We conducted a metataxonomic analysis to compare the taxonomic composition and functional profile of the fecal microbiota in healthy and diarrheic calves across three clinical periods, aiming to gain a better understanding of the changes. The findings here might provide information about pathogenic microbiota in diarrheic calves, which were found to be more enriched than in healthy calves.p = 0.074 and Shannon, p = 0.055) and beta  diversity at 1 day post-inoculation. Differences in the bacterial taxa between healthy and rota-infected calves were detected using a linear discriminant analysis effect size . Rota calves had a higher abundance of certain bacterial taxa, such as Enterococcus, Streptococcus, and Escherichia-Shigella, and a lower abundance of bacteria that contribute to the production of short-chain fatty acids, such as Alistipes, Faecalibacterium, Pseudoflavonifractor, Subdoligranulum, Alloprevotella, Butyricicoccus, and Ruminococcus, compared to the healthy calves. The observed changes in the fecal microbiota of the rota-infected group compared to the healthy group indicated potential dysbiosis. This was further supported by significant differences in the predicted functional metagenomic profiles of these microbial communities. We suggest that calves infected with bovine rotavirus had bacterial dysbiosis, which was characterized by lower diversity and fewer observed genera than the fecal microbiota of healthy calves.The objective of the present study was to identify changes in fecal microbiota and predict the functional features of healthy calves and those infected with rotavirus over time. Six Holstein calves  were randomly selected and distributed into two groups which contained three calves each. Fecal samples were taken 3 days before inoculation and on days 1 and 7 post-inoculation. The 16S rRNA gene amplicon sequencing was performed. Bacterial diversity tended to decrease in the rota group, as indicated by the alpha (evenness,  Calf diarrhea is a common problem in dairy calves and causes significant economic losses to cattle producers. Moreover, calf diarrhea can have significant long-term effects, such as delayed first calving, high rates of morbidity and mortality, growth impairment, and increased treatment costs . NeonataImproving the development and composition of the gut microbiota is one of the most important strategies for reducing the prevalence of enteric infections. Many studies have highlighted the important role of gut microbiota in animal productivity and health, including the immune system . In addiAll experimental protocols were approved by the Animal Care and Use Committee  of Sunchon National University . All the experiments were performed in accordance with the guidelines and regulations set by the governing body. Throughout the study, the health of the calves was closely monitored, including regular assessments of body temperature, body condition score, and any signs of discomfort or illness.4 TCID/mL to induce diarrhea. Following the inoculation, the consistency of calf feces was monitored day to day. Fecal consistency was scored using a 4-level scoring scale, as suggested by Renaud et al. [A cohort of six male Holstein calves from one dairy farm in Jeollanamdo province was used in the study. Holstein calves  were randomly selected and distributed into two groups, each containing three calves. The healthy (control) group consisted of three healthy calves, whereas the rota group consisted of three calves with diarrhea. The calves in the rota group were orally inoculated with 30 mL of bovine rotavirus cell culture supernatant containing a viral titer of 10d et al. , with 0 v/v) by using Ribospin™ vRD II  following the manufacturer’s instructions.Fecal samples were taken 3 days before inoculation (D − 3) and on days 1 (D + 1), and 7 (D + 7) post-inoculation. All fecal samples were collected directly from the rectum using sterile gloves and stored at −80 °C until DNA and RNA extraction. Fecal DNA was extracted using the QIAamp PowerFecal Pro DNA Kit , while RNA extraction was done from fecal suspensions  to check for rotavirus, following the test procedure provided by the manufacturer. The presence of Group A rotavirus was detected in fecal samples using qRT-PCR. The primers and probes specific for VP6 were developed by Tsuchiaka et al. . The folTo assess the quality and quantity of the extracted DNA, a NanoDrop ND-2000 spectrophotometer  was utilized. Amplicons were generated to target the V3-V4 region of the 16S rRNA gene and subsequently subjected to sequencing at Macrogen . Briefly, the 16S rRNA gene amplicon libraries encompassing both bacteria and archaea were amplified using universal primers 341F (5′-CCTACGGGNGGCWGCAG-3′) and 805R (5′-GACTACHVGGGTATCTAATCC-3′) as described by Herlemann et al. . Each DNt-test. Abnormally distributed data were further analyzed using the non-parametric Wilcoxon rank-sum test. Principal coordinate analysis (PCoA) utilized the obtained distance matrices as inputs, while the significance of sample clustering was assessed through permutational multivariate analysis of variance (PERMANOVA) employing 9999 permutations. For the analysis of differential relative abundances in the fecal microbiota and its predicted metabolic categories, we employed linear discriminant analysis effect size (LEfSe). Significance was determined based on LDA scores exceeding 2.0 and a p value threshold of less than 0.05 [p < 0.05, and a tendency of difference was declared at 0.05 < p ≤ 0.10.To assess the normality of the data, we conducted Shapiro–Wilk test using SAS 9.4 . Normally distributed data were analyzed using Student’s han 0.05 . For allAs shown in We performed 16S rRNA gene amplicon sequencing of fecal samples collected per rectum at three time points: 3 days before inoculation, one day post-inoculation, and 7 days post-inoculation, to determine the temporal changes in the calf gut microbiota in response to the rota treatment. A total of 2,157,400 amplicon sequences were generated from fecal samples from the healthy and rotavirus-infected groups. After applying a quality filter (Q score > 25) using QIIME2, we obtained 1,161,065 sequences, with an average of 64,503 ± 14,358 sequences per sample. For all samples, the calculated Good’s coverage was above 99%. In this study, the detected bacterial taxa were defined as those with a relative abundance above 0.1% and a presence in more than 50% of the animals per group.p = 0.074) and Shannon’s index (p = 0.055) tended to be higher in the healthy group than in the rotavirus group, whereas other measurements did differ. There was no difference in