, MAO + S. aureus-C. albicans + 0.25 mg/mL PD, and MAO + S. aureus-C. albicans + PBS, which were abbreviated as 0.5 mg/mL PD, 0.25 mg/mL PD, and PBS groups. Briefly, three kinds of solution were prepared at first, the C. albicans + S. aureus + PBS, C. albicans + S. aureus + 0.25 mg/mL PD, and C. albicans + S. aureus + 0.5 mg/mL PD, and both the C. albicans and S. aureus were at a concentration of 106: 106 CFU/mL. Then, MAO Ti rods were immersed in the solutions for 40min, dried in a sterile environment. Subsequently, after the rats were anesthetized, the Ti rods were implanted into the left femurs. For assessment of the cortical bone destruction of femurs, X-ray imaging was used after 1 day and 3 weeks of the implantation (n = 5). And the radiographic scores were evaluated according to the X-ray imaging (n = 5), the details of scoring were showed in previous study and the group names could not be seen when scoring [SPF-grade laboratory animal room, and the water, food and bedding were changed every other day.The anti-infection and osteointegration effect of MAO + PD were assessed by a highly infected rat model. Fourty-five Sprague-Dawley  rats were divided into three groups, MAO +  scoring ,41. To e2.5SPSS, version 22.0  was used to perform the statistical analysis. One-way analyses of variance (ANOVA) were performed to detect the significant effects of the variables at a p-value of 0.05.3staphylococci (approximately two-third), and most of them are by the highly virulent pathogen S. aureus, which accounts for approximately 35.5% infections [C. albicans is a major fungal species in implant-related infections [S. aureus and C. albicans are also often detected together in peri-implantitis [C. albicans and S. aureus are a relatively mature model for studying peri-implantitis; in this study, we evaluated the effect of MAO using this severe infection model [A very large percentage of implant-related infections are caused by fections . C. albifections . S. aurelantitis . The twolantitis . Biofilmon model ,46,47.C. albicans was obviously reduced on MAO, but the amount of S. aureus did not change compared with the other three kinds of titanium discs , MTT assay, and colony-forming unit (CFU) assay. SEM showed that the amount of um discs . Moreoveum discs . Further p＜0.05, , but thet change .S. aureus and C. albicans, the results have shown that the antibacterial effect of MAO is not strong enough to combat severe peri-implantitis. However, MAO titanium showed some antibacterial ability for inhibiting C. albicans : 3.46 , 5.52, 6.03 , 4.6 , and 4.13 , which was similar to the results reported before [δ (ppm): 3.25–3.30 , 2.60–2.62 , 2.80–2.82 , 2.37–2.45 , 3.22–3.27 , and 2.72–2.77 , which was similar to the results showed before [The structures of DMADDM, PAMAM, and PD were tested by d before . And ford before . The peaZeta potential is an important parameter to describe the electrostatic interaction between particles in a dispersed system and the dispersed system under the influence of this electrical phenomenon; furthermore, it is very significant to study the physical stability of the drug dispersed system . The res1H NMR results, the compound PD was successfully synthesized.Particle size is also an important index for evaluating the biological performance of carrier systems . The parDendrimers are a kind of macromolecules which are three-dimensional spheres with branches emanating from the center . The cor3.1.2In vitro hemolysis test is one of the most basic methods to detect blood compatibility and is also one of the important indices to detect the safety of biomaterials [Hemolysis is the release of hemoglobin through the rupture of red blood cells or through partially damaged membranes. aterials ,64. The aterials . Therefoin vitro, and is often used as a cell model for bone metabolism research [MC3T3-E1 is a pre-osteoblastic cell line, which has several biological characteristics such as alkaline phosphatase activity, type I collagen synthesis, and matrix calcification research . Prolife3.1.3in vivo.The femur model is usually used for evaluating the dental implant materials, for the enclosed space facilitates the control of variable . There wHematoxylin and eosin (HE) staining , and van3.2The traditional treatment of peri-implantitis includes repeated complete debridement, replacement of dental implants, and intravenous antibiotic administration, the efficacies of which are often limited . Local ain vitro. As shown in S. aureus as well as C. albicans concentrations decreased. As shown in Once the biocompatibility of the PD was proved, we further investigated its antibacterial characteristics 3.3S. aureus and C. albicans was used. After three weeks of implant insertion, the rats were sacrificed and the Ti rod and bone tissue were collected to perform the CFU assay. The bacteria around the Ti rod were tested for the adhesive biofilm while the bacteria in the bone tissue demonstrated the planktonic microorganism. With an increase in the mass fraction of PD, total microorganism levels decreased (p < 0.05). The total microorganism level around the Ti rod in the 0.25 mg/mL group and the 0.5 mg/mL group was reduced by 2-log and almost 5-log compared with that in the PBS group, respectively  was found in 1993, and was thought to be a dentin-specific protein at first [dmp1 is overexpressed [dmp1 gene in the early stage (at 4 days and 7 days) and increased the expression level of dmp1 and alp gene in the middle stage (at 14 days). PD increased the expression levels of dmp1 and OPN in the early stage and increased the levels of alp in the middle stage. This demonstrates that PD has the potential to promote osteogenic differentiation of MC3T3-E1 cells.ALP, which is associated with the formation of hydroxyapatite crystals in osteoblasts, is a representative protein product of the osteoblast activity and is a good indicator of osteoblast differentiation. ALP activity is the most widely recognized biochemical marker for detecting osteoblastic activity ,73. Osteted gene . Dentin at first . Howeverxpressed . Accordi3.5X-ray evaluation was showed in Furthermore, severe pus formation was observed in the PBS group. Whereas, in the PD groups, the presence of less pus formation indicated that the clinical signs of pyogenic infections were greatly reduced. With an increase in the mass fraction of PD, the gross score significantly decreased (p < 0.05, We conducted qualitative and quantitative analysis of the bone tissue around the Ti rod subsequently. The analyses conducted at 3 weeks and 6 weeks post-surgery are showed in The morphological changes were observed by HE , and vanin vivo study. Hence, this is a double benefits material with great promise to prevent peri-implantitis during the two stages of implantation. Nonetheless, the fabrication process of PD still needs to be optimized. Because the PD was applied like a local drug, coating the PD on titanium are beneficial to the osteointegration due to the PAMAM retained on the implant surface after the DMADDM released. What's more, the two-staged double benefits dental implant need to be tested in larger animals.After three weeks, the rats in the PD groups showed less inflammation and bone destruction. Furthermore, compared with the rats in the PBS group, the rats in the PD groups showed reduced pus formation, lower radiographic and gross pathology scores. According to the data above, the use of MAO + PD is a successfully application for treating peri-implantitis. In the high-risk early stage, PD showed strong antibacterial and antifungal effects as well as low drug resistance, which was largely because of DMADDM. Besides, the cytotoxicity of PD was decreased because of the loading and slow releasing of PAMAM, therefore, allowing the new bone formation and osseointegration to happen. Moreover, in this study, PAMAM itself had the potential to promote osseointegration, which may be because of its ability to act as a scaffold for hard tissue regeneration. In the second stage, MAO has the potential to promote osteogenesis while maintaining a certain antibacterial effect; this was confirmed in the 4in vivo. Hence, PD + MAO is a kind of two-stage double benefits strategy of infection prevention and osteogenesis promotion. The modified dental implant may have a potential application in future clinical practice.This study demonstrated a PD + MAO dental implant with two-stage double benefits. In the high-risk first stage, PD enables to decrease the adherence and development of microorganisms by releasing DMADDM. Then, PD as well as MAO can promote osteogenesis and osseointegration. Furthermore, the PD + MAO strategy was proved to have great anti-infection and osteoconductive characteristics in a peri-implantitis rat model Xiaoyu Huang: Conceptualization, Methodology, Investigation, Writing – original draft. Yang Ge: Conceptualization, Methodology, Investigation, Writing – original draft. Bina Yang: Conceptualization, Methodology, Investigation, Writing – original draft. Qi Han: Investigation. Wen Zhou: Investigation. Jingou Liang: Investigation. Mingyun Li: Writing – review & editing. Xian Peng: Writing – review & editing. Biao Ren: Writing – review & editing. Bangcheng Yang: Writing – review & editing. Michael D. Weir: Investigation. Qiang Guo: Writing – review & editing. Haohao Wang: Investigation. Xinxuan Zhou: Investigation. Xugang Lu: Investigation. Thomas W. Oates: Writing – review & editing. Hockin H.K. Xu: Conceptualization, Writing – review & editing, Project administration. Dongmei Deng: Conceptualization, Writing – review & editing, Project administration. Xuedong Zhou: Conceptualization, Writing – review & editing, Project administration, Funding acquisition. Lei Cheng: Conceptualization, Writing – review & editing, Project administration, Funding acquisition.The authors declare no conflict of interest.


