Beta vulgaris ssp. vulgaris), our understanding of the genomic diversity and functional genes in its cultivars is limited, resulting in slow breeding progress. To address this issue, a total of 306 germplasm materials of major cultivars and breeding lines from China, the USA, and Europe were selected for genome resequencing. We investigated population structure and genetic diversity and performed selective scanning of genomic regions, identifying six novel genes associated with important agronomic traits: the candidate genes DFAX2 and P5CS for skin roughness; the candidate genes FRO5, GL24, and PPR91 for root yield and sugar yield, and the pleiotropic candidate gene POLX for flourishing growth vigour, plant height, crown size, flesh coarseness, and sugar yield. In addition, we constructed a protein–protein interaction network map and a phenotype-gene network map, which provide valuable information for identifying and characterizing functional genes affecting agronomic traits in sugar beet. Overall, our study sheds light on the future improvement of sugar beet agronomic traits at the molecular level.Due to the relatively brief domestication history of sugar beet ( Beta vulgaris spp. vulgaris L.) is a core eudicot of the order Caryophyllales, family Amaranthaceae (formerly Chenopodiaceae), and subfamily Betoideae2. With an estimated genome size of 714–758 Mb, conventional sugar beet has 2n = 18 chromosomes3. It shares a common paleohexaploid ancestor with Rosaceae and Asteraceae4. Sugar beet is a major biennial root crop in temperate regions and is the world's second largest sugar crop after sugarcane. Global cane sugar production in 2021 was 130.964 mln tons, with sugar beet cultivation of 4,628,697 hectares producing 34.204 mln tons of sugar, or approximately 26% of global sugar production . In addition, sugar beet is a source of bioethanol and animal feed5. Due to the abundance of wild resources and the continuous development of new varieties, the yield, quality and disease resistance of European and American sugar beet varieties have steadily improved. However, compared with foreign varieties, Chinese varieties have lower genetic diversity and limited breeding resources. Therefore, making good use of the existing core germplasm resources for pre-breeding and variety improvement is a priority for improving sugar beet yield, quality and disease resistance.Sugar beet  genome sequence has been published18, and the assembled genomes of wild-derived beet accessions have become available20. However, our knowledge of the sugar beet genome and its functional genes associated with important agronomic traits is still limited compared to, for example, that for corn, soybean and rice. This is due to the complex genetic background and long reproductive period of sugar beet. Therefore, many sugar beet varieties need to be further investigated at the genomic level, especially for molecular traits associated with potential genetic networks for important agronomic traits such as root yield and sugar content.Genome-wide association study (GWAS) is a genome-wide polymorphism assay for genetic variation that uses a natural population or a collection of different individuals to obtain genotypes, which are then statistically analysed against a phenotype to determine whether a genomic variant is associated with a trait of interest. GWAS has become a standard approach for deciphering genotype–phenotype associations in many species due to the development of rapid genotyping and next-generation sequencing technology21. This information could serve as a valuable reference for molecular breeding and evolutionary studies in sugar beet, potentially supporting the development of new sugar beet varieties22.In this study, the genetic diversity of core germplasm resources from four growing regions representing three major ecotopes in northern China was examined, and association analysis was conducted on important agronomic traits related to yield and quality. We resequenced 306 sugar beet accessions from four growing regions representing three major ecological zones in northern China and performed genotypic and phenotypic analyses. Our major objective was to explore the dynamics of genomic variation in response to population selection in sugar beet from the three major ecological regions of northern China. In parallel, we conducted a GWAS of 26 phenotypic traits in this species. The aim of this study was to identify and characterize multiple patterns and genes associated with essential agronomic traits. Our results provide important insights into the genome structure of sugar beet, including novel genes associated with important agronomic traits and genetic networkshttp://plants.ensembl.org/Beta_vulgaris/Info/Index). The resulting high-density and high-quality genotype data covered 566,181,630 bp with an average sequencing depth of 11 × and a maximum depth of 22.5 × per accession. The average alignment rate for the population samples was 96.43% ± 0.5%, with an effective localization rate of 74.8%. The average sequencing depth across the genome (excluding gap regions) was 12.2 ×  ± 1.1 × . The genome coverage was 86% ± 1.8%. The high mapping and coverage rates ensured the reliability and high quality of the sequencing data. This comprehensive dataset allowed for a more accurate assessment of genetic diversity and identification of potential candidate genes associated with agronomic traits in sugar beet accessions.In this study, we performed high-depth genome-wide resequencing of 306 sugar beet accessions using an Illumina HiSeq 2000 sequencer, obtaining 1977.12 Gb of sequencing data. This collection included 72 endemic accessions from Northeast China (Harbin), 114 endemic accessions from North China (Hohhot), 100 endemic accessions from Northwest China (50 from Urumqi and 50 from Shihezi), and 20 accessions from abroad. This diverse population represents the genetic diversity of ecotypes, geographical locations, and various traits. After filtering the raw sequencing data , the high-quality clean data were aligned to the sugar beet reference genome, RefBeet-1.2.2  loci and 3,975,247 insertion/deletion (INDEL) loci. After screening for a gene frequency  greater than 0.05, 8,258,753 SNPs were identified. Among them, 872,623 were located 1 kb upstream of the genes, and 512,777 variants were in exonic regions, resulting in 242,816 non-synonymous mutations and 269,961 synonymous mutations. The ratio of non-synonymous SNPs to synonymous SNPs was 0.899. A total of 2,961,245 variants were found in intronic regions, 3309 variants were in splice sites, and 748,749 variants were upstream of one gene and downstream of another gene. Additionally, 12,439,140 variants were located in intergenic regions  method, and the diversity of genotypes among these materials was assessed. The 306 accessions were divided into four groups based on the phylogenetic tree and genetic distance Fig. A. The fiWe conducted a structure analysis on 306 sugar beet samples using Admixture software to gain insights into the genetic makeup and relationships of the germplasm resources. Each column in the image represents one individual, and the length of the differently coloured segments indicates the proportion of an ancestor in the individual's genome Fig. C. When KPrincipal component analysis (PCA) showed that some samples from group 2 were more clearly separated from the other three subgroups in the upper left corner, group 1 and group 3 both radiated to the lower left, group 4 samples were more centrally distributed, and only a few samples deviated from the other three subgroups and diverged to the lower right. With respect to the different production areas, samples from the Harbin region were tightly clustered. In contrast, samples from the Hohhot region showed the greatest dispersion, and the few samples from the Urumqi production area showed significant separation  analysis showed that the LD decay distances of the four subpopulations were not significantly different, with G2 > G1 > G3 and G4  analysis determines the concordance of all genetic markers to reflect the degree of correlation among individuals. This analysis revealed that group 2 was the smallest among the four subgroups, while groups 1, 3, and 4 were convergent  was calculated to represent the genetic distance and differentiation between populations. The Fst values between the four subpopulations and the four production areas showed that G3 and G4 had the lowest values, with little differentiation and a close genetic distance. The Fst values of G2 and G4 were the highest, indicating the greatest degree of differentiation between these two subgroups  in three major ecological regions . Differences between traits were determined by calculating Pearson correlation coefficients of the traits  greater than 0.05. The GWAS of the 26 agronomic traits was performed using Genomic Association and Prediction Integrated Tool (GAPIT) with the maximum likelihood method mixed linear model (MLM), and Manhattan and QQ plots were generated  and physiological components . Due to this characteristic of sugar beet, the present study introduces the complex trait of sugar yield as a means to investigate yield quality. For the sugar yield trait, a total of 10 candidate genes were identified. After further analysis, three genes, FRO5 (BVRB_3g053570), GL24 (BVRB_3g053550), and PPR91 (BVRB_1g005550), two on chromosome 3 and one on chromosome 1, were found to be associated with both root yield and sugar yield The five traits associated with sugar yield  shared a common gene, POLX (BVRB_1g140620), located on chromosome 6 Fig. . PhenotyThe results reveal that the functional mutations formed three haplotypes, AA, GA, and GG, for each gene. Comparisons of haplotypes and phenotypes separately showed that for flourishing growth vigour and plant height, the phenotypes of GA and GG displayed a highly significant correlation; for