the alpha diversity measurements between the healthy and rota groups on D − 3 and D + 7. With regard to beta diversity analyzed using PERMANOVA, the Bray–Curtis dissimilarity in the rota group (p = 0.099) tended to differ from that healthy group on D + 1, whereas no differences was observed for D − 3 and D + 7 , Oscillospiraceae , and Rikenellaceae  was higher in the healthy group than in the rota group. At the genus level, the relative abundance of Faecalibacterium , Subdoligranulum , Pseudoflavonifractor , and Alistipes  tended to be higher in the healthy group than rota group.The taxonomic distributions of the fecal microbiota at the phylum and genus levels are shown in p = 0.064) tended to be more abundant in the healthy than rota group. At the family and genus levels, three families and two genera differed between the healthy and rotavirus groups. Briefly, at the family level, the relative abundance of Butyricicoccaceae  and Prevotellaceae  was higher in the healthy group than in the rota group. At the genus level, the relative abundance of Butyricicoccus  and Alloprevotella  was higher in the healthy group than in the rota group.On day 1 post-inoculation with bovine rotavirus, 9 phyla, 20 families, and 25 genera were identified . At the p = 0.081) and Enterococcus  were higher in the rotavirus group than in the healthy group.On day 7 post-inoculation with bovine rotavirus, 7 phyla, 13 families, and 16 genera were identified . At the Venn diagrams were used to illustrate the microbial genera shared by, or exclusively detected in, the healthy and rotavirus groups . On D − Alistipes, Faecalibacterium, Pseudoflavonifractor, and Subdoligranulum on D − 3 and Alloprevotella, Butyricicoccus, and Ruminococcus on D + 1. In contrast, Enterococcus was enriched in the rota group compared to the healthy group on D + 7.We identified potential taxonomic biomarkers in the fecal microbiota of the calves using LEfSe analysis . CompareTo predict functional biomarkers in the fecal microbiota of the healthy and rotavirus-infected groups, we performed PICRUSt2 analyses . KEGG orThe present study was designed to investigate changes in the fecal microbiota of Holstein calves, following rotavirus challenge, while minimizing the effects of external factors. Male calves were selected for each group based on age and body weight to ensure high nutritional and physiological homogeneity. The study participants were maintained under the same living conditions and fed the same diet throughout the experimental period, with no significant alterations in their diet. The primary source of nutrition for calves during the study was milk replacement. These measures were taken to control for potential confounding variables and to enable a more accurate investigation of the effects of other factors on the fecal microbiota of calves. Despite controlling for external factors, subtle differences in gut microbiota composition may have existed at baseline, which was not detected by the study design or fecal microbiota analysis. Additionally, the rotavirus challenge may have indirectly affected the gut microbiota prior to D + 1 infection through an immune response or other mechanisms.On day 1 post-inoculation with bovine rotavirus, we found that bacterial diversity decreased in the rota group via alpha (evenness and Shannon’s index) and beta (Bray–Curtis dissimilarity) diversity, which are well-known characteristics of dysbiosis . This reAlistipes, Faecalbacterium, Pseudoflavonifractor, Subdoligranulum, Alloprevotella, Butyricicoccus, and Ruminococcus; most of these were also found in previous studies [Alistipes and Alloprevotella, which belong to the Bacteroidetes, contribute to the production of short-chain fatty acids (SCFA) and further benefit gut health [Faecalbacterium and Pseudoflavonifractor are beneficial for the immune homeostasis of calves, and are positively correlated with bovine IgG [Faecalibacterium is associated with less diarrhea in neonatal calves, and this genus can produce butyrate in the gut [Subdoligranulum, Butyrcicocus, and Ruminococcus, were also enriched in the healthy group, which may act as probiotic features that positively influence the gut health of young calves, as suggested by Xin et al. [Butyricicoccus can strengthen the epithelial barrier function or defend the gut from pathogens in neonatal calves [Bifidobacterium was high, and its relative abundance in the healthy group, by more than 8% (8.53% vs. 0.03%) on D + 1. This genus has primary health-promoting functions in calves, with preventive and protective effects against diarrhea and intestinal infections [Enterococcus in the Day + 7 rota group suggests that the composition of the fecal microbiome may have been affected by rotavirus infection over time, emphasizing the potential long-term impact of rotavirus infection on the microbiome. It is possible that the immune response to rotavirus infection or other indirect effects of the infection could have altered the fecal microbiota, leading to differences in Enterococcus abundance [Streptococcus and Escherichia-Shigella in the rota group, which have been reported as opportunistic pathogens that can cause infections in immunocompromised hosts, may indicate an increased risk of gut infections and inflammation in calves [Enterococcus and Streptococcus abundance have any clinical implications for the health of Holstein calves.Most of the bacterial genera were enriched in the healthy group compared to the rotavirus group. The most important commensal bacterial genera in the healthy group were  studies ,27. Alist health . Faecalb the gut . Butyratn et al. . In partl calves . We specl calves . Althougfections . The higbundance . These fn calves ,34,35. Fn calves . These fBifidobacterium, are enriched in genes involved in folate biosynthesis, which has been verified in previous studies [Bifidobacterium. Nonetheless, folate may also play a role in promoting immune function and metabolism [Escherichia genus, which can produce a barrier effect against enteropathogens [Interestingly, we found that some of the predicted metabolic functions of the fecal microbiota were altered between the healthy and rota groups. In the healthy group, bacterial chemotaxis and secretion systems, quorum sensing, and amino acid biosynthesis were enriched. Bacterial chemotaxis and secretion systems are known as bacterial colonization and proliferation pathways, which are required for substrate transport and utilization and are ultimately crucial for energy extraction and provision . Bacteri studies ,41. Howetabolism . It is atabolism . Notablytabolism . Thus, oathogens . Howeverathogens . NeverthHowever, it is important to note that the present study used a limited number of animals, and as such, its results should be interpreted with caution. Despite the limited number of calves used in this study, our research provides valuable insights into the temporal changes in the fecal microbiota of Holstein calves following rotavirus infection. Sequential sampling at 3 days before inoculation, 1 day post-inoculation, and 7 days post-inoculation allowed us to observe the step-by-step impact of rotavirus on the fecal microbiota and gain insights into the dynamics of this infection.Enterococcus, Streptococcus, and Escherichia-Shigella, and a lower abundance of bacteria that contributed to the production of SCFA, such as Alistipes, Faecalibacterium, Pseudoflavonifractor, Subdoligranulum, Alloprevotella, Butyricicoccus, and Ruminococcus than the healthy group. Moreover, functional analysis revealed differences in metabolic pathways between the two groups, such as the enrichment of pathways related to amino acids, cofactors, and vitamin metabolism in the rotavirus group. These findings suggest that bovine rotavirus can alter the fecal microbiota composition and functional potential in calves, potentially impacting their overall health and well-being. A major goal of the dairy production system is to optimize diarrhea prevention without relying on antibiotics. Therefore, our results, which demonstrate that rotavirus infection alters the structure of the fecal microbiota and is linked to changes in metabolic functions, not only provide new insight into the treatment of rotavirus-mediated diarrhea in calves but may also be beneficial to the dairy production.In the present study, we observed differences in fecal microbiota composition and predicted functional metagenome profiles between healthy and rotavirus-infected calves at three stages: 3 days before inoculation, 1 day post-inoculation, and 7 days post-inoculation. We found that the rota group had a higher abundance of certain bacterial taxa, such as


Pasteurella multocida is a major pathogen of bovine respiratory disease, which is resistant to many of the commonly used antibiotics. We found that enrofloxacin has shown a high drug resistance in clinical treatment of Pasteurella multocida infection. In order to better understand the resistance mechanism of Pasteurella multocida to enrofloxacin, we isolated PmS and PmR strains with the same PFGE typing in vitro, and artificially induced PmR to obtain the highly resistant phenotype, PmHR. Then, transcriptome sequencing of clinically isolated sensitive strains, resistant and highly drug-resistant strains, treated with enrofloxacin at sub-inhibitory concentrations, were performed. The satP gene, of which the expression changed significantly with the increase in drug resistance, was screened. In order to further confirm the function of this gene, we constructed the deleted and complemented strains of satP, and further analyzed the function of the satP gene. After satP gene deletion, the resistance of Pasteurella multocida was obviously lower than that of wild-type strains in vitro, and the pathogenicity of Pasteurella multocida was reduced by about 400 times. We found that the satP gene is related to the tolerance and pathogenicity of Pasteurella multocida, and can be used as a target of enrofloxacin synergistic effect.Pasteurella multocida (Pm) is one of the major pathogens of bovine respiratory disease (BRD), which can develop drug resistance to many of the commonly used antibiotics. Our earlier research group found that with clinical use of enrofloxacin, Pm was more likely to develop drug resistance to enrofloxacin. In order to better understand the resistance mechanism of Pm to enrofloxacin, we isolated PmS and PmR strains with the same PFGE typing in vitro, and artificially induced PmR to obtain the highly resistant phenotype, PmHR. Then transcriptome sequencing of clinically isolated sensitive strains, resistant and highly drug-resistant strains, treated with enrofloxacin at sub-inhibitory concentrations, were performed. The satP gene, of which the expression changed significantly with the increase in drug resistance, was screened. In order to further confirm the function of this gene, we constructed a satP deletion (ΔPm) strain using suicide vector plasmid pRE112, and constructed the C-Pm strain using pBBR1-MCS, and further analyzed the function of the satP gene. Through a continuously induced resistance test, it was found that the resistance rate of ΔPm was obviously lower than that of Pm in vitro. MDK99, agar diffusion and mutation frequency experiments showed significantly lower tolerance of ΔPm than the wild-type strains. The pathogenicity of ΔPm and Pm was measured by an acute pathogenicity test in mice, and it was found that the pathogenicity of ΔPm was reduced by about 400 times. Therefore, this study found that the satP gene was related to the tolerance and pathogenicity of Pm, and may be used as a target of enrofloxacin synergistic effect. Pasteurella multocida (Pm) is a Gram-negative bacterium implicated in multi-host infections. Strains of their species are currently classified into type A, type B, type D, type E and type F serogroups [Pm is one of the primary pathogens of bovine respiratory disease (BRD) in Asia, Middle East, Africa and Europe [Pm leads to tremendous economic losses for the cattle industry due to increased mortalities and high treatment costs [Pm resistance [rogroups ,2. Pm isd Europe ,4,5. At nt costs . The trent costs ,8. Howevnt costs ,10,11,12sistance .Pm infections in animals [Pm infection in animals [Pm has recently been reported to possess acquired fluoroquinolone resistance mechanisms [Pm, gyrA gene mutation often leads to the structural change of DNA helicase, which leads to drug resistance. Within the gyrA gene, quinolone-resistant determining region (QRDR) is closely associated with drug resistance [Mycobacterium tuberculosis, Ala67–Gln106 in Escherichia coli and Arg88Ile in Pm. Detection of QRDR gene mutation has been a hot issue in the early study of fluoroquinolone resistance [Pm is extremely resistant to fluoroquinolones in vivo and in vitro, and the mechanism is mainly mediated through target mutation in gyrA and parC [Fluoroquinolones are one of the main therapeutic drugs for  animals . Enroflo animals ,15,16. Uchanisms . Among tchanisms . Mutatiosistance . Mutatiosistance . Our preand parC . The varPm which can discharge drugs, resulting in drug resistance [Pm is mainly related to the survival–nodulation–cell division (RND) family, which includes the transporters of calcium, cobalt