Escherichia coli gene deletion mutants and transcriptome sequencing (RNA-seq) data from 71 mutants to probe the extent of transcriptional and post-transcriptional effects of gene deletions in operons. Transcriptional effects were common on genes located downstream of the deletion and were consistent across all operon members, with nearly 40% of operons showing greater than 2-fold up- or downregulation. Surprisingly, we observed an additional post-transcriptional effect that leads to the downregulation of the gene located directly downstream of the targeted gene. This effect was correlated with their intergenic distance, despite the ribosome binding site of the gene downstream remaining intact during library construction. Overall, the data presented can guide future library construction and highlight the importance of follow-up experiments for assessing direct effects of single-gene deletions in operons.Single-gene deletions can affect the expression levels of other genes in the same operon in bacterial genomes. Here, we used proteomics for 133 IMPORTANCE Single-gene deletion libraries have allowed genome-wide characterization of gene function and interactions. While each mutant intends to disrupt the function of a single gene, it can unintentionally target other genes, such as those located in the same operon as the deletion. The extent to which such polar effects occur in deletion libraries has not been assessed. In this work, we use proteomics and transcriptomics data to show that transcript level changes lead to nearly 40% of deletions in operons affecting the protein levels of genes located downstream by at least 2-fold. Furthermore, we observed a post-transcriptional effect on the gene located directly downstream of the deletion. These results can guide the design of future gene deletion libraries and emphasizes the importance of follow-up work when linking genotypes to phenotypes. Download FIG S1, TIF file, 0.7 MB.Protein and transcript levels of genes located in the same operon as a deleted gene. Each line represents one deletion mutant and the protein and RNA expression levels of other operon members . Protein (76 deletion mutants): n−4 = 5, n−3 = 9, n−2 = 18, n−1 = 46, n1 = 57, n2 = 32, n3 = 19, n4 = 10. RNA (26 deletion mutants)  : n−2 = 7Copyright © 2021 Mateus et al.2021Mateus et al.https://creativecommons.org/licenses/by/4.0/Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the 10.1128/mSystems.00813-21.3FIG S3n−4 = 31, n−3 = 31, n−2 = 32, n−1 = 32, n1 = 24, n2 = 28, n3 = 29, n4 = 27. (b) As in panel a but for RNA levels. n−4 = 31, n−3 = 29, n−2 = 30, n−1 = 34, n1 = 29, n2 = 27, n3 = 31, n4 = 27. Download FIG S3, TIF file, 0.6 MB.Levels of genes located in different operons as a deleted gene. (a) Protein levels are shown as in Copyright © 2021 Mateus et al.2021Mateus et al.https://creativecommons.org/licenses/by/4.0/Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the 10.1128/mSystems.00813-21.4FIG S4ahpC, Δfis, ΔiscS, ΔiscU, ΔsufB, and ΔsurA mutants. r = 0.63, P = 0.011, n = 15. Download FIG S4, TIF file, 0.3 MB.Comparison of protein and transcript levels for overlapping mutants between this study and Parker et al. . ProteinCopyright © 2021 Mateus et al.2021Mateus et al.https://creativecommons.org/licenses/by/4.0/Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the 10.1128/mSystems.00813-21.8TABLE S1Table S1, XLSX file, 18.9 MB.Proteome-wide protein levels of mutants analyzed in this study and in the study by Mateus et al. . DownloaCopyright © 2021 Mateus et al.2021Mateus et al.https://creativecommons.org/licenses/by/4.0/Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the r = −0.31, P = 0.071, n = 34)  see , support 34) see ; so, in 10.1128/mSystems.00813-21.5FIG S5n1 = 16, n2 = 16, n3 = 7, n4 = 4. (b) (Top) Schematic representation of excision of kanamycin resistance cassette, leading to a genomic scar that codes for a small in-frame peptide composed of the first and last seven amino acids of the original gene, with some additional residues from FRT site—total length 34 amino acids  Subsets from Copyright © 2021 Mateus et al.2021Mateus et al.https://creativecommons.org/licenses/by/4.0/Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the 10.1128/mSystems.00813-21.6FIG S6x axis) were calculated for genes downstream of the deletion (excluding the gene directly downstream), and median operon protein abundance (y axis) was calculated for the same genes based on the average top-3 parameter  across all mutants in the study by Mateus et al.  but not at the mRNA level (P = 0.93 Wilcoxon rank sum test) (P = 9.1 × 10−5 Wilcoxon rank sum test) remaining after normalizing out the general effect on expression for upstream and downstream genes  . This deam genes . This efam genes .Thus, this strongly suggests that the overall downregulation of the gene located directly downstream of a deletion is post-transcriptional. Since the Keio mutants have been designed to avoid obvious sources of translational polarity , we decided to look further into the source of this effect.fruB and ΔpabC) and one that did not (ΔyecN), and built different versions of the deletion in which the FRT sites were removed from either or both sides of the resistance cassette . Download FIG S7, TIF file, 0.9 MB.Effects of FRT sequence on the levels of the gene directly downstream of a gene deletion. (a) Schematic representation of mutants built for evaluation of their effects. (b) Position of FRT sequence does not affect the levels of the gene directly downstream of a gene deletion. Data are shown as in Copyright © 2021 Mateus et al.2021Mateus et al.https://creativecommons.org/licenses/by/4.0/Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the rS = 0.62, P = 2.5 × 10−4, n = 30)   .Next, we observed a trend between how much the gene directly downstream of the deleted gene is downregulated  and their intergenic distance—with genes that were at a closer distance showing stronger downregulation ( n = 30) . Thus, wd ΔrfbB) . Althoug n = 55) , left. Eonstruct , right aOverall, these results suggest that translation initiation problems, a lack of translational coupling, or both, depending on the deletion, partially explain why the levels of the gene downstream of a deletion are affected. Furthermore, it seems that translational coupling is affected by the size of the gene being translated, with 34 residues not being enough for most genes (as seen above for a larger number of mutants) , but incybaB deletion mutant is sensitive to UV, not because cells lack YbaB but because of the low levels of the adjacently encoded RecR , show widespread polar effects. Nearly 40% of the gene deletions profiled here showed >2-fold up- or downregulation of transcripts levels of genes located downstream of a deleted gene. Upregulation is likely caused by the presence of the promoter of the resistance cassette, which boosts expression of low-expressed genes. In contrast, the downregulation might be caused by disturbances to intraoperon regulatory sequences or mRNA stability of highly expressed genes. We also showed a post-transcriptional effect on the gene that is located directly downstream of the deleted gene, which generally showed lower protein levels than the rest of genes of the operon. This effect was correlated with the intergenic distance between the deleted gene and the downstream gene. Our follow-up work suggests that problems with translation initiation or lack of translational coupling lead to this effect, but further studies will be required to properly understand the underlying mechanism. Future library design could be improved by increasing the C-terminal scar of the deleted gene. When using existing libraries, it is important to be aware that any such polar effects can lead to the misinterpretation of phenotype-genotype associations, and complementation experiments  are important follow-ups to distinguish direct from indirect effects. As an example, we have recently found that the ded RecR . More brtrometry , 27, 28.E. coli BW25113 strain background. When available, mutants were used directly from the Keio collection using two independent clones  (All mutants used in this study have been made in the licates)  (see Tablicates)  using th10.1128/mSystems.00813-21.9TABLE S2Table S2, XLSX file, 0.1 MB.Details of mutants used in this study. Download Copyright © 2021 Mateus et al.2021Mateus et al.https://creativecommons.org/licenses/by/4.0/Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the 578) of ∼0.5, washed with phosphate-buffered saline (PBS), and lysed by the addition of 2% SDS followed by an incubation at 95°C for 10 min.Protein level measurements that did not originate from Mateus et al.  were perE. coli (strain K-12) UniProt FASTA (proteome identifier [ID] UP000000625) using isobarQuant . Then, for each gene, we retrieved the chromosome positions from the annotations in NCBI GenBank accession number NC_000913.2, summed up all counts within those positions for each mutant, and divided this by the gene length. We removed all cases in which this count value normalized by gene length was <1. For each gene in each mutant, we then calculated the relative transcript levels to that in the wild type by calculating the ratio between the count value normalized by gene length by the median count value normalized by gene length across all mutants.We downloaded the WIG files from the data set https://ecocyc.org/) (2 fold change (protein or transcript) for all genes in the same operon located upstream of the gene of interest and subtracted this value from the log2 fold change for every gene located upstream of the gene of interest. We repeated the same procedure for genes located downstream of the gene of interest.We retrieved the operon structure from Ecocyc v21.1 (yc.org/) . Then, fGSE129161).The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the data set identifier PXD023945. We further retrieved data from the PRIDE repository with the accession number PXD016589. RNA-seq data were collected from the Gene Expression Omnibus (accession number PXD023945 and PXD016589) and the log2 fold change of mutant versus wild type (https://github.com/andrenmateus/gene_deletion_effects_on_operons.The code to process raw mass spectrometry data (available at PRIDE partner repository with the data set identifiers ild type  is avail10.1128/mSystems.00813-21.10DATA SET S1Data Set S1, XLSX file, 0.1 MB.Raw data used for generating figures and supplementary figures. Download Copyright © 2021 Mateus et al.2021Mateus et al.https://creativecommons.org/licenses/by/4.0/Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the


The purpose of this study was to evaluate the ocular biometric parameters in adult cataract patients from China and create an anterior chamber depth (ACD) regression model.The ocular biometric records of 28,709 right eyes of cataract surgery candidates who were treated at Aier Eye Hospitals in nine cities from 2018 to 2019 were retrospectively analyzed. All measurements were taken with IOLMaster 700. We included patients who were at least 40 years old and were diagnosed with cataract.P < 0.01). In the multivariate regression analysis of ACD, which included LT, AL, WTW, sex, Km, CCT, and age, there was a reasonable prediction with adjusted R2 = 0.641.The mean age of the patients was 68.6 ± 11.0 years. The mean values recorded were as follows: axial length (AL), 24.17 ± 2.47 mm; mean keratometry (Km) value, 44.26 ± 1.70 D; corneal astigmatism (CA), 1.06 ± 0.96 D; ACD, 3.02 ± 0.45 mm; lens thickness (LT), 4.52 ± 0.45 mm; central corneal thickness (CCT), 0.534 ± 0.04 mm; and white to white (WTW) corneal diameter, 11.64 ± 0.46 mm. ACD correlated positively with AL  and WTW (0.300), but negatively with LT (-0.660) and age (-0.285) (all Cataract patients with longer AL and wider WTW have deeper ACD. With increasing age and lens thickening ACD becomes shallower. Based on the standardized coefficients of ACD multivariate regression analysis from the study, LT is the main factor that affects ACD, and is followed by AL. Modern cataract surgery has transformed from a rehabilitative procedure to a refractive operation. This can be attributed to the development of measuring devices, lens calculation formulas, surgical technology, and intraocular lenses. Accurate prediction of refractive outcomes after cataract surgery is crucial. When biometry is based on optical coherence interferometry (Zeiss IOLMaster), the error from anterior chamber depth (ACD) prediction amounts to approximately 42 % of the total refractive prediction error . Olsen . –12. 8–12 surgery , 14 and  surgery , 16 haveP = 0.005). This is similar to the results of Jivrajka’s report (mean: 75 years versus 73 years), which considered that women had a longer life expectancy than men [In present study, more female patients scheduled cataract surgery than male patients (59.1 % versus 40.9 %). This finding is almost identical to those of previous published studies   and shallower ACD  than male patients. This echoes the findings of other studies of cataract patients in different countries [In the present study, female patients presented for surgery with a shorter average AL   was 6.4 %, which is similar to that of other studies , 28; theThe mean ACD in the present study was 3.02 ± 0.45 mm. From the matrix of Spearman correlation of biometric parameters (Table R2 = 0.641 means that 64.1 % of the variance could be explained by this model. A study of an elderly Chinese population showed that approximately one in five people aged 50 years and over developed some form of angle closure over a 10-year period, and reported best cut-off values of 2.60, 4.72, and 22.92 mm for ACD, LT, and AL, respectively, in predicting incident PAC [Based on the standardized coefficients of ACD multivariate regression analysis, LT is the main factor that affects ACD, and is followed by AL Table . Adjustedent PAC . In the This study has certain limitations. First, the data from the nine hospitals do not completely represent the ocular parameters of the overall population in China. Second, we need to collect more information about anthropometric characteristics, education, occupation, income, and the type of intraocular lens, and evaluate the correlations among them. We will conduct more research to improve on the findings of the analyses.In summary, we collated data on the ocular biometric parameters of cataract surgery candidates in China, compared them, analyzed their correlations, and created an ACD regression model. This study provided reference values for AL, Km, CA, ACD, LT, CCT and WTW by using IOLMaster 700 for adult cataract patients in China. Cataract patients with longer AL and wider WTW have deeper ACD. With increasing age and lens thickening ACD becomes shallower. Based on the standardized coefficients of ACD multivariate regression analysis from the study, LT is the main factor that affects ACD, and is followed by AL.