crown size and flesh coarseness, AA, GA, and GG all exhibited a highly significant correlation; and for sugar yield, the phenotypes of GA and GG exhibited a significant correlation Fig. B. FollowThrough an in-depth exploration of the GWAS results, we identified instances where one trait was associated with multiple genes, and one gene was linked to multiple traits. Furthermore, we discovered complex networks between various phenotypes and genes due to extensive protein-level interactions among genes. To gain further insights, we performed a functional mutation-based haplotype test to construct a protein–protein interaction (PPI) network map and a phenotype-gene network map, which included 14 traits and 256 annotated pleiotropic genes Figs. , 8. We cIn this genetic network, the six trait categories were linked by 14 central nodes containing 256 genes, with the largest central node being a root trait. In the PPI network diagram, the genetic network was dominated by ten major genes that linked different pleiotropic genes. The gene PPI network diagram allowed the identification of the pleiotropic genes of the major nodes. The combination of phenotypic associations and protein interactions with these primary genes provided a basis for identifying important candidate genes. This comprehensive information could be utilized to develop targeted breeding strategies, enhance the efficiency of genetic selection, and accelerate the process of cultivar development.In this study, we conducted the first resequencing of 306 sugar beet germplasm accessions, providing valuable insights and supplementary data for sugar beet genome research. The selected materials in this study encompass resource materials from four major production areas in China. We performed the first-ever analysis of the genetic structure of germplasm materials from these critical production areas using population genetics and GWAS, determining the genetic diversity levels and differentiation degree among them. Our results demonstrated that despite different regions and breeders selecting the same materials through years of crossbreeding, these materials exhibited varying degrees of domestication while still maintaining similar genetic backgrounds.We assessed the population classification of the resource material and examined their relationships with the domestication levels in other regions. Our comprehensive analysis incorporated a phylogenetic tree, PCA, and LD analysis. Our results showed that, in contrast to those from the other three production areas, the materials from the Hohhot region exhibited a high degree of domestication, a high level of differentiation, and reduced genetic diversity. We observed a trend where varieties with similar domestication levels tended to exhibit greater genetic similarity. However, varieties with similar genetic backgrounds still displayed differences in geographic distribution and domestication levels, implying that gene exchange might have occurred between germplasm of different populations. This phenomenon reflects the domestication history of sugar beet resource material in the four major planting regions after their introduction to China and highlights the increasing complexity of the sugar beet genetic background due to years of hybrid selection. Population structure analysis revealed a simpler genetic background in the northeastern region than in the other regions, suggesting that the resource materials derived from this area had a more homogeneous ancestral origin.IBS, θπ, and Fst analyses revealed relatively consistent genetic diversity levels among individuals within different subpopulations and production areas. However, a higher degree of correlation was observed among individuals from the North China production area, suggesting that the germplasm resource materials from this region experienced multiple recombination events among individuals. Fst analysis indicated that the resource materials from Northeast and North China were the most differentiated. The genetic proximity and the lowest degree of differentiation suggest that the resource materials from the Northeast and Northwest production areas, particularly Harbin and Urumqi, have experienced more frequent genetic exchanges with each other.This research revealed pleiotropic genes associated with skin roughness and sugar yield, which can be used to screen new candidate genes for future sugar beet breeding programmes. This information is very helpful for breeders to develop improved sugar beet varieties with desired traits, ultimately enhancing crop performance and overall production efficiency  absorption in roots. Cu is an essential trace element required for various biological processes, and it has been selected as a cofactor of various protein active centres over the course of evolution due to its unique chemical properties37. Further biochemical analysis and confocal imaging of Cu-specific fluorophores showed that FRO5 and FRO4 were involved in the absorption of high-affinity root Cu, specifically reducing Cu(I) by Cu(II), which caused a defect in iron transport from root to stem related to the lack of iron oxidase activity. These findings provide insight into the roles of FRO5 and FRO4 in regulating Cu and iron uptake in plants and suggest their potential use as targets for improving crop growth and yield.A study of Arabidopsis spl7 mutants revealed that FRO5 and FRO4 transcription levels increased significantly in an spl7-dependent manner when Cu was deficient38.The second gene was GL24, which encodes germin-like protein subfamily-2 member-4. Germin-like proteins (GLPs) are ubiquitous plant proteins that have been implicated in various biological processes, including the cold resistance response in rice39. PPR-related mutants have been observed in several plant species, such as Chlamydomonas40. Arabidopsis41, maize42, turnip43, and Sorghum44. Further investigation is needed to determine the exact role of PPR91 in sugar beet and its potential application in breeding for improved sugar production.Finally, we identified the PPR91 candidate gene in our study, a member of the pentatricopeptide repeat (PPR) protein family, one of the largest and most complex protein families in plants. PPR proteins have been found to be involved in various crucial functions related to organelle biogenesis and plant development45. The PPR protein family plays an essential role in regulating gene expression related to cytoplasmic male sterility (CMS)46, participates in the regulation of embryo formation47, and influences plant growth and development48. In our study, we found a strong correlation between the candidate gene PPR91 and sugar production. Further investigation is needed to explore the precise mechanism by which this gene regulates sugar production in sugar beet plants.The PPR protein family is characterized by a degenerate 35-amino acid repeat sequence, with each peptide arranged in a series with 2–27 repeats. Many PPR proteins are predicted to target either mitochondria or chloroplasts. reported that disease resistance protein (TIR-NBS-LRR class), retrovirus-related Pol polyprotein from transposon TNT 1–94, and transposon Ty3-1 Gag-Pol polyprotein were targets of miRNA482/2118 members, and they could affect the expression of nucleotide binding site leucine-rich repeat (NLR) mRNAs and disease resistance49. The involvement of POLX in multiple traits suggests a potential role in regulating disease resistance and yield in sugar beet. In summary, the annotated functions of the genes related to sugar production are associated with stress resistance and disease resistance.We also identified a gene, POLX, among the genes related to sugar yield that was associated with multiple traits. POLX is annotated as a retrovirus-related Pol polyprotein from transposon TNT 1–94. Canto-Pastor et al50. Plants have developed sensing and response mechanisms to adapt to adverse environmental stresses such as drought, high salinity, and low temperature. As an osmotic protector and molecular switch, sugar regulates the resistance and adaptability of plants to stress51. It is hypothesized that the identified genes not only participate in the regulation of disease resistance and stress resistance but also play a role in regulating sugar production.To grow and reproduce, many pathogens have evolved mechanisms to obtain glucose by hijacking the host's sugar outflow system. As a result, pathogenic bacteria can alter the sugar efflux of infected sites and modulate plant immunity52. However, selecting for favourable traits could inadvertently lead to the selection of undesirable traits since multiple traits can be interrelated. Understanding the genetic networks underlying different traits could benefit breeders in terms of enhancing breeding efficiency by helping them identify similarities among various genetic resources. For example, in recent decades, functional genomics in rice has rapidly developed, and some key genes controlling yield and grain quality have been identified53. Such efforts have enabled breeders to select and improve multiple related traits simultaneously, which is the ultimate goal of plant breeding.The primary objective of plant breeding is to combine several desirable traits into a single genome, which requires breeders to enhance and select multiple interdependent traits simultaneouslyIn this study, we established a novel phenotypic-gene network that includes 14 traits and 247 annotated genes. The establishment of a phenotypic-gene network, as achieved here, can provide a more direct understanding of the relationships between genes and phenotypes, facilitating the discovery of important candidate genes for improving yield quality traits in sugar beet at the molecular level. By identifying these key genes, breeders can more precisely select desired traits while avoiding the selection of unwanted traits, ultimately leading to the development of