and nickel [The bacterial cell membrane is a highly selective permeable barrier. There are some special proteins in the outer membrane of bacteria, which are called outer membrane proteins (OMPs). OMPs account for about half of the cell wall in Gram-negative bacteria. The OMPs have the functions of antibiotic, iron transport, host adhesion and maintenance of membrane integrity . At pressistance . The effd nickel . Among Gd nickel . recA, are essential for increased tolerance to antibiotic treatment by enhancing repair of DNA damage that occurs directly from antibiotic-induced DNA damage. Relevant studies have confirmed that amino acid mutations are related to the generation of tolerance [Pm [Pm tolerance to fluoroquinolones and the main targets for mediating tolerance have not been reported. To determine whether the resistance mechanism of Pm is caused by tolerance, it is crucial to work on a new tolerance mechanism and identify targets that can inhibit drug resistance.However, the development of antimicrobial resistance is a complicated multifactorial process, and related studies have shown that the formation of drug resistance also depends on tolerance factors ,28. Antiolerance ,31. Freqance [Pm . Due to PmS (Pasteurella multocida sensitive to ENR) and PmR (Pasteurella multocida resistant to ENR) were isolated from the lungs of dead cattle that have suffered from BRD in Shuangyang City, Jilin Province. The highly resistant strain (PmHR MIC of ENR is 64 μg/mL) was obtained according to the methods reported in previous studies [PmS, PmR and PmHR had the same molecular type after PFGE typing (previous research of our group) [Pm was cultured on a brain–heart infusion (BHI) medium. Commercial antibiotics were purchased from Sigma-Aldrich Company. According to CLSI regulations, 20,480 μg/mL storage solution was prepared and frozen at −20 °C for later use. Growth was monitored by measuring the OD600 of the liquid cultures using a Biophotometer .The clinical  studies . The PmSr group) . Pm was Pm cells were inoculated into BHI media and incubated for 24h at 37 °C. Then, cells were harvested and washed three times with PBS. Each strain was inoculated at a density of 105 CFU/mL in BHI containing 0.03 to 512 µg/mL of ciprofloxacin and enrofloxacin, and grown for 24 h at 37 °C. The breakpoints used for fluoroquinolone were adopted from the CLSI document, VET01-A4. Escherichia coli ATCC25922 was used as a quality control strain.The minimum inhibitory concentration (MIC) was established by broth microdilution and two-fold agar dilution, as recommended by the Clinical and Laboratory Standard Institute guidelines in VET01-A4. The MICs of ciprofloxacin and enrofloxacin were tested in the range of 0.03 to 512 µg/mL. The gyrA, gyrB, parC and parE genes, were amplified by polymerase chain reaction (PCR) and compared using the gyrA-F (5’-CCTTATCGTAAATCCGCTCGTA-3’), gyrA-R (5’-CGCAGGGACTTTGGTTGG-3’), gyrB-F (5’-GAAATGACCCGCCGTAA-3’), gyrB-R (5’-CTTGCCTTTCTTCACTTTGTA-3’), parC-F (5’-TACGAAGGCATTGAACAAAC-3’), parC-R (5’-CCACTGTCCCTTGCCCTA-3’), parE-F (5’-AATACGGTAAAGCGGTGGC-3’) and parE-R (5’-GGATTCTGCTGGCGGTTC-3’) primers [Fluoroquinolone QRDR targets,  primers . Amplifi600 = 0.5) and treated with ENR (sub-inhibitory concentration) over a period of 15 min. Total RNA extraction of PmS, PmR and PmHR was performed using an RNAiso Pure RNA Isolation Kit  Co., Ltd., China), according to the manufacturer’s instruction. The quality and quantity of RNA was assessed, and the library construction and sequencing were performed at Biomarker Technology  using the Illumina HiSeq 2500 platform. The genome sequence and annotation information of Pm were obtained from the NCBI database (Accession No. NC_014259.1). Quality-filtered reads were aligned to the reference genome sequence, using the CLC Genomics Workbench 4.0 . The cells were grown to an exponential phase  was used for differential expression analysis. The DEGs between the two samples were selected using the following criteria: (i) the logarithmic of fold-change should be greater than 2 and the false discovery rate (FDR) should be less than 0.05. To understand the functions of the differentially expressed gene, GO functional enrichment and KEGG pathway analyses were carried out by Goatools  and KOBAS , respectively. DEGs were significantly enriched in terms of GO and metabolic pathways when their Bonferroni-corrected p-value was less than 0.05. Then, with the increase in drug resistance, differentially expressed genes were screened. To identify DEGs  among the three different samples, the expression level for each transcript was calculated using the fragments of RPKM (read per kilo base per million mapped reads) method. edgeR  and treated with ENR (sub-inhibitory concentration) over a period of 15 min. Total RNA extraction of PmS, PmR and PmHR was performed using an RNAiso Pure RNA Isolation Kit (TaKaRa), according to the manufacturer’s instruction, and was reverse-transcribed to cDNA. The real-time PCR process was performed on the Applied Biosystems 7500 Real-Time PCR System . The primers of satP-F (CGCCAAAAGCAAACGCCATACC) and satP-R (TACGCCAATCCGGGTCCATTAGG) were designed. The specificity of the qRT-PCR results was confirmed by agarose gel and melting curve analysis, and the data were analyzed by the Applied Biosystems™ 7500 software. All qPCR reactions were performed with six replicates.Expression of satP gene knockout was conducted with the suicide vector, pRE112. Briefly, the genome of the strain to be knocked out was amplified by PCR using UP-F/R and DOWN-F/R primers  was cloned into the PRE112 vector, and no base mutation was identified by sequencing. Then, the plasmid pRE112-satP was successively transformed into DH5α-λpir and E. coli WM3064. The single colony was inoculated into 5 mL BHI broth, then inoculated into the BHI agar medium containing 10% sucrose, and cultured overnight at 37 °C. The recombinant strain without the satP gene and pRE112 plasmid was screened . The complemented strain, CPmS, was constructed with the expression plasmids pBBR1-MCS, by homologous recombination.The  primers , respect99) test (the minimum duration for killing 99% of cells), respectively. In short, wild-type strains, deletion strains and complemented strains were cultured to OD600 = 0.5 and colonies were counted. The bacterial solution was diluted to a final concentration of 106 CFU/mL according to the counting results. The final concentration of ENR with MIC was added to the culture medium, 50 μL per hour was taken, and the colonies were counted after dilution. The line diagram of MDK99 was made with the number of colonies