Barrett's esophagus (BE) is a well‐established risk factor for esophageal adenocarcinoma. Our objective was to investigate the effectiveness of lifestyle interventions on BE risk.We searched PubMed, Embase, and Web of Science up to 30 September 2020. The summary relative risks (RRs) and 95% confidence intervals (CIs) for the highest versus lowest categories of exposure were assessed. Analyses of subgroup, dose–response, sensitivity, and publication bias were conducted.p < 0.05). There was no publication bias for smoking and alcohol. Although the analysis suggested significant evidence of publication bias for BMI, sensitivity analysis showed that the changes in the recalculated RRs were not significant.Sixty‐two studies were included that involved more than 250,157 participants and 22,608 cases. Seven lifestyle factors were investigated: smoking, alcohol, body mass index (BMI), physical activity, sleep time, medication, and diet. We observed statistically significant increased BE risks for smoking , alcohol intake , body fatness , less sleep time , and proton pump inhibitors use . Reduced risks of BE were found for aspirin  and the intake of vitamin C , folate , and fiber . The quality of most included studies was high and the subgroup analysis according to the quality score showed significant results (The large meta‐analysis revealed that lifestyle modifications could reduce the risks of BE and, consequently, esophageal adenocarcinoma. We conducted a large meta‐analysis based on sufficient evidence and a quantitative synthesis of the eligible data published up to 30 September 2020, to investigate the effectiveness of lifestyle interventions on Barrett's esophagus risk. In this meta‐analysis of 62 studies that involved more than 250,000 participants and 22,000 cases, we found that smoking, alcohol intake, high levels of body fatness, and less sleep time are associated with Barrett's esophagus risk. There are statistically significant reduced risks of Barrett's esophagus with aspirin use and vitamin C intake, folate, and dietary fiber. In a word, these findings strengthen our understanding of the potential mechanisms of Barrett's esophagus development and highlight awareness that lifestyle intervention could reduce the risks of Barrett's esophagus and, consequently, esophageal adenocarcinoma. Details of the search terms  were: “lifestyle,” “risk(s),” “diet/dietary,” “food(s),” “smoking/smoker/tobacco/cigarette(s),” “drink/drinking/alcohol/ethanol/alcoholic/beverage(s)/wine/beer/spirits/liquor,” “fatness/obesity/obese/obeseness/adiposity/overweight/weight/body mass index/BMI/waist hip ratio/waist circumference/hip circumference,” “physical activity/exercise,” “sleep/nocturnal,” “medicine/medical/medication(s),” “nonsteroidal anti‐inflammatory drug(s)/NSAID(s)/ibuprofen/diclofenac/naproxen/indomethacin/mefenamic acid/piroxicam/ketoprofen/etodolac/meloxicam/rofecoxib/flurbiprofen/phenylbutazone/aspirin,” “proton pump inhibitor(s)/PPI(s)/omeprazole/pantoprazole/esomeprazole/lansoprazole/dexlansoprazole,” “statin(s)/hydroxymethylglutaryl‐CoA reductase inhibitor/simvastatin/lovastatin,” “nutrition/nutrient,” “vitamin(s),” “folate/folic acid,” “fiber(s)/fibre(s),” “meat(s)/fish/poultry/chicken/turkey/duck,” and “selenium” in combination with “Barrett's/Barrett,” “esophagus/oesophagus/esophageal/oesophageal/neoplasia/neoplasm/neoplasms,” The two sets of keywords were combined individually, and the eligibility criteria were independently judged by two reviewers (ZZ and ZY).2.3The study quality of cohort studies and case–control studies was assessed using the Newcastle–Ottawa Scale (NOS).A data extraction sheet was generated for each study. Detailed information included the first author, publication year, country, study type, study period, study population, assessment method, type of exposure measured, exposure categories, adjusted RR (95% CI), adjusted variables, and quality score.2.4SPSS 22.0  was used to collect and extract data. RevMan5.3  software was used for the synthesis and analysis of data based on relative risks (RRs) and 95% confidence intervals (95% CIs).We conducted this meta‐analysis for the risk of BE and smoking, BMI, physical activity, sleep duration, medications, and dietary factors. Medications included aspirin, NSAIDs, PPIs, and statins. Dietary factors included alcohol, vitamin C, folate, selenium, total meat, and white meat. Stratified analysis was not performed for alcohol. Beer, wine, and spirits were included. Fruits, vegetables, fat, red meat, and processed meat were excluded because we have previously analyzed these issues.2I statistics p < 0.1 was considered representative of significant heterogeneity). Gastroesophageal reflux disease (GERD) is a well‐established risk factor for the development of BE.Heterogeneity among studies was detected using p < 0.1 was considered significant publication bias).Publication bias was assessed using funnel plots and Egger's test   Figure . The cha3.2.3p = 0.72, 2I = 0%). The dose–response analysis of the number of cigarettes/day was not conducted due to the limited studies.Four studies were included in the analysis for the highest to lowest number of cigarettes/day Figure , and a f3.2.4Pnonlinearity < 0.01, Figure We conducted a dose–response analysis to further explore the association between pack‐years of smoking and BE risk. Six studies were included, and the results of 1.10 (1.05–1.14) indicated that BE risk increases by 10% for each 10‐year increment. We further checked for nonlinearity of the dose–response association, and the evidence suggested that the best‐fitting model was a nonlinear model , but subgroup analyses (Table There was significant heterogeneity (3.2.6p < 0.1), but the funnel plot provided a visible result of no publication bias observed in Figure A funnel plot, Begg's test, and Egger's test were used to explore the publication bias. Indeed, Egger's test indicated evidence of publication bias   and subgroup analyses also suggested that the positive association was stable by all of the confounders (Table There was no significant heterogeneity (rs Table .3.3.3p = 0.34) suggested no significant evidence of publication bias. Additionally, the sensitivity analysis also showed that the changes in the recalculated RRs were not significant, with a range from 1.20 (1.09–1.33) when excluding Avidan 2001 (27.2%) to 1.26 (1.15–1.37) when excluding Park 2009 (5.9%).The funnel plot Figure  and Egge3.43.4.1Twenty‐two studies that involved 211,607 participants were included, and a random‐effects model yielded a positive association  Figure .3.4.22 increase in BMI. We further checked for nonlinearity of the dose–response association, and the evidence suggested that the best‐fitting model was a nonlinear model  indicated that the BE risk increases by 8% for each 5 kg/m3.4.3p < 0.01, 2I = 63%), but subgroup analyses (Table There was significant heterogeneity (es Table  suggeste3.4.4p < 0.1) suggested significant evidence of publication bias. However, sensitivity analysis showed that the changes in the recalculated RRs were not significant, with a range from 1.06 (1.02–1.10) when excluding Edelstein 2007 (1.4%) to 1.18 (1.08–1.29) when excluding Thrift 2014 (29.8%).Indeed, the funnel plot Figure  and EggeFive studies investigated BE risk and aspects of waist‐to‐hip ratio (WHR), waist circumference, hip circumference, waist‐to‐thigh ratio, and visceral adiposity. These investigations suggested that high WHR,3.53.5.1p = 0.77, 2I = 0%)  without heterogeneity  Figure . Additio3.5.2p = 0.53, 2I = 0%)  without heterogeneity  Figure . The cha3.63.6.1p = 0.04, 2I = 52%)  with heterogeneity  Figure . Additio3.6.2Six studies were included that involved 3,742 participants. A fixed‐effects model yielded an inversed association  Figure . The cha3.6.3Seven studies were included that involved 14,908 participants. A significant increased BE risk was observed  Figure . The sen3.6.4Four studies were included that involved 4,845 participants. A significant reduced BE risk was observed  Figure . The sen3.7Significant inverse associations were observed between BE risks and the highest versus lowest intakes of vitamin C  may be an independent risk factor for BE development. Nevertheless, long‐term smoking cessation may diminish this risk,http://monographs.iarc.fr/ENG/Classification/ClassificationsGroupOrder.pdf) and the World Cancer Research Fund International  have classified alcohol as a Group 1 carcinogen for esophageal cancer. Thus, a decreased intake of alcohol is advisable to reduce the risk of BE and esophageal adenocarcinoma.A significant 23% increased risk was observed for highest versus lowest alcohol intake and BE risk, which was supported by the results of detailed subgroup analyses. Moreover, the International Agency for Research on Cancer . Thus, keeping the weight as low as possible within the healthy range is helpful to reduce the risks of BE and esophageal adenocarcinoma.Although Qumseya et alPhysical activity is often considered an inverse factor for esophageal cancer.H pylori infection in combination with antibiotics, and positive H pylori infection is associated with a reduced risk of BE.It is surprising that aspirin use was protective against BE with a 30% lower risk, but that nonaspirin NSAID use was not. The results were consistent with the results of the largest studyA 41% decreased risk was observed for the highest versus lowest intake of vitamin C and BE risk. Protective associations were also observed with the intake of folate and dietary fiber. Analyses for the highest versus lowest intakes of total meat, white meat, and selenium yielded nonsignificant risks of BE. Systematic analyses could not be conducted for other common dietary factors, such as other vitamins, calcium, tea, and coffee, because of the limited studies, and further studies are required to further validate our findings and to reveal these uncertain conclusions.Our study has several strengths. The first strength was that our systematic analysis was based on the main modifiable lifestyle factors, the substantial sample size and the quantitative synthesis of the eligible data, which provided sufficient robust and reliable evidence and increased the statistical power of our findings. Second, we performed detailed subgroup analyses and dose–response analyses to further detect the associations rather than simply conducting categorical comparisons. These independent