more productive and sustainable crops.http://www.cgris.net). Phenotypic correlation analysis using the R environment (gWQS package) was performed on the collected phenotypic data. Seeds were planted in pots and grown in a climate chamber  at 25 ± 1 °C in day/20 ± 1 °C in night (14 h day/10 h night) with 70% relative humidity until the 6-true leaf stage (approximately 4 weeks old). Seedling leaves were used for qRT-PCR validation.The experimental materials for this study were provided by the College of Modern Agriculture and Ecology of Heilongjiang University, the Institute of Specialty Crops of the Inner Mongolia Autonomous Region Academy of Agricultural and Animal Husbandry Sciences, the Sugar Beet Research Institute of the Shihezi Academy of Agricultural Sciences, and the Economic Crop Industry Research Center of the Xinjiang Provincial Academy of Agricultural Sciences. The 306 materials were planted in four different locations with different environmental conditions in Northeast, North and Northwest China for phenotypic data collection. Seventy-two materials were planted at the experimental site in Hulan District, Harbin, Heilongjiang Province, Northeast China; 134 materials were planted at the experimental site in Hohhot , Inner Mongolia Autonomous Region, North China; 50 materials were planted at the experimental site in Urumqi, Xinjiang Province, Northwest Region and 50 materials were planted at the experimental site in Shihezi  according to the manufacturer's protocol. For each landrace, a single individual was used for genome sequencing on the Illumina HiSeq PE150 platform. Library construction and sample indexing were performed as described.54. Finally, data from three biological replicates are presented as mean ± standard deviation (SD). The result was shown in Supplementary Table Total RNA was isolated from each sample using polysaccharide polyphenol plant TRIzol Universal Extraction Reagent  according to the manufacturer’s protocols. The purified RNA was stored at − 80 °C until further analysis. First-strand cDNA synthesis was performed using 5X Evo M-MLV RT Reaction Mix reverse transcriptase according to the manufacturer’s protocols . qRT-PCR was performed using 2 × SG Green qPCR Mix  and a real-time PCR machine (ABI No. QuantStudio 7 Flex USA) according to the manufacturer’s protocols. The primer sequences used in the qRT-PCR were shown in Supplementary Table 55 (version: 0.7.8). This was carried out using the command line "BWA mem -t 4 -k 32 -M". After alignment, the sorted reads were run through the 'rmdup' program to remove any PCR duplicates. At this stage, only read pairs with the best mapping quality and identical external coordinates were retained.The sugar beet reference genome (RefBeet-1.2.2) was used to align paired-end reads with Burrows-Wheeler Aligner56. Allele frequencies in the sample were calculated using a Bayesian method, and genotype likelihoods were determined based on the readings for each individual at each genomic site. We used the parameters '-q 1 -C 50 -S -D -m 2 -F 0.002 –u' with the 'mpileup' command to detect SNPs. To eliminate SNP calling errors caused by incorrect mapping or INDEls, only high-quality SNPs  were retained for further analysis. After filtering, 8,258,753 SNPs were retained from the initial set of 18,875,282 raw SNPs.After alignment, we applied the Bayesian approach to SNP calling at the population level using the Samtools software57. Genome annotation was used to classify SNPs into different categories, including exonic (overlapping with a coding exon), intronic (overlapping with an intron), splicing site (within 2 bp of a splicing junction), upstream (within 1 kb of the transcription start site), downstream (within 1 kb of the transcription stop site), and intergenic regions. SNPs in coding exons were further categorized as either synonymous (no amino acid change) or non-synonymous (amino acid change). In this case, only the highest-quality SNPs were annotated. In addition, mutations causing stop gain (premature stop codon resulting in a truncated protein) and stop loss  were also classified in this group.SNP annotation was performed using the ANNOVAR package (Version: 2013-05-20) with alignment to the sugar beet DNA reference genome (RefBeet-1.2.2)http://treesoft.sourceforge.net/treebest.shtml) for tree construction and MEGA6.0 software (http://www.megasoftware.net/) for visualization of the phylogenetic trees, which revealed the clustering patterns and underlying genetic relationships among different sugar beet accessions. To further assess the genetic structure and identify possible population subgroups among the accessions, we performed PCA using GCTA software (http://cnsgenomics.com/software/gcta/mlmassoc.html). The Tracey-Widom test was used to analyse the eigenvectors' significance level, providing statistical support for the observed patterns of genetic variation. Using fastSTRUCTURE58, a population admixture analysis with K = 2 to K = 5 was performed to infer ancestral admixture.To gain deeper insights into the phylogenetic relationship from a genome-wide perspective, we constructed an individual-based NJ tree using the p-distance metric. This approach allowed us to investigate the genetic relatedness among the sugar beet accessions. We used TreeBeST software (2) of each SNP pair in a chromosome59. An LD decay plot was generated using the average r2 value for distances from 0 to 1000 kb. Pairwise IBS calculations were also performed using PLINK, and a distance matrix was generated for each subpopulation. Population genetic diversities were measured using VCFtools60 by calculating θπ and Fst. θπ was used to measure the genetic diversity of each subpopulation, while Fst was used to measure the genetic diversity between subpopulations. In addition, sliding window calculations of r2, θπ, and Fst were performed for genome-wide representations of sugar beet genetic diversity with a 100 kb window with 10 kb steps.LD analyses for each subpopulation were performed using PLINK by calculating the correlation coefficient (r61. To minimize false positives and increase statistical power, an MLM analysis was performed by GAPIT without compression. The significance threshold of SNP-trait associations was established with a false-detection rate-adjusted P < 1 × 10–6 using the Benjamini–Hochberg procedure62. The GWAS results were displayed using a Manhattan plot and a QQ plot created with the R package CMplot63.Association analysis between target traits and genome-wide SNPs across the subpopulations from structure analysis was carried out by using the GAPIT package in R software–6 and their associated traits were retained among all localized and annotated pleiotropic genes to build the phenotype-gene network for sugar beet. PPI information for sugar beet was retrieved from the STRING database64 and mapped to the sugar beet genes using BLAST65. The construction, visualization, and exploration of the network were performed using Cytoscape66.The most significant pleiotropic genes with P < 1 × 10Supplementary Information.


Water adsorptionisotherms are systematically summarized by usingcelluloses and polysaccharides as hydrophilic crystal/amorphous materialswith functional groups, silicas as hydrophilic porous materials, andinorganic salts, sugar alcohols, and amino acids as hygroscopic deliquescentmaterials. For hydrophilic fibers such as celluloses and polysaccharides,water was adsorbed on amorphous solids, and water clusters were formedaround functional groups. For porous materials such as silicas, capillarycondensation occurred in the micropores of silicas. For deliquescentmaterials such as inorganic salts, sugar alcohols, and amino acids,water adsorption rapidly increased stepwise at a specific thresholdrelative humidity, accompanied with a structure transformation toa liquid state. In addition, the water activity (Aw) of materialsused in packed products was able to be estimated from the water adsorptionisotherms of the pure component. This indicated that the deliquescentmaterials have a great effect on the depression of Aw for the suppressionof microbial growth at an extremely high water content. The deliquescentmaterials could be useful to develop new environmentally and sustainableproducts and technologies with the mediation of water vapor and/orhydration. Differenttypes of materials exhibit different water capacities and shapes ofwater sorption isotherms. Therefore, systematic water adsorption isothermsof materials used in products are crucial to predicting the watercontent and quality of products during storage, transport, and marketing.First, product quality depends on the water content of materialsused in packed products. Water vapor is then adsorbed on each componentused in packed products and transfers outside the products throughthe packaging material based on the water adsorption isotherms andpermeability through the packaging material.8 Moist foods are liable to be contaminated by certain microorganisms,and their microbial growth requires nutrient and temperature to acertain extent as well as free water available to bacteria. Hence,FDA sets the water activity (Aw) of foods to 0.85 or less as a criterionfor preventing food spoilage and microbial growth.9 In addition, salt preservation and sugar preservation methodsare well-known to depress water activity.8 Herein,water activity is a water vapor pressure (p) in equilibriumwith materials having a given water content divided by a saturatedvapor pressure (po) of water at the sametemperature11Second, the hygroscopic characteristics of materials used in packedproducts play an important role in the product preservation and hygieneof products during storage, transport, and marketing.11 Therefore, to prevent microbial contamination and storeproducts for a long time, the water activity should be set to 0.85or less by adding inorganic salts and sugars.The water activity at a given