as abscissa and time as ordinate. The experiment was set up three times. At the same time, 200 μL per hour was added into a 96-well plate, diluted 10 times continuously, and participated in dilution 5 times. The diluted bacterial solution was inoculated on the BHI solid plate through a multi-point inoculator. The growth of the colonies was observed the next day and the results recorded. Tolerance was determined according to Westfall’s and Cirz’s method ,34. The Female BALB/c mice, aged 6–8 weeks (18–20 g), were obtained from the Animal Experimental Center of Jilin University . All experiments were carried out under the Regulations of Jilin Laboratory Animal Management Committee of Jilin Province. The license number of experimental animals was JLAU20210423001, which is certified by Jilin Science and Technology Association.PmS with bacterial counts of 108, 107, 106, 105 and 104 CFU/mL were intraperitoneally injected in mice. The condition of the mice was observed within seven days and the doses leading to the death of the mice were recorded. The LD50 of the PmS strain was calculated by the Bliss method. The LD50 of the ΔPmS and CPmS strains was determined by the same method as that of the PmS strain. Twenty-five mice, both male and female, were randomly divided into five groups, with five mice in each group. After 3 days of adaptive feeding, the mice were reared without food and water for 12 h, and then Pm was diluted to different concentrations, and each strain was set with five concentration gradients. The infection was carried out by intraperitoneal injection, and each mouse was infected with 100 μL. The control group was injected with the same dose of sterile PBS buffer. After 48 h of observation, the death rate was noted. The LD50 of each strain in the mice was calculated by the Bliss method, using the SPSS 26.0 software.Fifty mice, half male and half female, were randomly divided into five groups, with 10 mice in each group. After adaptive feeding for 3 days, the mice were reared without food and water for 12 h. According to the results of the preliminary experiment, each strain of bovine type A 50. The first and second groups were injected with PmS bacterial solution, the third and fourth groups were injected with ΔPmS bacterial solution, and the fifth and sixth groups were injected with the CPmS bacterial solution.Sixty mice, half male and half female, were randomly divided into six groups, with 10 mice in each group. After 3 days of adaptive feeding, the mice were kept without food and water for 12 h. The bacterial solution was centrifuged to remove the culture medium, then resuspended with normal saline, and the mice were injected with twice the concentration of LDOne hour after bacterial injection, mice in the first, third and fifth groups were intramuscularly injected with a 100 μL of ENR (2.5 mg/kg body weight) twice a day. The second and the fourth groups were injected with the same dose of normal saline intramuscular, and the survival conditions of the mice in each group were observed and recorded, and plotted by the Prism software, according to the previously reported methods ,35.PmS and PmR) with the same molecular typing and different fluoroquinolone resistance phenotypes were isolated by 16S rRNA and PFGE - and verapamil-sensitive efflux. The MICs of ENR were determined using the agar dilution method in the presence and absence of CCCP  and verapamil . These results indicated that the efflux pump genes did not play any role in the quinolone resistance of Pm, we sequenced the transcriptome of three strains under the sub-inhibitory concentration of ENR | > 1, a total of 1,499 significant DEGs were identified. Simply put, compared with  in PmHR . The GO  in PmHR . On the  in PmHR . PmR and PmHR, significantly expressed genes were screened with the improvement of drug resistance, and the satP (gene 1687), focA, napF, glgP and napD genes were significantly up-regulated  with different resistance phenotypes in relation to fluoroquinolones were screened. We found that when the satP gene was deleted, ΔPmS, ΔPmR and ΔPmHR had no QRDR mutation, compared with those without deletion. The different levels of resistance between PmHR and PmR might be mediated by other resistance mechanisms or tolerance. Relevant research has shown that antibiotic tolerance can increase the probability of antibiotic resistance [Pm with different levels of drug resistance, treated by sub-inhibitory concentration, and counted the differential genes. On the basis of ensuring stable and accurate transcriptome data, we analyzed the functional annotation and metabolic pathways, and analyzed and screened the genes that might be related to the formation of drug resistance. The results showed that with the increase in drug resistance, the expression of some genes, namely satP, focA, napF, glgP and napD, increased significantly. The most significant difference was the satP gene, which was up-regulated 17 times in the drug-resistant group and 20 times in the highly drug-resistant group compared with the sensitive group. satP is a representative member of the AceTr family, which is preliminarily identified as a succinate acetate/proton cotransporter transmembrane protein [lamB, and found that it can produce antibiotic resistance by reducing the pressure of protein translation and DNA replication. In order to further detect antibiotic resistance, the lamB gene was knocked out and was found to increase antibiotic resistance by reducing the activity of metabolic pathways such as the tricarboxylic acid (TCA) cycle, pentose phosphate pathway and glycolysis/gluconeogenesis. By comparing the function of the wild-type strain and deletion strain after ciprofloxacin treatment, it was found that the down-regulation of the lamB gene can increase the resistance of bacteria to antibiotics [In this study, three strains (sistance . With re protein . Indraji protein . Yamamot protein . Lu et aibiotics .satP gene, in order to further confirm the function of the satP gene, we constructed the gene deletion strain of a sensitive strain and drug-resistant strain. The results showed that the tolerance of satP decreased after the deletion of the satP gene. Safi et al. have pointed out that when antibiotics act on bacteria, antibiotic tolerance first appears. When antibiotics continue to act, antibiotic tolerance will aggravate the generation of drug resistance. Compared with the life cycle of drug resistance phenotype, antibiotic tolerance is a temporary state. Timely release of antibiotics can restore its tolerance, so the tolerance is reversible [Pm, Westfall et al. studied the mechanism