analyses provided accurate evaluations and strengthened the conclusion. Third, we broadly and systematically searched three large databases to identify studies published from inception through 30 September 2020, and the reference lists of the included studies were also searched manually to identify additional literature. Two reviewers selected the studies and extracted the data independently and in duplicate, which increased the validity of our analyses. Last, 62 included studies were identified from 16 countries or regions in the Americas, Europe, Asia, and Australia, which increased the generalizability of our results.Hpylori infection, and hiatal hernia) could still not be obtained in several studies. Thus, our results should be considered carefully due to possible confounding. Third, the ranges of the highest to lowest category varied in the included studies, which influenced the accuracy of the results to some extent, and we cannot thoroughly exclude the potential bias. Nevertheless, to reduce the bias to a large extent, the pooled results for the highest compared with lowest category were adopted and pooled, and the results were further verified by dose–response analyses, which yielded results similar to the original analyses. Finally, the language of the included studies was limited to English, which may lead to potential selection bias.However, the limitations of the present meta‐analysis should be taken into consideration. First, the diagnostic criteria of BE may vary among the included studies.5This large systematic analysis demonstrated that smoking, alcohol intake, high BMI, and less sleep time are associated with BE risk. There are statistically significant reduced risks of BE with aspirin use and the intake of vitamin C, folate, and dietary fiber. Our findings strengthen our understanding of the potential mechanisms of BE development and highlight an awareness that lifestyle interventions may reduce the risks of BE and, consequently, esophageal adenocarcinoma.The authors declare no potential conflict of interest.Zhanwei Zhao and Zifang Yin wrote the main manuscript and participated in the study design and the data analysis. Chaojun Zhang completed the design of the work. All the authors have reviewed the manuscript text. ZZ and ZY contributed equally to this work.Fig S1‐S4Click here for additional data file.


It was found that in contrast with the control group, the MAP in the experimental group at T2, T3, and T4 periods was lower, while it was higher at T5 period and its HR at T2, T3, T4, and T5 periods was higher (P < 0.05); the PAED and FLACC scores were lower (P < 0.05), and the incidence of ADE (10.53%) was lower than that in the control group (31.58%) (P < 0.05). However, SpO2 at different periods showed no obvious differences between the two groups (P > 0.05). In conclusion, the restoration algorithm-based ultrasound images had high quality, and they demonstrated good application value in evaluating the effect of Dex to prevent neurological disorders in patients anesthetized by sevoflurane.In this article, dexmedetomidine (Dex) was used to prevent neurological disorders in patients anesthetized with sevoflurane and the effect was analyzed using ultrasound images based on the restoration algorithm of the linear system model. Children injected with Dex were in the experimental group, while children injected with normal saline were in the control group. The mean arterial pressure (MAP), arterial oxygen saturation (SpO Ultrasonic imaging is an acoustic technology based on ultrasonic pulse reflection, which uses the ultrasonic beam to generate reflected signals at the interface of tissues and then obtains the images of internal organs . Althougα2 receptor agonist, is widely used in sedation, analgesia, and antianxiety. It can act on α2 receptors to inhibit the release of norepinephrine, thereby achieving sedation, analgesia, and antianxiety [μg/kg Dex can reduce the incidence of emergency agitation in children and maintain good hemodynamics of children in operation [Inhalation anesthesia has been used clinically since a hundred years. Inhalation anesthesia is usually used for the induction and maintenance of anesthesia in pediatric operations. Sevoflurane is an anesthetic drug with relatively stable induction. Because of its fast induction of anesthesia, no irritating smell, and low blood distribution coefficient, it is more suitable for pediatric anesthesia. At present, sevoflurane is widely used clinically , 5. Relaianxiety . Relevanperation .To sum up, to obtain high-quality ultrasound images and better assist doctors, it is necessary to improve the original ultrasound images through ultrasonic image restoration technology. In the study, the linear model of ultrasound images and the working principle of the Field II ultrasound simulation platform were adopted for analysis. Then, the Field II method was used to estimate the PSF. Finally, the ultrasound image was restored by the deconvolution restoration method. Sevoflurane inhibits the respiratory function of children, while Dex drugs not only can reduce the pain and stress response of children but also have no significant inhibition on the respiratory function of children. Therefore, based on the ultrasonic image restoration, the effect of Dex was investigated on preventing sevoflurane emergency agitation in children.In this study, 76 children undergoing tonsillectomy were selected as the research subjects, who were admitted to the hospital from February 2018 to January 2019. They were between 3 and 10 years old and rolled into a control group (38 cases) and an experimental group (38 cases) in random. In the control group, there are 25 males and 13 females, with an average age of (6.13 ± 2.78) years. In the experimental group, there were 23 males and 15 females, with an average age of (6.35 ± 2.89) years. The study has been approved by the medical ethics committee of the hospital, and the patients and their family members were informed and signed consent forms.Inclusion criteria were as follows: (i) patients older than 2 years; (ii) patients classified as I and II by ASA; (iii) patients who have not received surgical treatment; (iv) patients younger than 11 years; (v) patients undergoing normal examination with consciousness.Exclusion criteria were as follows: (i) patients with mental illness; (ii) patients with liver and kidney dysfunction; (iii) patients allergic to sedative drugs; (iv) patients with incomplete clinical data; (v) patients who withdrew from the experiment due to their own reasons.x and y represent the horizontal and vertical coordinates, respectively, ∗ represents a discrete two-dimensional linear convolution operator, G  represents the received ultrasonic radio-frequency (RF) data, K  is a two-dimensional point-diffusion function, P  is the tissue-reflectivity image, and U  represents additive noise and is assumed to be white noise. G  to P .The PSF and the tissue reflectivity convolve and accumulate the noise. The restoration process of ultrasound images can be described with a linear system model. The ultrasound image restoration can be expressed as follows:PSF estimation is the key step in the ultrasound image restoration. In the study, the Field II method was adopted to study the two-dimensional PSF, which was based on the Field II ultrasonic simulation platform. It was assumed that the PSF is constant in the lateral direction and variable in the axial direction. Therefore, the received RF echo data can be segmented axially. The PSF corresponding to each segment can be estimated by the Field II method. Then, each segment accepted deconvolution restoration, and all segments after the deconvolution and restoration were put together to form the final image. The unconstrained inverse filtering deconvolution restoration method is adopted.G, P, U, and K are the Fourier transform of the observed image G, the tissue-reflectivity image P, noise U, and point-diffusion function K. According to equation  can be obtained by performing Fourier Transform on P. The1/K in equation  is just the reverse of the imaging system K. Therefore, the process is called the inverse filtering restoration method. The following equation is obtained by performing inverse Fourier transform on equation . Then, tracheal intubation was performed, and an anesthesia machine  was employed to control the breathing mechanically. Patients in the experimental group were given 1 μg/kg.h Dex  by continuous intravenous pump injection, and patients in the control group were given the same amount of normal saline. After the operation, the sevoflurane inhalation and Dex pumping were stopped.All patients underwent fasting for 4 hours and were forbidden to drink for 2 hours before the operation, and no drugs were taken. Before the operation, the patients' intravenous trocar should be indwelled, and oxygen and aspiration devices, first aid drugs, and equipment should be prepared. In the operating room, the patients' routine blood pressure, blood oxygen, etc. were checked. Both groups of patients were treated with sevoflurane  according to the standard. After the patients lost consciousness, an intravenous channel was established for intravenous injection of 0.1 mg/kg of vecuronium  and 2 2, and HR in the two groups were detected with Philips VE24 (USA) before the injection (T1), at 5 min (T2), 10 min (T3), and 20 min (T4) after the injection, and when the patient came to himself (T5). After the drug was stopped, the children were sent to the resuscitation room and awakened 10 minutes later. The nursing staff used a single-blind method to evaluate the child's PAED and FLACC scores. The scoring criteria of PAED were as follows: no restlessness scored less than 5 points; mild restlessness scored 5–10 points; severe restlessness scored 10–15 points. FLACC is a scale used to assess the degree of pain in infants and children who cannot confirm the pain/quantify the degree of pain. Its takes account of expressions, leg movement, body position, crying, and comfort. The lowest score is 0, which means there is no pain. Mild pain scores 1–4 points, obvious pain scores 5–8 points, and severe pain scores 9–10 points. The occurrence of ADE and its incidence after surgery were recorded.The MAP, SpOx ± s) illustrated how to calculate the measurement data, and the count data were expressed as a percentage. MPA, SpO2, HR, and PAED scores during anesthesia between