water content can be determinedbyadsorption thermodynamics or solution thermodynamics.13 Furthermore, applying deliquescence to products from a differentpoint of view, a formulation has been filed in which an active ingredientis retained in a glassy transparent film formed by a deliquescentsugar alcohol and is gradually released in a sustained manner uponuse as a product for transmucosal oral administration of the activeingredient.14 Qualitative and quantitativeinformation regarding deliquescence may also be helpful for improvingproduct qualities and adding a new feature with an associated self-organizingsurface layer.Third, scientistshave been interested in deliquescent materials.Studies on the hygroscopic behavior of deliquescent atmospheric aerosolparticles such as NaCl have shown that an adsorbed water layer incore–shell form on the surface of airborne atmospheric aerosolslead to impacts on global climate change.The objectives of this study are to summarizethe water absorption–desorptionisotherms for various hydrophilic materials such as celluloses andpolysaccharides as hydrophilic crystal/amorphous materials with functionalgroups, silicas as hydrophilic porous materials, and inorganic salts,sugar alcohols, and amino acids as hygroscopic deliquescent materials,to discuss the hygroscopic characteristics of water adsorption behavior,and to provide useful information on the aspects of water-relatedutilizations with the mediation of water vapor and/or hydration.22.1Forcelluloses, Vitacelpowdered cellulose samples of L600-30 with a particle size of 30 μm,L00 with a particle size of 120 μm, and LC200 with a particlesize of 300 μm and a Vivapur microcrystalline cellulose sampleof 200 with a particle size of 190 μm were supplied by J. Rettenmaier(USA). Viscopearl porous cellulose was supplied by Rengo Co., Ltd.(Japan). Spherical microcrystalline cellulose (MCC) was supplied byAsahi Kasei Corporation (Japan).For polysaccharides, pectinwas purchased from Herbstreith & Fox GmbH & Co. KG Pektin-Fabriken,Neuenbürg (Germany) and gellan was purchased from San-Ei GenF.F.I., Inc. (Japan). Gum arabic, sodium alginate, κ-carrageenan,xanthan gum, tragacanth gum, and agar were purchased from FUJIFILMWako Pure Chemical Corporation (Japan). Citrus fiber was suppliedby CP Kelco (USA).For silicas, Sylopage was supplied by FujiSilysia Chemical Ltd.(Japan), Sipernat 2200, Sipernat 22, Sipernat 25, and Sipernat 33were supplied by Evonik (Germany), and Carplex BS-305, Carplex BS-XR,and Carplex DSL-67SD were supplied by Evonik (Japan). M.S.GEL wassupplied by AGC Si-Tech Co., Ltd. (Japan).For salts, sodiumdihydrogen phosphate, potassium carbonate andsodium carbonate, and sodium chloride were supplied by Univar (UK)and trisodium phosphate was supplied by Israel Chemical Limited (Israel).Calcium lactate was supplied by Taihei Chemical Industrial Co., Ltd.(Japan).For sugar alcohols, xylitol was supplied by Roquette(France) andmaltitol was supplied by Caldic Nordics (Sweden).For aminoacids, arginine was purchased from Kyowa Hakko Bio. Co.,Ltd. (Japan), γ-aminobutyric acid was purchased from Altaquimica(Spain), and monosodium glutamate was purchased from Marugo Corporation(Japan).For sweeteners, acesulfame K was supplied by Univarand sucralosewas purchased from FUJIFILM Wako Pure Chemical Corporation (Japan).2.2p/po is within 0.3% of the reading variation per 300 s, and that rangingfrom 90% R.H. to ca. 95% R.H. is within 0.3 Pa of the reading variationper 300 s.Water vaporadsorption isotherms (at 25 °C) of samples were measured volumetricallyby using an adsorption apparatus . To remove any organic residues and moisture from the samplesurface without surface reactions, the silica and others were evacuatedbelow 0.1 Pa at 250 °C for 2 h and 105 °C for 1 h beforeadsorption, respectively. The preliminary tests confirmed that thesedesorption conditions were adequate to ensure that the adsorbed watercould be measured with good reproducibility. The tolerance rangingfrom 0 to 90% R.H. 3The water adsorption isotherms of celluloses, polysaccharides,silicas, salts, sugar alcohols, amino acids, and others are shownin 3.13 The wateradsorption isotherms are nonlinear, typically sigmoidal in shape,and have been classified as type II.17 However, the water contentof celluloses was less than those of others and was not related tothe fiber length. It is reported that the water adsorption of celluloseis related to the cellulose crystallinity and pore volume.20 Although Tammelin et al.21 reported thata more crystalline cellulose film possessed nanoporosity, more surfacearea (more binding sites for water), and higher adsorption capacityof water when compared to an amorphous cellulose film, the lower watercontent of Vivapur and MCC with higher crystallinity22 has proven that they have less porosity, as shown in 24 This means that water capacity depends onthe small pore volume ranging from micropores to mesopores  along with the larger surfacearea.As shown in the water adsorptionisotherms of celluloses in Moreover, water adsorption isothermsof citrus fiber are also shownin The macropore size distribution of the sampleswas determined bymercury intrusion porosimetry using a Pore Master 60-GT ; it can cover pores ranging from6.5 nm  to 10 μm (at20 psia). The sample weight is 0.2 g. The cell size is 10φ ×30 mm, and its volume is 0.5 cc. The contact angle of mercury is 140°,and the surface tension of mercury is 480 dyn/cm. The samples wereevacuated for 105 °C and 2 h before measurement.3.21 The water adsorption isotherms of the polysaccharides are shownin 22 Polysaccharides have plenty of functionalgroups, such as a carboxyl group and amino group. Water adsorbed onthe functional groups in the first step, and a water cluster was formedaround the first adsorbed water molecules in the second step.27 Thus, it can be seen that the water content of the polysaccharidewas relatively high, and for example, at the condition of an Aw of0.85, the water content of the polysaccharide is 2 to 5 times greaterthan that of celluloses. The water adsorption isotherms of polysaccharideincreased in the following order: gellan < tragacanth gum <pectin, gum arabic, κ-carrageenan, agar < sodium alginate,xanthan gum.The potential usesof natural polysaccharides have recently attracted interest in waterpurification, drug delivery, tissue engineering, agriculture, andantimicrobial and biomedical applications.3.330 Recently, theprecipitated silica having a structure close to that of gel-type silicahas also been produced by using a synthesis method in which primaryparticles are grown in an aggregated state after causing aggregationwhile controlling the reaction temperature, pH, and salt concentrationof the synthesis of silica primary particles and suppressing the growthof primary particles.30 The gel-type silica is featuredby a high pore volume of micropores and mesopores without macropores.30 The poreproperties and silanol group numbers of precipitated silica and gel-typesilica are listed in Wet processsilica has severaltypes, such as precipitated and gel-type silica. The precipitatedor gel-type silica was synthesized by reacting a sodium silicate solutionwith sulfuric acid under an alkaline or an acid condition, respectively.The precipitated silica is produced by neutralizing silica at reactionconditions of a relatively high temperature and an alkaline pH range.The primary particles grow at a faster rate and are highly condensedin flock-like form, which are then washed with water, filtered, anddried before consolidation. Alternatively, the gel-like silica isproduced by consolidating the primary particles under the reactionconditions where the growth of the primary particles is suppressedby allowing the neutralization reaction to proceed in an acidic pHrange, and the primary particles are transformed into a three-dimensionalnetwork structure. The porous gel is reacted using a mixing nozzlehaving a strong shear, and then, the gel-like silica is produced bywashing with water, drying, and pulverizing.SBET) ofthe samples was determined from N2 adsorption–desorptionisotherms obtained at 77 K using a BELSORP-max ; the specific surface areas were calculated using the Brunauer–Emmett–Teller(BET) method. The pore volume and pore size distribution were determinedusing the Barrett–Joyner–Halenda (BJH) for mesoporesand macropores and Horvath–Kawazoe (HK) methods for micropores.40 A micropore, mesopore, and macropore are definedas a pore with a diameter of ≤2 nm, 2–50 nm, and ≥50nm, respectively. The total pore volume was assessed to be the N2 adsorption volume at a relative pressure (p/po) of 0.99. The number of silanol groups) on the surface of silicaby titration was determined by a method as developed by Sears.41 The Sears number was given asB is the consumed volumeof sodium hydroxide between pH 4 and pH 9 (mL), NA is Avogadro’s number, A is thesample weight (g), and SBET is the specificsurface areas (nm2/g).The specific surface area  andrapidly increased stepwise at the threshold relative humidity, ina similar behavior to the salts, as shown in The water adsorptionisotherms of sugar alcohols are shown in 3.636 The hygroscopicity of amino acids in atmospheric aerosols was alsoreported to be influenced by whether or not crystallization and deliquescenceoccur.37To understand the deliquescencecharacteristics of amino acids at an extremely high relative humidityranging from 80 to 99.5%, the water adsorption isotherms were measuredeven when liquefied, as will be mentioned later. The water adsorptionisotherms of amino acids are shown in 38 The findings showed that their water uptake is attributed to liquefactionover the threshold relative humidity. Both γ-aminobutyric acidand monosodium glutamate changed from white powder to viscous transparentfluid when deliquescence occurs at a high relative humidity . When evaluated using aspectrophotometer , γ-aminobutyricacid and monosodium glutamate after the water adsorption experimentsare shown as transparent. After a while, γ-aminobutyric acidremains a viscous transparent film, while monosodium glutamate becomesan opaque solid . The stability of deliquescence appears to depend on the stabilityof a quasi-stable state of the liquid state.First, the water adsorptionisotherms of γ-aminobutyric acid and monosodium glutamate exhibitedthe combined isothermal adsorption isotherms (between type II andtype IV) and increased stepwise at the threshold relative humidity.The adsorption capacity of water can reach as high as 1.7 times thesample weight for γ-aminobutyric acid and 2.5 times the sampleweight for monosodium glutamate. The threshold relative humidity is74% for γ-aminobutyric acid and 86% for monosodium glutamate.