of MRSA tolerance by using triclosan, an antibacterial agent synthesized by fatty acids, to induce the change of MRSA tolerance. MDK99 and the agar diffusion method were used to detect its tolerance. The results showed that triclosan increased the tolerance of LL to bactericidal antibiotics by 10,000 times, and reduced the efficacy of antibiotics by 100 times. Genetic analysis showed that the antibiotic tolerance mediated by triclosan needs the cooperation of ppGpp [satP-gene-mediated tolerance; other membrane transporters have been confirmed to be involved in the formation of tolerance. Poudyal et al. found a MerR-type transcriptional activator, which can activate the BrlR target encoding ABC transport system gene and affect the drug tolerance of bacteria [Based on the research background of the versible . Repeateversible , which rof ppGpp . Howeverbacteria . One of bacteria .satP gene deletion strain was several times lower than that of the wild-type strain, and we found that the treatment rate of the mice infected with ΔPmS was higher. This finding broadly supports the work of other studies in this area which links tolerance with virulence [MazEF gene of three strains of bacteria and found that the tolerance of the deletion strain was decreased and the virulence was enhanced, compared with the wild-type strain [satP can be used as a target of drug resistance inhibitors and how effective it could be in clinical application. More attention should be paid to studying satP and its inhibitors.One unanticipated finding was that the virulence of the irulence . Howevere strain . These fPm, the satP gene was obtained, of which the expression was significantly up-regulated with the increase in drug resistance. Through the study of the formation of the satP gene of drug resistance, the deletion of the satP gene in Pm can lead to a faster growth rate, longer development time of drug resistance and reduced tolerance. In addition, due to the deletion of the satP gene, its virulence was 400 times reduced than in the wild-type strain. The results indicated that satP might be the target of ENR resistance inhibitors. In this study, based on the transcriptome sequence of three strains of


Leiodinychus orbicularis) is a nidicole, i.e., a species that inhabits the nests of various animals. This article also analyses the habitat preferences of the mentioned species and another Uropodina species associated with nests—Apionoseius infirmus. It was proven that L. orbicularis clearly dominated both in the examined dormouse, bat, and bird boxes, whereas A. infirmus, which was less numerous in the communities, preferred natural nests, including the nests of birds of prey. The clear dominance of L. orbicularis in the examined boxes can be explained by the specific microclimate, such as very low humidity, which prevails in the boxes.Hanging nest boxes, which are used by various groups of animals such as birds or mammals , increase the number of shelters and breeding places for these often rare animals. Nest boxes not only become habitats for the host, but are also inhabited by various groups of invertebrates, including insects, spiders, millipedes, snails, and also small arachnids, which are mites. In this article, we present an analysis of the community of one of the groups of mites—Uropodina—which also inhabit nest boxes. In the examined boxes, five species belonging to the discussed group were found, out of which only one (Leiodinychus orbicularis (C. L. Koch) and Apionoseius infirmus (Berlese), i.e., two typically nest-dwelling species of Uropodina. The material for the study was collected from nest boxes in six forest complexes in southwestern Poland. The conducted research revealed the presence of five species of Uropodina, with a total number of 559 specimens, in the examined boxes. Leiodinychus orbicularis was found in almost half of all of the examined boxes and was a superdominant species in the communities. The analysis of the habitat preferences of the two nest species of Uropodina showed that A. infirmus preferred old natural nests, in which the communities were formed from a larger number of species, without a significant statistical prevalence of one species. On the other hand, L. orbicularis occurred sporadically in open bird nests, but was very numerous and frequent in nest boxes. The significant dominance of L. orbicularis in nest boxes can probably be explained by the specific conditions prevailing in this type of microhabitat, including the very low humidity and food resources that this mite species prefers compared to other species of Uropodina.Bird and mammal nests and nest boxes constitute microenvironments in which various groups of invertebrates can live, including mites from the suborder Uropodina (Acari: Mesostigmata). The main aim of the current study was to ascertain the characteristics of mite communities from the suborder Uropodina, which inhabit the nests of dormice (Gliridae) built in nest boxes. The second aim of the study was to compare the habitat preferences of  Every sThe nesting abilities of secondary cavity nesters, bats, and Gliridae mammals depend on the presence of natural cavities that are necessary for establishing nests ,3,4,5. TNest boxes may contain different materials of organic origin. Typically, their most considerable portion consists of nest material, which may be composed of both plant  and animal components , and other remains left by the host, including faeces, pellets, food storages, and remnants after broods (egg shells and dead juveniles) . Such diTalpa europaea L.), marmot (Marmota marmota latirostris Kratochvíl), and badger (Meles meles L.)) [Previous studies have shown that the microhabitats of bird and mammal nests are often also inhabited by mites from the suborder Uropodina (Acari: Mesostigmata). Uropodina mites were found both in the nests and nest boxes of various bird species ,41,42,43les L.)) ,46. Thesles L.)) , which eles L.)) . The facles L.)) .Phylloscopus sibilatrix (Bechstein)), the passerine species, which builds its nests on the ground, have revealed that there is a lack of typical nidicoles in such nests and that the community structure is very similar to those found in the soil and nests of the common mole [It is also worth mentioning that most of the research conducted so far has focused mainly on Uropodina communities inhabiting arboreal or aboveground bird nests ,53,56,57mon mole ,46 compaLeiodinychus orbicularis [Phaulodiaspis rackei (Oudemans), Ph. advena (Trägårdh), and Ph. borealis (Sellnick), which inhabit underground nests of the mole, the marmot, and the European water vole (Arvicola