the two groups were compared by the paired t-test. Analysis of variance was adopted for comparison among groups. The difference was statistically significant with P < 0.05.The data were processed by SPSS19.0, the mean ± standard deviation (‾P > 0.05)  .P < 0.05), indicating the PSF obtained by the generalized homomorphic filtering method was closer to the PSF estimated by the Field II method than the PSF obtained by the complex cepstrum method.As shown in Since the PSF estimated by the Field II method was closer to the real PSF, it was selected to estimate the PSF in this study. Then, unconstrained inverse filtering deconvolution was adopted to restore ultrasound images. As shown in P > 0.05), while in contrast with the control group, the MAP values of the experimental group at T2, T3, and T4 periods were obviously lower, but value at T5 period was obviously higher (P < 0.05).As shown in P < 0.05) , while the HRs of the experimental group at T2, T3, T4, and T5 periods were obviously higher (P < 0.05).The HRs of the control group at T1, T2, T3, T4, and T5 periods were 103.23 ± 17.35, 95.33 ± 16.68, 93.65 ± 17.47, 92.16 ± 16.91, and 106.74 ± 17.18 (times/min); those of the experimental group were 102.35 ± 18.58, 100.23 ± 17.85, 101.54 ± 18.12, 99.69 ± 18.24, and 126.86 ± 17.93 (times/min), respectively. As shown in 2 rates of the control group at T1, T2, T3, T4, and T5 periods were 99.31 ± 0.62, 98.23 ± 0.69, 98.67 ± 0.59, 99.30 ± 0.71, and 98.89 ± 1.02 (%), respectively; those of the experimental group were 99.47 ± 0.45, 99.25 ± 0.37, 98.69 ± 0.51, 98.58 ± 1.16, and 99.57 ± 0.34 (%), respectively. As shown in 2 between the two groups at T1, T2, T3, T4, and T5 periods (P > 0.05).As shown in P > 0.05); while the PAED scores of the experimental group at T2, T3, and T4 periods were obviously lower than those of the control group (P < 0.05).As shown in P > 0.05); however, the FLACC scores of the experimental group at T2, T3, T4, and T5 periods were obviously lower (P < 0.05).As shown in P < 0.05).As shown in α2 receptor agonist. It is mainly used for compound anesthesia, ICU sedation, preoperative medication, imaging-assisted examination, postoperative agitation prevention, etc. [At present, ultrasound imaging technology is widely used in medical diagnosis and detection and has become the most commonly used method . Howeveron, etc. . The app2, HR, and PAED and FLACC scores of the two groups at different time periods were detected; there was no obvious difference in MAP between the two groups at T1 period (P > 0.05), while the MAP of the experimental group at T2, T3, and T4 periods was lower and that at T5 was higher (P < 0.05). The HRs of the experimental group were 102.35 ± 18.58, 100.23 ± 17.85, 101.54 ± 18.12, 99.69 ± 18.24, and 126.86 ± 17.93 (times/min), respectively, and were higher than those of the control group at T2, T3, T4, and T5 periods (P < 0.05). There was no obvious difference in SpO2 between the two groups at different time periods (P > 0.05). The PAED and FLACC scores of the experimental group were significantly lower than those of the control group at T2, T3, and T4 periods (P < 0.05). The incidence of ADE in the experimental group was significantly lower than that in the control group (P < 0.05). This was similar to the results of Di et al. [In the study, the PSF was estimated by the Field II method and the ultrasound image was restored by the deconvolution restoration method. Then, the effect of Dex was investigated on preventing sevoflurane emergency agitation in children. It was found that the PSF estimated by the Field II method was closer to the real PSF. Furthermore, the PSF obtained by the generalized homomorphic filtering method was closer to the PSF estimated by the Field II method than the PSF obtained by the complex cepstrum method. It was aligned with the research results of Jemec et al. . High-rei et al. , suggestIn the study, the linear model of ultrasound images and the working principle of the Field II ultrasound simulation platform were adopted for analysis. The ultrasound images were restored by the deconvolution restoration method to obtain the ones with higher quality. On the basis of ultrasound image restoration, the effect of Dex was investigated on preventing sevoflurane emergency agitation in children. It turned out that Dex can effectively reduce the incidence of emergency agitation and the adverse conditions after the operation. However, the sample size is small, which may cause deviations in the results. In the follow-up studies, the number of patients will be increased to further explore the effect of Dex on preventing sevoflurane emergency agitation in children. In summary, the results provide a theoretical basis for the study of ultrasound image restoration algorithms and Dex's prevention of neurological disorders in patients undergoing sevoflurane anesthesia .


However, untrained subjects could not distinguish the difference in the quality of the 2 sets of cabbage juices in the triangle test . Regardless of the distinguishing features of color, aroma, and taste, the subjects did not have any preference between the two different cabbage juices.To date, the impacts of agriphotovoltaic (APV) condition on the production yield of crop have been studied; however, the effect of APV production on the sensorial quality and consumer acceptability of the produce remains unexplored. Therefore, to address this knowledge gap, we cultivated “Winter Storm” cabbage under solar panels (20.16 kW) and in open field in 2020. The weight and diameter reduction rate of fresh cabbage grown under APV condition compared to open-field conditions were 9.7% and 1.2%, respectively. The levels of glucosinolates and their hydrolysis products were not significantly different in the fresh cabbage between the two conditions. The amount of volatile organic compounds, which may affect the perception of smell, were significantly higher in the cabbage juice prepared from the ones grown in open-field conditions than in the juice prepared from cabbages grown under APV conditions ( As environmental problems, such as global warming caused by fossil fuel use, have emerged, renewable sources of energy have become an effective alternative. Several countries have announced climate-friendly energy policies . South KAccording to the Japanese energy policy, crop yield under APV should not be less than 80% of those grown in open-field (OF) conditions to ensure food security . Thus, sRaised solar panel in APV can change the microclimate because of its structure above the crops. The shaded part, caused due to the blocking of the sunlight by the PV panels, have decreased soil temperature and lower light intensity reaching the plants ,12. In gBrassica vegetables, including cabbage (Brassica oleracea var. capitata), contain anticarcinogenic phytochemicals, especially glucosinolates and its hydrolysis products [B. oleracea var. italica). Changes in environmental factors including air temperature, soil temperature, and soil water content during growing season affect the content of glucosinolates, especially indolyl glucosinolates [products ,16. In cproducts ,18. Glucproducts . Chiu etproducts  suggesteinolates ,22,23. TAccording to a report by the Ministry of Agriculture, Food, and Rural Affairs on food and dining statistics in 2020, the market share of vegetable juices in South Korea was 0.15 billion dollars. People began drinking cabbage juice because it was easy to consume and effective in recovery from stomach ulcers . HoweverUntil now, research has been focused on the growth and yield of crops grown under APV condition or possible species selection ,28,29,302, with 3.3 m high columns (with a spacing of 4 m × 5 m). The specifications and layout of the PV panel were as follows: each panel had the length, width, and thickness of 1.68 m, 0.95 m, and 30 mm, respectively. The angle of panels was 35°. Four bi-facial panels were in four columns. The total power generation capacity of PV panels was 20.16 KW . The total area of the APV structure was 480 m20.16 KW .Microclimate data were collected using data loggers . We collected data on atmospheric and soil variables. Specifically, air temperature and humidity, solar radiation, and soil temperature and moisture were collected using ATMOS 14, PYR, and TEROS 11 sensors, respectively. ATMOS 14 and PYR sensors were installed 60 cm above the ground, and TEROS 11 sensors were installed 20 cm under the ground.B. oleracea var. capitata, cultivar: “Winter Storm”) was purchased from Asia Seed Co. Ltd. , and sown on 27 July 2020, in 105-cell seedling trays. The seedlings were transplanted into 30 cm raised bed, covered with black plastic mulching film at 35 × 35 cm intervals, in the farm of Chonnam National University on 6 September 2020. NovaTec-Suprem fertilizer  was applied at 100 g·m−2 to provide nutrients. Cabbages were harvested on 15 January 2021 to avoid extreme cold weather. In the OF and APV area, 5 cabbages were sampled and pooled per replication site . We weighed and measured the diameter of the cabbages using a scale and a ruler immediately after harvest. The diameter was measured at the longest axis of the width. A total of 5 fresh cabbages from each replication were chopped in quarters and freeze dried  at 80 °C and 5 mTorr after freezing with liquid nitrogen. Following freeze drying, the samples were ground and stored in a freezer at 20 °C until phytochemical analyses. Four biological replicates were used for the analyses of metabolites of ground cabbage powder.Winter cabbage  using the commercially employed method for producing cabbage juice. All harvested cabbages grown under open-field and bi-facial panels were pooled and juiced in a commercial juicer  to mimic commercial-level, bulk production. The heated cabbages were slowly cooled down for over 30 min and then squeezed. The filtered juice was packed in retort pouches and sterilized at 100 °C for 20 min. Pouches were stored at 4 °C in a refrigerator until use. Three analytical replicates were used for the analyses of metabolites of cabbage juice.Sinapis alba seeds) was added as an internal standard. The mixture was vortexed for 10 s and centrifuged at 1200× g for 10 min. The supernatant was decanted into a glass tube, and 1 mL of the supernatant was transferred to a 2 mL microcentrifuge tube. The lead barium acetate solution  was added to the tube, and the mixture was centrifuged at 15,000× g