γ-Aminobutyric acid and monosodium glutamate are deliquescentsand have a high adsorption capacity of water. It is reported thata deliquescent material exhibits a solid to solution phase transformationwhen the relative humidity reaches its threshold, with kinetics increasingas the relative humidity further increases.Second, althoughthe water adsorption isotherm of arginine increasesstepwise at the threshold relative humidity of 40%, the adsorptioncapacity of water is just 0.2 times the sample weight and has a lowadsorption capacity of water.3.7As shown inthe water adsorption isotherms of sweeteners in 3.8For the suppression of microbial growth, a massfraction of mixture with an Aw of 0.85 or less at a given water contentis essential to be fixed experimentally by adjusting deliquescentmaterials such as salt, sugar, and the like. When the water activitycan be calculated by using the water adsorption isotherm from variousmass fractions, the mass fraction having the Aw of 0.85 or less canbe efficiently determined.11 reported the measurement of water sorption isotherms ofsuperabsorbent polymers, and their water activity was evaluated bysolution thermodynamics to prevent microbial contamination duringtransportation or storage. However, since it is difficult to applysolution thermodynamics to solids that do not dissolve in water, thewater activity was determined by adsorption thermodynamics. In thisstudy, the total water content of multicomponents at a given Aw isequal to a weighted average of water contents of pure components ata given Aw, as reported in a previous study.39 That is, the water activity of the mixture is calculated by summingthe multiplication of each water content by its mass fraction in thepure component.Kumagai et al.The estimation of Aw was proven as follows.First, a mass fractionof the mixture was determined in equilibrium with the Aw of 0.85 underthe condition of a water content of 50% wet basis (w.b.) by experimentallyrepeating the mixing and the measurement of Aw with a LabMaster-awNEO . Second, the water content of a purecomponent in equilibrium with the relative humidity of 85% was obtainedfrom the water adsorption isotherm, and the total water mass of themixture was calculated by summing the water mass in each component,as listed in 31 reported that synergisticinteractions, among the deliquescent components partially or completelydissolved, occur among the blending of NaCl and maltodextrins whenthe deliquescent components such as sodium chloride were in contactwith or in close proximity to the amorphous maltodextrin particles.However, as the water content of the mixture can be calculated bythe pure adsorption isotherm, the blending of the deliquescent solidsand the others has little synergistic effect. This indicated thatplenty of water adsorbs on the deliquescent solids around the Aw of0.85, except for maltitol with a threshold relative humidity of 87%and sodium glutamate with a threshold relative humidity of 86%, anda further increase in the equilibrium water content by blending doesnot occur.Ghorab etal.The water contents (% w.b.) in equilibrium with theAw of 0.85in the adsorption and desorption routes are listed in 4The water adsorption–desorption isotherms of celluloses,polysaccharides, silicas, salts, sugar alcohols, amino acids, andothers were measured. Celluloses and polysaccharides were used asthe hydrophilic crystal or amorphous materials with functional groups,silicas were used as the hydrophilic porous materials, and the inorganicsalts, sugar alcohols, and amino acids were used as the deliquescentmaterials.For the celluloses and polysaccharides, polysaccharideswith plentyof functional groups have higher adsorption capacity of water thancelluloses, with associated cluster formation. For the silicas, theadsorption capacity of water depends on their pore volume based oncapillary condensation into the micropore. The gel-like silicas thatwere synthesized under acidic conditions have a greater pore volumeof micropores and mesopores without macropores than the precipitatedsilica that was synthesized under alkaline conditions. The equilibriumwater contents of the gel-type silicas showed 2–7 times morethan those of the precipitated silicas at a relative humidity of 85%.For the hygroscopic deliquescent materials, the inorganic salts inducedthe water content uptake above the threshold relative humidity, wateris absorbed on them from 5 to 10 times their own mass, and the thresholdrelative humidity ranges from 32 to 75%, with associated structuraltransformation. Sugar alcohols and some amino acids also induced thewater content uptake and have a water adsorption capacity severaltimes their own mass, but their threshold relative humidity of over77% is relatively high compared with the inorganic salts. Maltitolwith a threshold relative humidity of 87% and monosodium glutamatewith a threshold relative humidity of 86% have little effect on depressingthe Aw below 0.85.The water activity of products can be estimatedusing the wateradsorption isotherms of pure material used in products and the effecton depressing the Aw for the suppression of microbial growth increasedin the following order: celluloses < silicas < polysaccharides< sugar alcohols < amino acids < inorganic salts. In addition,γ-aminobutyric acid and monosodium glutamate can form a self-organizingsurface layer on the product with the mediation of water vapor and/orhydration, which makes it possible to improve product qualities andadd a new feature.


Primary lateral sclerosis (PLS) is a neurodegenerative motor neuron disorder that is characterized by corticospinal and corticobulbar dysfunction. In this disease, muscle relaxants in general anesthesia should be used with extreme caution.A 67-year-old woman with a history of PLS was scheduled for laparoscopic gastrostomy due to long-term dysphagia. In the preoperative assessment, she presented a tetrapyramidal syndrome with generalized muscle weakness. A priming dose of 5 mg of rocuronium was administered and the train-of-four (TOF) ratio (T4/T1) after 60 seconds was 70% so induction was followed with fentanyl, propofol, and additional 40 mg of rocuronium. After 90 seconds when T1 was lost, the patient was intubated. During surgery, the TOF ratio increased progressively until 65%, 22 minutes after a final bolus of 10 mg of rocuronium. Prior to emergence, 150 mg of sugammadex was given and neuromuscular block reversal was evidenced with a TOF ratio > 90%.As it was decided to perform the surgery laparoscopically, general anesthesia with a neuromuscular blockade was necessary. Since it is reported that patients with motor neuron diseases show an increased sensibility to non-depolarizing muscle relaxants (NDMR), these agents should be used cautiously. Adversely to what studies document, no augmented responsiveness was shown in TOF monitoring, so the standard dose of 0.6 mg/kg of rocuronium was safely given. A final bolus of NDMR was administered after 54 minutes, demonstrating a similar pharmacokinetics profile in terms of duration of action as reported in several studies (45-70 minutes). In addition, a full and rapid neuromuscular blockade recovery with 2 mg/kg of sugammadex was seen, as previously demonstrated in a case series. Primary lateral sclerosis (PLS) is an extremely rare (incidence of 1 case in 10 million), idiopathic, nonfamilial, neurodegenerative disorder with insidious onset usually in the fifth or sixth decade that is characterized by upper motor neuron dysfunction, therefore being classified as a motor neuron disorder [Motor neuron disorders are a spectrum of diseases that include amyotrophic lateral sclerosis (ALS), which is the most common motor disease. Although there is much controversy about whether PLS and ALS are distinct pathological entities or not, the absence of clinical features of lower motor neuron involvement in PLS confirmed by neurophysiological studies can differentiate these disorders .PLS is typically slowly progressive with an average symptom duration described in several series ranging from seven to 27 years in contrast to ALS, which has a rapidly progressive course and an average life expectancy of three years. The clinical hallmarks in PLS are restricted to corticospinal and corticobulbar dysfunction so at its onset, patients can present stiffness, clumsiness, or mild weakness with signs of spasticity and hyperreflexia, dysarthria, dysphagia, and emotional lability, while limb wasting is a rare feature of PLS compared to ALS . MoreoveAs PLS represents 1-3% of all the patients with motor neuron disease, literature concerning anesthetic management in PLS patients is scarce, with no case reports published until the date of usage of neuromuscular block agents in general anesthesia in this specific disorder. Therefore, we present the first case of balanced general anesthesia in a patient with PLS undergoing laparoscopic permanent gastrostomy and discuss perioperative concerns to consider when managing a patient with this uncommon motor neuron disease.A 67-year-old woman with a history of PLS was scheduled for laparoscopic gastrostomy due to long-term dysphagia. The patient’s