amphibius L.) [Apionoseius infirmus (Berlese) is often and numerously found, while Nenteria pandioni Wiśniewski et Hirschmann occurs exclusively in the nests of the white-tailed eagle . Nest boxes inhabited by dormouse mammals have not yet been studied for the presence of Uropodina mites.The Uropodina species, which is most frequently and abundantly found in the nests of various bird and mammal species, is icularis , describbius L.) . In nestGlis glis L.), the garden dormouse (Eliomys quercinus L.), the forest dormouse ), and the hazel dormouse (Muscardinus avellanarius L.), all of which are legally protected, with some still requiring active protection. The presence of dormice is associated with the presence of deciduous and mixed forests of high natural value, with an availability of trees with hollows, in which they shelter, reproduce, and store food [Dormice (Gliridae) are a family of mammals from the suborder Sciuromorpha in the order Rodentia. In Poland, they are represented by four species, the European edible dormouse  built in nest boxes. In addition, an analysis of the occurrence and ratio of two nest species of Uropodina, i.e., This study was conducted in six forest complexes in the central and southern parts of Opolskie voivodeship (south and southwestern Poland). Two sites are located in the Opawskie Mountains, and the four other sites are located in the lowland part of the Opole region, that is, in the Stobrawski Landscape Park and Niemodlin Forest , and the identification of the extracted species was conducted by the first author on the basis of the publications by Karg , BłoszykL. orbicularis and A. infirmus in nests of different hosts  and the frequency of occurrence (F). The scale has the following classes: dominance D5 eudominants (>30.0%), D4 dominants (15.1–30.0%), D3 subdominants (7.1–15.0%), D2 recedents (3.0–7.0%), and D1 subrecedents (<3.0%); frequency F5 euconstants (>50.0%), F4 constants (30.1–50.0%), F3 subconstants (15.1–30.0%), F2 accessory species (5.0–15.0%), and F1 accidents (<5.0%) . The avent hosts  were stop < 0.05. All probability values shown here are two-tailed. All statistical analyses followed the formulae in STATISTICA 12.0 [We used non-parametric tests (Fisher’s exact test and Mann–Whitney U test). The established significance level in the statistical analysis was ICA 12.0 .Leiodinychus orbicularis turned out to be the most numerous species. A total of 559 specimens of this species were found, including 184 females, 153 males, 165 deutonymphs, and 57 protonymphs. Moreover, L. orbicularis was the superdominant species in the examined community and was present in nearly half of the examined nest boxes. It was present both in bird nest boxes (11 boxes) and dormice nest boxes (6); there were no differences in the type of box selection . The mean number of L. orbicularis also did not differ between the types of nest boxes . In individual boxes, the presence of 1 to 285 specimens was recorded   . L. orbicularis occurs in most of the merocenoses of this type that have been analysed so far . This means that the specific microclimate in the boxes is the factor that attracts this mite species.In the examined boxes inhabited by dormice, the occurrence of five species of Uropodina was recorded, of which only one, i.e.,  species ,34,55. Tronments . Occasiost boxes ,34. The L. orbicularis monopolises all available resources . However, these are usually communities with a low number of species, in which esources . In typiesources . Haliaeetus albicilla (L.))), ground nests of the wood warbler, and underground burrows of the badger, and the low percentage of the specimens in other underground mammal nests is probably due to the method of dispersion of the species, namely the deutonymphs of L. orbicularis (with pedicels), which are found in nests and are carried by insects [L. orbicularis determine the preferred type of the nests inhabited by this species, excluding those located underground. It is worth mentioning that the discussed species is characterised by a wide ecological valence, which allows it to occupy various niches, including those created by humans, and for this reason, it was also found in stored products [The absence of this species in the nests of birds of prey ), and red kite (Milvus milvus (L.)), the abundance of this species was higher than that of L. orbicularis. Generally, it can be stated that unlike L. orbicularis, A. infirmus avoids nests built in nest boxes, but it is more often present in old natural nests, where there are usually more species in the community, without a clear statistical prevalence of one species.The second of the analysed nidicoles, i.e., starling . BesidesL. orbicularis in the examined boxes (regardless of the host that inhabited them) can be explained by the specific microclimate that prevails in the boxes [Apparently, little is known about the method of dispersion of nidicoles and the routes by which they reach isolated microhabitats, such as nest boxes. The presence of phoretic deutonymphs suggests that the discussed species of Uropodina are carried by insects, probably beetles (like most phoretic Uropodina). However, specific vector species have not been found yet. The clear dominance of he boxes . This sp


Inflammation and thrombosis are involved in the development of stroke-associated pneumonia (SAP). Our aim was to evaluate the predictive value of a novel, simplified, thrombo-inflammatory prognostic score (TIPS) that combines both inflammatory and thrombus biomarkers in the early phase of ischemic stroke (IS).The study population consisted of 897 patients with a first diagnosis of IS admitted to the emergency department of five tertiary hospitals in China. Of these, the data from 70% of patients was randomly selected to derive the model and the other 30% for model validation. A TIPS of “2” was indicative of high inflammation and thrombosis biomarkers and “1” of one biomarker, with “0” indicative of absence of biomarkers. Multivariate logistic regression analyses were used to identify the association between TIPS and SAP.2DS2) and biomarkers currently used in practice, for both the derivation and validation sets. Mediation analysis revealed that TIPS provided a predictive value than either thrombotic (NLR) and inflammatory (D-dimer) biomarkers alone.The TIPS was an independent predictor of SAP and 90-day mortality, with the incidence of SAP being significantly higher for patients with a high TIPS. The TIPS provided superior predictive value for SAP than clinical scores (AThe TIPS score may be a useful tool for early identification of patients at high-risk