for 2 min. Following centrifugation, the mixture was drained into a poly-prep column filled with DEAE Sephadex A-25 . When the mixture was passed, 3 mL 0.02 M pyridine acetate, 3 mL deionized water, and 500 µL sulfatase solution  were added to the column sequentially, but not at the same time. The desulfo-glucosinolates were eluted using deionized water and filtered through a 0.22 µm syringe filter. The sample was injected into a high-performance liquid chromatography (HPLC)  equipped with a Kromail RP-C18 column  . The flow rate of mobile phase A (HPLC grade water) and mobile phase B (Acetonitrile) was 1.5 mL·min−1 with the following gradient system: 0 min, 1.5% B; 4 min, 4% B; 16.5 min, 25% B; 17 min 60% B; 19.1 min, 1.5% B; 24 min, 1.5% B. UV response factors were adapted to each glucosinolate for quantification [Glucosinolates in the cabbages were extracted and analyzed following a previously described method with little modification . Freeze-fication .g for 5 min. Following the centrifugation, 500 µL of the supernatant was transferred to a 1.5 mL PTFE tube. After which, 500 µL dichloromethane (DCM) with phenyl isothiocyanate (10 µg·mL−1) was added as a solvent for liquid–liquid extraction and internal standard, respectively. Then, the mixture was incubated at 37 °C for 4 h. After incubation, the mixture was shaken and centrifuged at 15,000× g for 3 min. The DCM layer was taken out into a vial with an insert, and 1 µL of the sample was injected into a gas chromatograph  coupled to a gas chromatograph–mass spectrometer  and an autosampler with an injector . The capillary column  was utilized for performing chromatographic separations. GC/MS conditions were as follows: injection and detection temperatures were set at 250 °C; flow rate of carrier gas was 1.5 mL·min−1; oven temperature was set starting at 35 °C reaching to 330 °C (increasing at the rate of 20 °C·mL−1); MS ion source and interface temperature were 300 °C and 250 °C, respectively; the mass scan range was 40–500 m/z.Freeze-dried cabbage powder (75 mg) was mixed with 1.5 mL deionized water. The sample was incubated for 10 min to extract hydrophilic glucosinolates and myrosinase. After extraction, the powder mixture and cabbage juice (1 mL) were centrifuged at 15,000× To assess the consumer acceptability of cabbage juice, we developed a testing methodology involving human subjects. This was reviewed by the Chonnam National University Institutional Review Board. Previously written book was referred for sensory evaluation . Seven d−1. The three mass spectrophotometry parameters, ion source, interface temperature, and mass scan range, were set to 300, 250 °C, and 40–600 m/z, respectively. For analysis of water-soluble metabolites, the initial oven temperature was set at 80 °C for 2 min, then increased by 12 °C·min−1 to reach 330 °C, and kept at 330 °C for 5 min at the end. For analysis of lipid-soluble metabolites, the initial oven temperature was set at 150 °C for 1 min, then increased by 12 °C·min−1 to reach 320 °C, and kept at 320 °C for 7 min at the end. Identification of metabolites was processed based on the database of the National Institute of Standards and Technology (NIST).Preparation of samples for profiling primary metabolites  followed previously described methods with some modifications ,35. To s−1. The flow rate of the carrier gas was 1.5 mL min−1 and both injector and detector temperatures were set to 250 °C. The three mass spectrophotometry parameters, ion source, interface temperature, and mass scan range, were set to 240, 310 °C, and 45 to 450 m/z, respectively. Amino acids were compared to authentic standards for identification. Norvaline was used as the internal standard.For quantification of free amino acids, an analysis kit for GC/MS  was used. Cabbage juice was added to 0.1 N HCl and amino acids were extracted at 25 °C for 10 min during shaking at 800 rpm. After then, amino acids in the supernatant were purified and derivatized according to the process delineated in the manufacturer’s manual. A capillary column  was used for separation. The oven temperature was kept at 110 °C for 1 min and increased to 320 °C at 26 °C minL*: lightness; L*: 0; L*: 97.13; a*: 0.05; b*: −0.76) boards. Cabbage juice was prepared in the same cup as the ones used for sensory evaluation, with a height of 5 cm from the surface. The sample from APV was used as a standard. The color difference (ΔE) was calculated using the following equation:Parameters for indicating color  were conducted using Prism 5 .The primary metabolites and amino acids data were normalized by appropriate internal standards for sample-wise normalization. The normalized data were used in principal component analysis (PCA) for obtaining a general outline of the variance of metabolites and information on differences in composition of metabolites among samples. Auto-scaling was applied to all variables for feature-wise (metabolites) normalization. Student’s −2·s−1 which was about 81% higher than the PPFD under APV. However, the differences between the systems in terms of mean air temperature and mean humidity were only 0.5 °C and 1.1%, respectively. Thus, one of the biggest differences in microclimate between APV and OF was sun radiation, which is consistent with the findings of a previous study [In the OF, mean air temperature, maximum air temperature, mean soil temperature, and cumulative growing degree days (CGDD) were 0.2 °C, 0.2 °C, 0.5 °C, and 19.9 °C, respectively, which were higher than those under APV . In addius study .p < 0.05) from those grown under APV (1450 ± 315 g), while yield reduction in mean weight was 156 g. Weight loss rate of cabbage grown under APV compared with OF was 9.7%. Cabbage diameter grown in OF (21.3 cm ± 3.1) was not significantly different (p < 0.05) from those grown under APV (21.1 cm ± 1.9). According to a review by Touil et al., [The formation of the cabbage head grown under OF cultivation was slightly denser than that grown under APV system A, while Brassica vegetables [p < 0.05). However, high-temperature-extracted cabbage juice showed a completely different composition and level of glucosinolates hydrolysis products compared to freeze-dried cabbage powder.It has been widely understood that glucosinolates and their hydrolysis products are strongly associated with consumers’ preference and health effects (including quinone reductase inducing activity and various anticancer activities) of getables ,20,36. Tgetables , which sgetables  even thogetables ,40,41. Ft-test, glucose, fructose, and all identified lipid-soluble metabolites  were significantly higher in OF than in APV, while malic acid, myo-inositol, sucrose, s-methyl-L-cysteine, and valine were significantly higher in APV than in OF. However, the difference in results between fresh cabbage and cabbage juice was caused by the difference between biological and technical replication, respectively. In case of commercial-level production, using pooled cabbage samples is a more realistic situation than processing cabbages from three biological replications separately. This is likely the reason for the difference in results between fresh cabbage and cabbage juice. In general, the method of biological replication has more fluctuation than technical replication due to deviation from various environmental factors. Therefore, analyzing products made by pooling cabbages from each treatment may be advantageous in comparing the quality differences of cabbage juice between the two systems.In total, 24 metabolites were identified using NIST library or authentic standards . To visup = 0.091). However, considering that half of panels received different sample configurations, there was no difference in set A  while there was a significant difference in set B . Among the subjects, 36 had experience in drinking cabbage juice, and those with experience distinguished the difference significantly (p = 0.034). These subjects, when asked how they were able to distinguish the odd sample out of the 2 OF and 1 APV samples, their answers were “strong taste”, “darker brown color”, and “overall strong feeling”. It was noteworthy that the subjects who received set B or have experienced cabbage juice before were able to discern; although, the difference was not significant with respect to the total number of people. Additionally, participants who distinguished set B gave higher scores in scent and flavor than set A . O’Mahony and Odbert., [The triangle test was adopted to check whether people can distinguish the difference between the two samples . In thisOdbert.,  showed tn = 36) scored 5.2, 4.7, and 5.1, with CI of 4.78–5.56, 4.29–5.05, and 4.60–5.56, respectively, suggesting color and taste were different. Subjects who had not experienced drinking cabbage juice (n = 34) scored 5.2, 4.3, and 4.5, and their CI was 4.79–5.64, 3.82–4.78, and 4.07–5.02, respectively, suggesting color and taste were different too. People who had experienced cabbage juice before tended to judge that the cabbage juice of OF is stronger than that of APV, than inexperienced people. However, regardless of the above results, preference between OF and APV sample was equal . This indicates that the difference observed by the subjects was not the critical level of difference for their preference.The result of the attribute test showed that the average scores for color, scent, and flavor were 5.2, 4.5, and 4.9, respectively, based on a 7-point hedonic scale . The CI p < 0.01, n = 3, technical replication). Pyrazines are found in roasted or toasted foods and are known to contribute to the roasted scent [Brassica vegetables owing to its property to increase while cooking and attribute unpleasant scents [Volatile organic compounds from glucosinolate hydrolysis analysis are shown in ed scent . Dimethyt scents . In varit scents . TherefoE* was 1.11 ± 0.06 (E* was from zero to one, and an experienced observer was able to differentiate from one to two [The value of Δ1 ± 0.06 B. The obe to two . In thisWe investigated possible differences in the quality of cabbage grown under OF and APV and its high-temperature-extracted juice, empirically. There were no differences of yield and metabolites between cabbage grown under OF and APV. However, cabbage juice made by pooled samples from each treatment showed differences in metabolites and consumers’ response to one set of triangle test. Given together, bulked commercial-level production of cabbage juice may show difference in metabolites; although, this difference was not significant enough to alter consumers’ preference on the cabbage juice . To the