PLS symptoms began 11 years before surgery when the patient presented equilibrium disturbances that resulted in frequent falls. Through the years, she developed a tetrapyramidal syndrome with generalized muscle weakness, bilateral lower extremity spasticity, and hyperreflexia as well as dysarthria and dysphagia to solids and liquids.The patient’s surgical history was significant for thyroidectomy in 2004 and gastric bypass surgery in 2006, under general anesthesia without apparent anesthetic complications. The patient did not document any drug allergies, history of smoking, or alcohol consumption. The patient's medical history included parkinsonian syndrome, hypothyroidism, well-controlled arterial hypertension, urinary incontinence, and vitamin D insufficiency. The patient’s medication included baclofen, riluzole, levodopa, lansoprazole, levothyroxine, indapamide, oxybutynin, clonazepam, fluoxetine, benserazide, cholecalciferol, and calcium carbonate.2. Hemodynamically, the patient was stable with tensional values of 109/60 mmHg and other vital signs in the normal range - heart rate 65 bpm, oxygen saturation 96% at room air, and tympanic temperature 36.6ºC. The patient revealed bilateral lower extremity spasticity as well as bulbar symptoms. She presented limited functional capacity (<4 METS) due to her neurological disorder requiring wheelchair use and admitted snoring with morning sleepiness and headaches, although polysomnography did not document obstructive sleep apnea criteria. Functional pulmonary tests showed forced vital capacity (FVC), forced expiratory volume in 1s (FEV1), and ratio FEV1/FVC within the normal range. However, maximal static respiratory pressures were decreased with maximal inspiratory pressure (MIP) at 61% of predicted and maximal expiratory pressure (MEP) at 78% of predicted, therefore the patient had started noninvasive ventilation with nasal continuous positive airway pressure (CPAP) eight months prior to surgery and cough-assist two months later to facilitate mobilization of airway secretions.The preoperative assessment of the patient was done the day before the surgery where it was documented that her height was 164 cm, weight was 75 kg with a BMI of 27.89 kg/mOn the day of the surgery, the patient was premedicated with pantoprazole 40 mg and was consequently monitored following American Society of Anesthesiologists (ASA) standards  and the bispectral index (BIS) and neurostimulator. Arterial cannulation was performed in the left radial artery in order to take serial arterial blood gas tests to evaluate oxygenation status after surgery. The patient was preoxygenated for three minutes and after a priming dose of 5 mg of rocuronium was administered, a set of train-of-four (TOF) stimuli was applied over the ulnar nerve and repeated every 12 seconds in order to access patient sensibility to non-depolarizing muscle relaxant (NDMR). As the TOF ratio (T4/T1) after 45 s was 70%, induction was followed by a bolus of fentanyl 50 mcg, propofol 100 mg, and rocuronium 40 mg. After additional 90 seconds when T1 was lost, the patient was intubated with a MAC3 laryngoscope blade without any difficulty and a 7.0 endotracheal tube was placed. Mechanical ventilation was started afterward in a pressure control-volume guaranteed mode, maintaining anesthesia with sevoflurane 2% in a 50% oxygen-50% air mixture. Fifty-four minutes later, an additional bolus of 10 mg of rocuronium was given as the TOF ratio (T4/T1) was 30%. The patient remained hemodynamically stable with oxygen saturation (SpO2) >99% and the TOF ratio increased progressively until 65%, 22 minutes after the previous and final bolus of rocuronium. A schematic chronological view of the rocuronium bolus administered according to quantitative neuromuscular monitoring is represented in Figure Prior to emergence, 150 mg (2 mg/kg) of sugammadex was given and neuromuscular block reversal was evidenced with a TOF ratio >90%. The patient was promptly suctioned and extubated after responding to verbal commands and being able to open her eyes spontaneously and consequently transferred to the hospital’s post-anesthetic care unit. Postoperatively, analgesia was managed with acetaminophen 1000 mg and for nausea and vomiting prophylaxis a bolus of ondansetron 4 mg was given. The patient was discharged five days later, hemodynamically stable with SpO2 >98% with nasal CPAP. PLS is an uncommon pre-junctional disorder that affects upper motor neurons that innervate the voluntary musculature and represents a challenge to the anesthesiologist, as patients have an increased risk of pulmonary perioperative complications. In this case, we present a patient with 11-year PLS evolution with spasticity, significant bulbar symptoms, and ventilatory support at home, which increases dramatically the risk of aspiration and respiratory failure especially in upper abdominal surgery . FurtherThe decision to perform the surgery laparoscopically permits the pneumoperitoneum to distend and separate the abdominal wall from its contents enhancing easier manipulation of the anatomical structures that were distorted after gastric bypass surgery . Thus, gMoreover, it is important to consider that patients with this disorder are more sensitive to sedatives . Hence, Finally, in the postoperative period, close monitoring of vital signs especially oxygen saturation with a pulse oximeter is essential to detect respiratory depression. The eight-month history of non-invasive ventilation (NVI) and its restitution after surgery in this patient could also have a beneficial role in her respiratory function as it has been reported that NVI decreases the reintubation rate .In conclusion, anesthesiologists can encounter difficult challenges due to uncommon diseases and consequently anesthetic management should not only be addressed to the disorder specificities but also to the clinical presentation of the patient by the time of surgery, medical and surgical history, type of surgery, and more specifically in this case to the increased risk of aspiration and respiratory failure.In this patient, general anesthesia was the clear option, and we can document that standard doses of rocuronium were successfully administered, as she did not show increased sensitivity and that the pharmacokinetics profile of rocuronium in this patient was comparable to healthy patients. Also, sugammadex was safely given to recover neuromuscular blockage in the recommended dose for the TOF ratio presented at the time of reversal.


With millions of victims worldwide, multiple sclerosis is the second most common cause of disability among young adults. Although formidable advancements have been made in understanding the disease, the neurodegeneration associated with multiple sclerosis is only partially counteracted by current treatments, and effective therapy for progressive multiple sclerosis remains an unmet need. Therefore, new approaches are required to delay demyelination and the resulting disability and to restore neural function by promoting remyelination and neuronal repair.The article reviews the latest literature in this field.The fibroblast growth factor (FGF) signaling pathway is a promising target in progressive multiple sclerosis.FGF signal transduction contributes to establishing the oligodendrocyte lineage, neural stem cell proliferation and differentiation, and myelination of the central nervous system. Furthermore, FGF signaling is implicated in the control of neuroinflammation. In recent years, interventions targeting FGF, and its receptor (FGFR) have been shown to ameliorate autoimmune encephalomyelitis symptoms in multiple sclerosis animal models moderately.Here, we summarize the recent findings and investigate the role of FGF/FGFR signaling in the onset and progression, discuss the potential therapeutic advances, and offer fresh insights into managing multiple sclerosis. Fibroblast growth factors are involved in the MS development and crucial processes. FGF ligands exercise their functions by binding with their high‐affinity receptor family (FGFR1‐4) and are involved in fundamental physiological processes in adults, including wound repair, angiogenesis, and metabolism.22.1+ microgliaMultiple sclerosis is a chronic inflammatory demyelinating and degenerative condition of the CNS. Inflammation of the spinal cord and brain is invariably present in all phases of MS and declines with disease progression.2.214C methods, it was found that the demyelination is partly caused by the depletion of the myelination ability of the surviving oligodendrocytes rather than by an impairment in OPC differentiation.Demyelination leads to a reduction in axonal integrity and, over time, to neuronal dysfunction. Neurodegeneration is a characteristic of MS and a major contributor to clinical impairment and decreased quality of life. Demyelinated axons become frangible and suffer damage from activated immunological and glial cells releasing cytokines, oxidative products, and free radicals. Even in the predominantly inflammatory demyelinating stage of the disease, transected axons are abundant, demonstrating that axonal loss occurs at disease onset and continues with time. In the initial phases of RRMS, the axonal loss has no immediate substantial clinical impact. As lesions accumulate with time, however, the clinical aspects of MS become driven by axonal loss. Thus, it is believed that the brain's ability to adjust for further axonal loss exhausts before RRMS and SPMS shift. At this stage, MS lesions include remyelination, inflammation resolution without repair, or a “smoldering” state of coexistence of inflammation and myelin degeneration.2.3Demyelination can occur parallel to regeneration processes, which restore some of the destroyed myelin‐generating cells and rebuild the myelin sheath around axons. This is accomplished by the activation, migration, and polarization of resident OPCs and neural stem cells (NSCs) into myelin cells, initiating an oligodendrocyte‐driven repair process known as remyelination. The identity of cells that cause remyelination in the CNS of MS patients has been a subject of debate. OPCs and mature oligodendrocytes that have survived are two potential candidates. This discussion is crucial because therapeutic approaches to enhance remyelination may differ depending on the specific cellular pathways involved. Lineage tracing experiments revealed that newly generated oligodendrocytes derived from OPCs form new myelin sheaths in demyelinated regions.33.1The 18 FGFs cluster into six subfamilies, with the FGF1, FGF4, FGF7, FGF8, and FGF9 subfamilies functioning in a paracrine manner and the FGF19 subfamily members operating as endocrine factors.3.23K)/protein kinase B (AKT) signal; and mitogen‐activated protein kinase (MAPK) signal.3), and MKP1 can also negatively regulate FGF/FGFR signaling to varying extents. These regulatory factors modulate intracellular signaling or ligand bindingUpon ligand binding, FGFRs dimerize and cause the phosphorylation and activation of intracellular tyrosine kinases.4FGF/FGFR signaling not only plays a crucial role in CNS formation during development but also has a broad function in the adult CNS. A prototype member, FGF1, is expressed by neurons in adult neural tissue and acts as a mitogen in neurodevelopmental processes.The CNS myelin‐producing cells, oligodendrocytes, play a central role in generating and preserving the pace and the power of axonal electrical impulses. Neurological deficits in MS result from myelin damage or insufficient remyelination. Understanding the signals involved in developing oligodendrocyte‐driven myelination may shed light on demyelinating disease prevention and treatment. OPCs migrate to various brain regions during development, transforming into myelin‐producing cells.3K/AKT/mTOR, the downstream mediators of these FGFR signaling pathways, sequentially regulate myelination by affecting distinct stages of the oligodendrocyte lineage. ERK1/2 signaling regulates both the transition from early to late OPCs and subsequent immature oligodendrocyte stages. The mTOR signaling pathway is necessary to transform from immature to mature oligodendrocytes.Furthermore, ERK1/2 and PIImmunity is continually regulated by FGF signaling, which is, in turn, modified by immune cells during inflammation and tissue healing. FGF signaling cascades are important in CNS inflammatory responses by modulating immune homeostasis and host defense. FGF1 reduces the inflammatory response associated with neuropathic pain by inducing the production of the Th2 cytokine IL‐4, upregulating arginase‐I (Arg‐I), and suppressing the activation of microglia and astrocytes.55.1In MS, FGF1 is predominantly expressed in remyelinated lesions, with its production being lower in the demyelinated lesion core than in the remyelinated rim.−/− animals. The role of FGF2 is variable since chronically high FGF2 levels in the CSF reverse the positive benefits, and the secretion of FGF2 by immune cells increases with disease progression in MS/EAE.How FGF2 affects oligodendrocyte responses during demyelination and remyelination in MS is debatable. On one hand, FGF2 is regarded as a neuroprotective agent that promotes remyelination in MS.5.2FGF8, FGF17, and FGF18 belong to the FGF8 subfamily. They share 60 to 80 percent of amino acid sequence similarity and have comparable receptor binding features.5.3The cascade of pathogenic events in MS eventually leads to the loss of neurons and axons, which can be measured by reduced brain volume on volumetric magnetic resonance imaging (MRI) in vivo. Between 0.5% and 1.5% of MS patients develop brain atrophy each year, and during the progressive stages of the disease, the deep gray matter structures display a higher rate of degeneration.5.4The endocrine FGF subclass ligands  are also known as the FGF19 subfamily. FGF21, mainly released by the skeletal muscle, pancreas, liver, kidney, and adipose tissue, exerts pleiotropic effects in regulating glucose, lipid, and energy homeostasis.2D3 levels.2D3 has been reported to have the opposite effect on cytokine secretion of macrophages.2D3 on immune responses. Accordingly, it is conceivable that elevated FGF23 levels contribute to MS by promoting inflammation.FGF23 was first identified in the ventrolateral thalamic nucleus of the mouse brain, and its physiological role has recently attracted significant attention.5.5ind−/− and FGFR2ind−/− animals, myelin, axonal loss, and the infiltration of inflammatory cells were decreased in the chronic phase of EAE without causing any alterations in the acute phase. The protective effects of the oligodendrocyte‐specific deletion of FGFR1 and FGFR2 on EAE are mainly manifested in two aspects. First, the knockdown of FGFRs can regulate the inflammatory environment in the CNS. T cells and B cells, as well as macrophages and activated microglia, were significantly reduced in the spinal cord and cerebellum of FGFR1ind−/− mice, along with the decreased expression of the proinflammatory cytokines TNFα, IL‐1β, and IL‐6, and the chemokine CX3Cl1 and its receptor CXC3R1.ind−/− mice showed a similar phenotype,ind−/− mice varied slightly, with an increase in PLP‐positive cells in the spinal cord and a decrease in the expression of SEMA3A. At the same time, there were no significant increases in pERK or TrkB,To characterize the role of FGFRs in oligodendrocytes in vitro, researchers used the multi‐kinase inhibitor and FGFR1‐3 inhibitor to block FGFR signaling. The application of inhibitors increased the expression of the myelin‐associated proteins PLP and 2′,3′‐cyclic‐nucleotide 3′‐phosphodiesterase. Furthermore, upregulation of neurotrophic factor BDNF/TrkB signaling and decreased expression of the remyelination inhibitor semaphorin 3A were observed. These effects may be related to the reduction in downstream molecules of the FGF/FGFR signaling pathway, such as ERK and AKT phosphorylation, in oligodendrocytes.6Current MS treatments have achieved near complete control over RRMS and focal neuroinflammation. However, no effective treatment strategies are available concerning neurodegeneration and disability. FGF signal transduction is involved in the development of the CNS, enhances the proliferation and axonal growth of NSCs in adults, and promotes their differentiation into various CNS cells, such as neurons and oligodendrocytes. Furthermore, the development of oligodendrocytes is inseparably linked to FGFs, and distinct FGFRs are expressed at different stages of their development; therefore, FGFs play pivotal roles at each stage. Surprisingly, several studies have shown that neuroinflammation is regulated partly by FGF signaling, as FGF1, FGF2, and FGF21 have anti‐inflammatory functions, while FGF9 induces neuroinflammation. These results all emphasize the importance of FGF/FGFR signaling in the pathogenesis of MS. Some FGFs are mainly expressed in acute demyelinating lesions and mediate disease progression. Others display high concentrations in remyelinating lesions in the chronic phase of MS and promote the life cycle of OPCs and the maturation of oligodendrocytes, which helps to enhance tissue repair and restore motor function .The authors declare that they have no competing interests.


ARHGEF10, PCDH7, CST6, and ROBO3) were identified, and mRNA expression and protein levels of PCDH7 in clinical samples from patients with sarcopenia, with osteoporosis, and with both sarcopenia and osteoporosis were found to be significantly higher than those from patients without sarcopenia or osteoporosis. PCDH7 seems to be a key gene related to the development of both sarcopenia and osteoporosis.Sarcopenia and osteoporosis, two degenerative diseases in older patients, have become severe health problems in aging societies. Muscles and bones, the most important components of the motor system, are derived from mesodermal and ectodermal mesenchymal stem cells. The adjacent anatomical relationship between them provides the basic conditions for mechanical and chemical signals, which may contribute to the co-occurrence of sarcopenia and osteoporosis. Identifying the potential common crosstalk genes between them may provide new insights for preventing and treating their development. In this study, DEG analysis, WGCNA, and machine learning algorithms were used to identify the key crosstalk genes of sarcopenia and osteoporosis; this was then validated using independent datasets and clinical samples. Finally, four crosstalk genes ( In recent years, sarcopenia and osteoporosis, two degenerative diseases in the motor system of older adults, have attracted increasing attention from researchers due to increasingly aging societies . For insSarcopenia is an aging syndrome characterized by decreased skeletal muscle mass, function, and strength . The firOsteoporosis is a disease characterized by low bone mass and deterioration of the microstructure of bone tissue . Its diaMuscle and bone are the most important components of the motor system, and patients with both muscle and bone disorders may face worse health and a higher risk of falls and fractures . OsteosaMuscle and bone are derived from mesodermal and ectodermal mesenchymal stem cells, with the adjacent anatomical relationship between muscle and bone providing the basic conditions for mechanical and chemical signals . The intWith the development of microarray technology and high throughput sequencing, more and more data for comparing differentially expressed genes between individuals with different diseases state have become available . Bioinfohttp://www.ncbi.nlm.nih.gov/geo/). GEO is one of the largest public databases to include microarray data and high-throughput gene expression data submitted by research institutions around the world (All datasets included in our study were downloaded and collected from the Gene