Plantago lanceolata. The ACE and Ag. NPs were characterized and assessed their biological and antioxidant activities. The existence of nanoparticles (NPs) was confirmed by color shift, atomic force microscopy (AFM), and UV–Vis’s spectroscopy. The FT-IR analysis indicated the association of biomolecules (phenolic acid and flavonoids) to reduce silver (Ag+) ions. The SEM study demonstrated a sphere-shaped and mean size in the range of 30 ± 4 nm. The EDX spectrum revealed that the Ag. NPs were composed of 54.87% Ag with 20 nm size as identified by SEM and TEM. AFM has ended up being exceptionally useful in deciding morphological elements and the distance across of Ag. NPs in the scope of 23–30 nm. The TEM image showed aggregations of NPs and physical interaction. Ag. NPs formation also confirmed by XPS, DRS and BET studies. Ag. NPs showed efficient activity as compared to ACE, and finally, the bacterial growth was impaired by biogenic NPs. The lethal dose (LD50) of Ag. NPs against Agrobacterium tumefaciens, Proteus vulgaris, Staphylococcus aureus, and Escherichia coli were 45.66%, 139.71%, 332.87%, and 45.54%, with IC50 (08.02 ± 0.68), (55.78 ± 1.01), (12.34 ± 1.35) and (11.68 ± 1.42) respectively, suppressing the growth as compared to ACE. The antioxidant capacity, i.e., 2,2-diphenyl-1-picrylhydrazyl (DPPH) of Ag. NPs were assayed. ACE and Ag. NPs achieved a peak antioxidant capacity of 62.43 ± 2.4 and 16.85 ± 0.4 μg mL−1, compared to standard (69.60 ± 1.1 at 100 μg mL−1) with IC50 (369.5 ± 13.42 and 159.5 ± 10.52 respectively). Finally, the Ag. NPs synthesized by P. lanceolata extract have an excellent source of bioactive natural products (NP). Outstanding antioxidant, antibacterial activities have been shown by NPs and can be used in various biological techniques in future research.Silver nanoparticles (Ag. NPs) have shown a biological activity range, synthesized under different environment-friendly approaches. Ag. NPs were synthesized using aqueous crude extract (ACE) isolated from  The applications are primarily determined via properties such as size, shape, composition, crystallinity, and structure of the NPs3. The synthesis and characterization of NPs is an emerging nanotechnology field established over the past two decades due to a wide variety of applications in the areas of chemistry, physics, biology, medicine, textile5, wastewater treatment, and drug delivery6. Generally, the synthesis of silver nanoparticles (Ag. NPs) can be modulated by an extensive range of aspects, such as biological substances, temperature, pH, the strength of metal ions, heat, reaction time, and electromagnetic waves7.Nanoparticles (NP) are synthesized from fauna/flora extracts and elements such as silver (Ag), gold (Au), platinum (Pt), nickel (Ni), zinc (Zn), copper (Cu), and compounds like titanium dioxide , received growing attention among metal NPs9. Due to the exceptional features such as their antibacterial, antiviral, antifungal, and anti-inflammatory activities, Ag. NPs gained unlimited attention from scientists11. It may be used in nanocomposite products, cosmetics, food processing, computer modules, etc. It also has high chemical consistency, excellent biocompatibility, catalytic efficiency, low toxicity, and eco-friendliness compared to other outmoded antibiotics and chemicals. Ag. NPs are also used in diagnosing various acute and chronic diseases, pharmaceutical delivery, electronics, bio-sensing, the food industry, the garment industry, paints, and health supplements12.The adaptations and modifications of these factors can therefore regulate and modulate the characteristics of Ag. NPs, including scale, shape, surface load, surface properties, and crystalline nature, as well as bioactivities13. Various techniques such as Ag ions reduction in an aqueous medium with or without stabilizing agents can synthesize NPs14, in organic solvents, by thermal decomposition15, chemical reduction by radiation16. In the literature, chemical reduction and photochemical reduction of quashing micelles have also been reported18. Typically, these approaches are expensive and require hazardous chemicals that can cause biological risks. Biological techniques of preparing Ag. NPs using microorganisms21, fungi22, enzymes23, and plant extracts have been recommended to substitute chemical and physical methods due to their eco-friendly nature. Synthesis of NPs by using plants or plant extracts is more accessible than other approaches to a significant degree by reducing the elaborate processes of microbial culture maintenance26.Compared to the same material in the bulk state, nanomaterial sizes ranging from 1 to 100 nm have significant differences in their properties27. Ag was the third metal the ancient civilizations learned to use after Au and Cu about 4000 BC. Before the invention of antibiotics, Ag was the most significant antimicrobial substance accessible and various forms for preserving foodstuffs28. Ag utensils in the kitchen have been prevalent worldwide due to knowledge of antibacterial activities29. Plants and microorganisms are now used for NPs synthesis. Natural bioactive compounds, i.e., primary and secondary metabolites, have a wide range of applications. Chemists have been investigating strategies for eliminating harmful compounds and utilizing healthy metabolites30.The antimicrobial potential of Ag has been recognized long beforePlantago lanceolata is a perennial medicinal plant belonging to the family Plantaginaceae with about 265 species31. Other plants such as Artocarpus heterophyllus13, Sesbania grandiflora30, Prosopis juliflora, and Semecarpus anacardium extract are used for the synthesis of NPs with different metals32. Due to its abundance of active secondary metabolites, P. lanceolata is used as an herbal plant in many countries. This plant addresses several health issues, including skin problems, like abscesses and pimples33. It is also used to treat cancer, embolism, bee bites, Tuberculosis (TB), and other pneumonia-related anomalies. The plant species and other members displayed promising antimicrobial activity against bacteria, fungi, anthelmintic, and other microbes. It has gained a lot of popularity in countries like Turkey and the Indian subcontinent because of its antioxidant and antiviral activities. Infectious diseases like diarrhea, dysentery, and colic malaria can be treated with P. lanceolata boiled water extract with honey35.P. lanceolata. Mainly tannins, flavonoids, mucilage, silica, zinc, potassium salts, iridoid glycosides, phenylpropanoid glycoside, acteoside, flavonoids, and phenyl carboxylic acid are found in leaves. Catapol and its precursor aucubin are the most common iridoid glycosides in P. lanceolata36. Plantago lanceolata is widely used to treat blisters, stomach disorders, uterine cancer, pimples, homeostatic, TB, asthma, bronchitis, and diarrhea40. The secondary metabolites present in the leaves, roots, and bark are responsible for therapeutic potential. Plantago lanceolata is local to Europe and Asia, but with a suitable environment, it spread to North America and many other areas41.There are several other reactive chemical metabolites in Plantago lanceolata and its other family affiliate are the best source of several active and beneficial metabolites source and used against several diseases from ancient times. Here we synthesized its Ag. NPs with the ACE enhance its potential against the various enzymes and microbes causing diseases. Ag. NPs have an enormous surface area for their activities against their target compared to simple extracts, which play an essential role in Ag. NP synthesis40. The aqueous crude extract (ACE) contains many active compounds, such as tannins, terpenes, anthraquinone, sugar, glycosides, phlobatannins, emodin, phenolic acid, flavonoids, and saponins42. The reduction of Ag is actively involved in these compounds for NPs synthesis. The current study aims to prepare NPs of low-cost to establish an innovative approach using different fractions of methanolic crude extracts of P. lanceolata and compare their antibacterial and antioxidant activity with ACE.Plantago lanceolata was collected from the Mansehra , Khyber Pakhtunkhwa, Pakistan  was filtered, and a rotary evaporator was used to get a thick slurry, stored in the refrigerator at 4 °C. The CME was subdivided on solvent polarity base into n-Hexane, dichloromethane, ethyl acetate, and ACE.The tan Fig. . Before n-Hexane, Sodium borohydride, DPPH, and other micro-apparatus were purchased from Davidson and Hardy Ltd, Ireland. The King Fahad Medical Research Centre, King Abdul Aziz University, Jeddah, Saudi Arabia, provided bacterial isolates. Deionized water was used in the process for solution preparation, as well as washing and rinsing laboratory equipment. To reduce humidity, the equipment was wrapped in aluminum foil during the tests.Methanol, Ethanol, Ethyl acetate, P. lanceolata’s extract mixed with 1 Mm AgCl solution (1:9 ratio) at room temperature. Via integrating the freshly made P. lanceolata extracts with the freshly made 1 mM AgCl2 solution (1:9 ratio) at room temp, Ag. NPs were synthesized in ice-cold 100 mL deionized water. The ACE stock solution was prepared by mixing 0.1 g of extract into 50 mL of methanol and filtering with Whatman Grade 42. The two stock solutions were combined in a 250 mL flask in various combinations and stirred at room temperature for 4 h. Two solutions were mixed in a 1:1 proportion for the successful synthesis of Ag. NPs6. Through incubation, changes in color from pale yellow to reddish-brown were noted in the formation of the NPs. The mixture was then centrifuged at 15,000 rpm for 1 h after incubation to isolate the NPs, and the NPs obtained were air-dried and then further cleaned with deionized water and stored in the incubator. The collected powder (NPs) was stored in the refrigerator for further characterizations7.Wet chemistry procedures have been used to make nanoparticles, which entailly creating the particles in a solution, drop projecting them onto a substrate, and then extracting the solvent, surfactants, and other components from the particles. Freshly prepared 44. The size and morphology of Ag. NPs were performed by atomic force microscopy (AFM) (AGILENT-N9410A series 5500).The NPs obtained were characterized by various measurements, such as UV–Vis spectroscopy, to calculate the maximum wavelength for the quantitative determination and optical properties. The UV–Vis spectrum of biosynthesized Ag. NPs and ACE were determined using the UV–Vis 1800 spectrophotometer . The instrument was operated at room temperature with a 1 nm resolution in the 200, and 600 nm scalesλ range of 400 to 4000 cm−1 and 4 cm−1. The universal disc method was followed for the analysis6.The PerkinElmer FT-IR 1600 spectrophotometer (USA) was used to compare the IR spectra of both Ag. NPs and ACE in the α1 + K α2 radiation (λ = 0.15425 A), the phase units of Ag. XRD measured NPs. The Bragg angles vary from 1° to 60° at a scanning rate of 2° min−1 using (Eq. Using Rigaku Smart Lab II X-Ray powder diffraction (Japan) with an average source of Cu K λ is the wavelength of X-rays and β is the line broadening at half the maximum intensity (FWHM) in radians13.I was using the ‘Scherrer equation’ Eq. , from th−1 and sonicated using a sonicate bath. The sonicated stock solution (1 mg mL−1) was diluted 20 times to measure the size of Ag. NPs. Then the sonicated aqueous solution was dried by taking one drop of it on a glass plate. Finally, the sample was placed on a carbon-coated copper grid, and images were taken45. The Oxford-EDS system (UK) was used to conduct energy dispersive X-rays (EDX) spectrometry on the ACE and Ag. NPs.Morphology of Ag. NPs were examined by using SEM ZEISS SEM (Germany) with a magnification (Max); 300,000X and resolving Power (Max); 2.3 nm that contains information about the surface topography and composition of the sample. The Ag. NPs were suspended in deionized water with a concentration of 1 mg mL20.The microstructure and particle size of the Ag. NPs were studied using TEM . The sample was dissolved in ethanol and sonicated to disperse. The pieces were then coated in carbon films, and images were takenFor the diffuse reflection spectroscopy (DRS) of the Ag. NPs, the absorption spectra of biosynthesized NPs can be calculated by using Tauc’s relation Eq.  to deterα” signifies the absorption coefficient and “B” is a constant. The terms hυ represents the photon energy and Ecb signifies the band energy. The values of band gap energy can be determined from the linear part of the curve (between (hυ)2 versus hυ).In this equation “The surface area and pore volume of Ag. NPs were measured at 77 K using the Brunauer–Emmett–Teller (BET) method.X-ray photoelectron spectroscopy (XPS) with a monochromatic Al Kα excitation source (VG Scientific ESCALAB220i-XL). Following Shirley background subtraction, CasaXPS software was used to do quantitative analysis of XPS data. Using a PHI5000 VersaProbeIII (Japan) analyser, all XPS data were compared to a standard binding energy of C1s of 284.5 eV , Proteus vulgaris (gram-negative), Agrobacterium tumefaciens (gram-negative), and Escherichia coli (gram-negative), were used for the antibacterial activity Ag. NPs and ACE of P. lanceolata. These bacterial species were cultured for 24 h in the Petri dishes in the nutrient broth medium using up to 20 mL of melted and cooled nutrient agar7. The four tested species were simmered over the nutrient agar media, the ACE, and Ag. NPs were placed in the well over the medium, already developed using sterile borer. 100 μL of each 10, 50, and 100 μg mL−1 concentration of Ag. NPs were loaded for each well. Furthermore, the effects of ACE and Ag. NPs were studied as a function of time in fixed volumes of bacterial strain cell morphology, and a parallel bacterial inhibition was observed on the solid agar nutrient substrate. The percent inhibition zone was recorded for each concentration, and the LD50 and IC50 value was determined from the percent inhibition. As a reference, ampicillin was used46.Bacterial strains, namely 50 was also calculated. Then layer chromatography was done for the Ag. NPs and observed before and after DPPH spray. A stable free radical at 517 nm using DPPH was used to evaluate the radical scavenging activities of the ACE and Ag. NPs. It is reduced due to H or electron donation by 1,1-diphenyl-2-picrylhydrazyl, whose color changes from violet to orange-yellow. The experiments were conducted in triplicate. Briefly, 1 mL solution of DPPH (1 mM) prepared in methanol was mixed with 3 mL samples . The solutions were stored for 30 min in the dark and the absorption was estimated at 517 nm47.For antioxidant activity, the DPPH radical scavenging analysis, hydrogen atom or electron donation abilities of the corresponding extract , and ascorbic acid (standard drug) was carried out through antioxidant action and ICPlantago lanceolata) were carried out in accordance with relevant institutional, national or international guidelines or regulation.Experimental research and field studies on plants (either cultivated or wild), comply with relevant institutional, national, and international guidelines and legislation. All plant studies , after taking permission from Forestry, Environmental and Wildlife, Khyber Pakhtunkhwa, Pakistan.The Before collection, the plant was identified by Dr. Hanif Khan (Taxonomist), using the standard protocol at the Department of Life Science, Northwest University, Xi’an, China.We all declare that manuscripts reporting studies do not involve any human participants, human data, or human tissue. So, it is not applicable.6. Efforts were made to test P. lanceolata for NPs synthesis and its activity3. The development of Ag. NPs of comparable particle size were observed by the surface plasmon resonance (SPR). Color shift was visual proof for converting Ag+ ions of AgCl2 into Ag. NPs with A.C.E. Ag. NPs had dark yellowish-brown color in an aqueous solution due to the surface plasmon resonance phenomena. After combining aqueous fraction and AgCl2 colorless solutions in a 1:1 ratio, the color change due to Ag’s oxidation in NPs48. The color strength improved over time, and after 4 h of constant stirring, the color of the mixture turned reddish-brown52.Green biosynthesis of nanoscale materials by biological materials was promoted to reduce toxic component production53. At wavelength 432 nm, the UV–Vis spectrum gave maximum absorbance , indicating the contribution of these groups  in the synthesis of Ag. NPs45.The Ag. NPs, FT-IR spectrum is presented in Fig. The design, morphology and size of the orchestrated silver nanoparticles were described by the AFM pictures. Figure 5. Entire reflection peaks corresponded to Ag. NPs with face-centered cubic symmetry53. The high intensity of peaks showed that the Ag. NPs were highly crystalline15. The diffraction pattern of Ag. NPs indicated peaks at 23.52°, 27.83°, 38.23°, 44.25°, 46.23°, 54.80°and 57.50° of face-centered cubic pure Ag. NPs (JCPDS 76-1393)55. It verified that the Ag. NPs’ principal constituent was Ag metal. The calculated size of Ag. NPs was 26.8 nm at 2θ = 27.70°, 26.5 nm at 2θ = 32.10° and 28 nm at 2θ = 46.10°55. The particle size obtained from the XRD plot using  of the element in Ag. NPs Fig. f59. The α” signifies the absorption coefficient and “B” is a constant. The terms hυ represents the photon energy and Eg signifies the band energy. The values of band gap energy can be determined from the linear part of the curve (between (hυ)2 versus hυ). The calculated amount of band gap energy was 2.34 eV for Ag. NPs as shown in  gas is used in the BET surface area analysis of Ag. NPs. The reason is that N2 gas is available in pure form and it has very interaction with solid Ag. NPs. The BET adsorption isotherms for Ag. NPs is described in . The result is associated with the area of the inhibition assay. The Ag. NPs showed very significant activity against all the four selected bacterial strains  as compared to ACE, showing the highest resistance of P. vulgaris among all the four strains30. Also show good results for IC50 values for (P. vulgaris (55.78 ± 1.01), A. tumefaciens (08.02 ± 0.68), E. coli (11.68 ± 1.42) and S. aureus (12.34 ± 1.35)  the overproduction of reactive oxygen species like superoxide (O2−) and OH−, (ii) the existence of Ag+ ions in Ag. NPs that form bacterial protein sulfhydryl groups, and (iii) the release of Ag+ ions by Ag. NPs that infiltrate the cellular structure and destroy bacteria62.The antimicrobial activity of Ag. NPs were enhanced by the phytonutrients that preserved them and the increased antibacterial potency of Ag. NPs is due to the macromolecules and surface area. The small size, dispersion, and spherical morphology may be correlated with the antibacterial viability of the Ag. NPs, which offer a wide surface for maximal contact with bacteria, resulting in even more damage than larger particle sizeies Fig. 63. Effec 55.78 ± .01, A. t45. The redox study indicates that Ag+ ions emitted from the Ag. NPs surface is responsible for their antibacterial assay19. The thin peptidoglycan framework of bacteria ensures that the Ag. NPs can directly enter the cytoplasm and interact with the cell membrane, causing damage to membranous protein57. Also, Ag. NPs posed significant antibacterial efficacy against Escherichia coli. Escherichia coli are much more susceptible to Ag. NPs than Agrobacterium tumefaciens, Proteus vulgaris, and Staphylococcus aureus. The smaller NPs have a reduced interparticle gap and an increased number of unit cells measured from the XRD measurements, impacting the electron density at the surface of the NPs64. Thus, Ag. NPs demonstrated the most vital antimicrobial property based on our findings which indicate that the choice of P. lanceolate is best for the synthesis of Ag. NPs.Ag. NPs have a large surface area and smaller size, allowing broad interaction with the bacteria cell wall and toxic to cytoplasmic content in a systematic way65. Both samples were applied at different concentrations (10–100 μg mL−1). The thin layer chromatography of DPPH identified zones with varying values of retention factor (rf)66. ACE and Ag. NPs have substantial DPPH free radical scavenging activities to estimate their antioxidant ability, which is proven relative to the standard antioxidants44. The ACE and Ag. NPs exhibited maximum antioxidant capacity of 62.43 ± 2.4 and 16.85 ± 0.4 at 100 μg mL−1, respectively) compared to control (69.60 ± 1.1 at 100 μg mL−1)67. Also gave results for IC50 values for ACE and Ag. NPs 369.5 ± 13.42 and 159.5 ± 10.52 respectively regarding DPPH radicals . High antibacterial activity for Ag. NPs against