Expression Omnibus (GEO) database  and pheatmap packages (version 1.0.12) in Sangerbox were used to visualize the DEGs.Sangerbox, an online tool based on R, was used to perform the DEG analysis of sarcopenia and osteoporosis datasets . GSE11102 > 0.85. Based on the soft-thresholding power calculated previously, co-expression genes were clustered and modulized and the minimum number of module genes was set at 30. The modules with a correlation of more than 80% in sarcopenia and more than 70% in osteoporosis were merged for the second time. Finally, the module eigengene and the correlation between the module eigengenes and clinical data were visualized, and all the module genes with significant correlations with sarcopenia and osteoporosis were identified and exported for further analysis.WGCNA as a novel developed bioinformatic approach aims to find co-expressed gene modules and explore the association between gene networks and phenotypes of interest, as well as a network's core genes . Age andAfter separately identifying DEGs and WGCNA genes of sarcopenia and osteoporosis, these were imported into R software to obtain the intersection. The intersection of the DEGs of sarcopenia and osteoporosis was identified as the common DEGs, and the common WGCNA genes were defined as the intersection of WGCNA genes of sarcopenia and osteoporosis. Finally, the intersection of common DEGs and common WGCNA were collected as the crosstalk genes of sarcopenia and osteoporosis.Functional enrichment analysis was performed to explore the biological functions and pathways involved in the genes we identified. Biological process (BP), cellular component (CC), and molecular function (MF) from Gene Ontology (GO) were used to describe gene information, functions, and interaction, and the pathway results were obtained from the Kyoto Encyclopedia of Genes and Genomes (KEGG) databases . R packahttps://string-db.org/) to construct PPI networks with a confidence of more than 0.7 (The protein–protein interaction (PPI) network may reveal the protein interactive relationships. The common DEGs, common WGCNA genes, and crosstalk genes were uploaded to STRING  regression were conducted based on R packages “xgboost” and “glmnet.” In the XGBoost analysis, the importance of each gene was viewed and collected. The coefficient of punishment (λ) was identified according to the AUC of the modules with different λ, and then the predictive value of genes was exported from the modules with the most suitable λ. The intersection of genes screened by XGBoost and LASSO regression was identified as the key crosstalk genes.t-test was used to compare the expression between groups, and all p-values < 0.05 were considered statistically significant.The expression of key crosstalk genes from GSE167186 and GSE35956 datasets were collected to verify the key crosstalk genes. The All participants were recruited from patients awaiting hip arthroplasty due to hip fractures who were admitted to the Department of Traumatology, Shanghai East Hospital, Tongji University, School of Medicine, Shanghai, China. The study was approved by the Institutional Ethics Committee of East Hospital ([2022] Research Review (No. 278), 7 September 2022) and was performed in accordance with the Declaration of Helsinki. The mean age of patients enrolled in this study was 65 years. Finally, 12 patients who underwent hip arthroplasty were included in our study and divided into four groups according to whether they had sarcopenia and osteoporosis. The diagnosis of sarcopenia and osteoporosis followed the guidelines mentioned previously . Skeleta® Premix Ex Taq™ . Data were normalized to GAPDH mRNA and analyzed according to the 2−ΔΔCT method. The gene primers are: forward primer TTGTGGGAGCAGGAGACAAC and reverse primer ACTCTACGAAATGGCTGTTTGC for PCDH7; forward primer GAAAGCCTGCCGGTGACTAA and reverse primer GCCCAATACGACCAAATCAGAG for GAPDH.RNA isolation was conducted following the manufacturer’s instructions for RNAeasy™ Animal RNA Isolation Kit with Spin Column . cDNA was synthesized following the manufacturer’s instructions for PrimeScript II 1st Strand cDNA Synthesis Kit . RNA quantity and integrity were measured with a NanoDrop 1000 (Thermo Scientific). The reaction system for Real-time Quantitative PCR followed the manufacturer’s instructions for SYBRProteins were extracted from tissue homogenate, and the protein concentration was quantified using a BCA kit . Total proteins of skeletal muscle and bone were subjected to 12% SDS-PAGE electrophoresis and were then wet migrated to the PVDF membrane. The membrane was sealed with 5% skimmed milk at 37°C for 2 h and then incubated with primary antibody  at 4°C overnight. The membrane was then washed with TBST three times, 15 min each time, and then cultivated with secondary antibody Goat anti-rabbit IgG with 1:2500 dilution at 25°C for 1 h. Finally, the signal was detected on the Azure imaging biosystem C300 using ECL Kit (Kangweishiji Biotechnology).https://cibersortx.stanford.edu/) was used for immune infiltration analysis and then the immune infiltration matrix generated from CIBERSORTx was imported into R software for visualization using the R package “ggplot2.” The differences between the two groups were compared using the Wilcoxon test and the correlation between the 22 infiltrating immune cells and key genes was then visualized.Immune infiltration analysis was performed to identify the roles of the immune system in the bone and muscle system. The online platform CIBERSORTx  and R software version 4.1.1 .Continuous variables were presented as mean ± standard deviation and were evaluated by independent Student’s t-test for normally distributed variables and by Wilcoxon rank-sum test for non-normally distributed data. All In this study, GSE111017 and GSE167186 were used as the datasets of sarcopenia, and GSE56815 and GSE35956 as the datasets of osteoporosis. GSE111017 was a dataset containing three sub-datasets  of data from a multi-center study in Hertfordshire, Jamaica, and Singapore. All the sub-datasets in GSE111017 were expression profiled by high-throughput sequencing using the same platforms, and the batch effect between sub-datasets was evaluated. From the evaluation results of the batch effect, there was no significant difference between the datasets . GSE1671Based on the criteria that we set for p-value and fold change of DEGs, 1,426 upregulated DEGs and 470 downregulated DEGs in sarcopenia and 1,482 upregulated DEGs and 879 downregulated DEGs in osteoporosis were identified. As shown in GO enrichment and KEGG enrichment analysis were conducted to explore the biological features of common DEGs. From the enrichment analysis visualized in 2 = 0.90. WGCNA analysis was performed to identify the gene modules of sarcopenia and osteoporosis. As shown in Biological features of common WGCNA genes were summarized and analyzed by GO and KEGG analysis. As shown in Finally, crosstalk genes were identified by intersecting the common DEGs and WGCNA genes. As shown in https://string-db.org/) to construct PPI networks and then imported to Cytoscape for visualization (COL1A1), P21 (RAC1) activated kinase 1 (PAK1), cytochrome C oxidase subunit 5A (COX5A), and cytochrome C oxidase subunit 6A2 (COX6A2); the top five genes in PPI networks of common WGCNA were fibronectin 1 (FN1), COL1A1, collagen type I alpha 2 chain (COL1A2), matrix metallopeptidase 2 (MMP2), and collagen type V alpha 1 chain (COL5A1); in crosstalk, the genes were COL1A1, aggrecan (ACAN), fibromodulin (FMOD), netrin 1 (NTN1), and asporin (ASPN).The common DEGs, common WGCNA genes, and crosstalk genes were uploaded to STRING (lization . Genes wARHGEF10), Protocadherin 7 (PCDH7), cystatin 6 (CST6), and roundabout guidance receptor 3 (ROBO3).Two kinds of machine learning models were established to identify the key crosstalk genes of sarcopenia and osteoporosis. In LASSO analysis, the change trends of gene coefficients in different λ in sarcopenia and osteoporosis are shown in GSE167186 and GSE35956 were used as the datasets of sarcopenia and osteoporosis to validate the key crosstalk genes. The expression level of four key crosstalk genes was compared between patients with and without osteoporosis or sarcopenia. As shown in To explore the relationships between the immune system and the co-occurrence of sarcopenia and osteoporosis, immune infiltration analysis was performed on the sarcopenia and osteoporosis datasets. ARHGEF10, PCDH7, CST6, and ROBO3) were identified by DEG analysis, WGCNA, and machine learning methods, and then validated by independent datasets and clinical samples. Finally, PCDH7 was identified and validated as the gene that may be significantly related to the co-occurrence of sarcopenia and osteoporosis.Sarcopenia and osteoporosis as degenerative diseases in the most significant part of the motor system have become severe health problems in aging societies. In this study, datasets of sarcopenia and osteoporosis were included to identify their possible crosstalk genes. The specimens of osteoporosis were blood monocytes and of sarcopenia were muscle biopsies. Monocytes as a type of circulation blood cells have been proven in many studies to relate to sarcopenia and osteoporosis . ConsistThe pathogenesis behind sarcopenia is still a controversial topic and is hotly debated. It is widely believed that the skeletal muscle mass in humans peaks at age 30 and then decreases significantly with age, while, in individuals aged over 75, skeletal muscle accounts for only a quarter of total body weight . MoreoveMuRF-1 and MAFbx, by regulating the TNF receptor-associated factor 6 (TRAF6) and activating the nuclear factor-kappa B (NF-κB) signaling pathway , PCDH7 was identified and validated as relating the development of both sarcopenia and osteoporosis. This may provide new insights for treating older patients with sarcopenia and osteoporosis.Of four key crosstalk genes (