for SAP after IS. Ischemic stroke (IS), which accounts for more than 2/3 of all strokes, is a life-threatening emergency associated with high-risk for morbidity and mortality, worldwide . Among i2DS2 Score, Kwon Pneumonia Score, PASS Pneumonia Rule, and ISAN Score . The methods of the REMISE study were conducted following the tenets of the Declaration of Helsinki and were approved by the institutional review boards of Sichuan University West China Hospital and other participating hospitals.Our REMISE study was registered at We designed and validated a novel TIPS system to identify patients at high-risk for SAP after an IS early in the stroke center of emergency department (ED). Our study population included patients with acute IS admitted to the stroke centers of five tertiary hospitals in China, from January 2020 to December 2020. Eligible patients were those with a first diagnosis of IS, according to the 2019 American Heart Association Stroke guideline . The incDemographic and clinical patient data during hospitalization were retrieved from the REMISE study database by trained physicians, using standard case report forms. The following data were extracted: age, sex, vital signs, laboratory test results, body mass index, medical history, arterial blood gas analysis, imaging results, adverse outcomes, and therapies received in the hospital. All laboratory tests and imaging examinations were performed using the standard procedures of Sichuan University West China Hospital.2DS2 score is a validated screening tool for SAP, which includes the predictive factors of age, dysphagia, male sex, atrial fibrillation, and stroke severity, with a higher score being predictive of a higher risk for SAP  , with hi for SAP . The pne for SAP .The primary outcome was SAP, diagnosed according to the Centers for Disease Control and Prevention standards of the Outcome Evaluation Committee. The occurrence of SAP was identified based on the electronic medical records of the REMISE study.The TIPS used in our study was calculated based on inflammatory  and thrombotic  biomarkers measured at the time of admission. The presence of high inflammation and thrombosis biomarkers was indicative of high-risk for SAP, quantified by a TIPS score of 2. The presence of only one or absence of inflammation or thrombosis biomarkers was assigned a TIPS of “1” and “0,” respectively.2) test or Fisher’s exact test was used for between-group comparison of categorical data.Of the patients included, the data from 70% were randomly used to derive the model (derivation set), with the other 30% used to validate the model . Classification variables were calculated as a frequency and percentage, with continuous variables calculated as the mean ± standard deviation or median and interquartile range (IQR). An analysis of variance was used to compare parametric patient characteristics and the Kruskal–Wallis H test to compare non-parametric patient characteristics between the SAP and non-SAP groups. The chi-squared  and R Studio .Cut-off values for the different inflammatory  and thrombotic  biomarkers, used to calculate the TIPS, were evaluated using receiver operating characteristic (ROC) analysis and determined by the maximal corresponding values of Youden’s index. To evaluate the predictive value of TIPS for SAP, the area under the ROC curve (AUC), integrated discrimination improvement (IDI), and net reclassification index (NRI) were calculated for both the derivation and validation sets to identify discrimination . Spearmaerformed . For alln = 246) and derivation (n = 651) sets and the mean A2DS2 score was 4 (IQR: 2–6) for both the derivation and validation sets. A total of 238 (36.5%) and 81 (32.9%) patients had SAP after IS in the derivation and validation sets, respectively.In total, 897 patients with IS met our criteria for inclusion, with a mean age of 66 ± 13 years and 63.1% of the study sample being men. There were no differences in the demographic and clinical characteristics between patients in the model validation  or thrombotic biomarkers  in both the derivation and validation sets . Thus, NThe demographic and clinical characteristics of the derivation set according to the TIPS are shown in p = 0.016 and TIPS 2 vs. 0: OR: 7.494, 95%CI: 4.402–12.757, p < 0.001) and the validation set   .2DS2  in the derivation set, with similar results for the validation set (The AUC for the prediction of SAP using TIPS was 0.762 (95%CI: 0.725–0.802), which was larger than for the Ation set .2DS2 score was calculated to confirm the improved discrimination of TIPS  and the validation set  than NLR  and D-dimer  in the derivation set, with similar results were observed in the validation set  and SAP, with the mediating effect of thrombo-inflammatory on SAP being higher than for a single inflammatory or thrombotic state. Consequently, our results support the TIPS as a useful tool for the early identification of high-risk SAP in patients with IS at admission  (p < 0.001)  . The NLR 0.0001) . As impo 0.0001) . As an e< 0.001) . The res< 0.001) . In our Fibrinogen, platelet count, INR, APTT, prothrombin time, and D-dimer in SAP patients were significantly higher than those in non-SAP patients. The fibrinogen-to-albumin ratio  has been reported to be an independent predictor of high-risk of SAP , with both the FAR and platelet-to-lymphocyte ratio (PLR) being associated with the severity of SAP . Therefo2DS2) and single biomarkers of thrombo-inflammation, irrespective of the severity of IS.Inflammation surges immediately after an injury, leading, in the acute phase, to excessive activation of the coagulation system, with the resulting coagulation state greatly increasing the inflammatory process . TherefoThe limitations of our study need to be acknowledged. First, this was a single-center retrospective cohort study and, thus, causal effects between the TIPS, its associated thrombo-inflammatory biomarkers, and SAP cannot be inferred. Second, model derivation and validation was based on biomarkers measured in the ED at the time of admission, with changes in these biomarkers, measured at different time points, not considered. As such, the value of TIPS to predict SAP at different time points post-IS was not evaluated. Third, we did not evaluate the predictive value of TIPS in IS patients in some subgroups including different kinds of therapies, and more than 6 h of symptom onset to hospitalization, and so on. Forth, some additional inflammatory thrombus biomarkers were not used in the TIPS model derivation. Lastly, we did not evaluate the time required to complete the TIPS during admission in the ED. Therefore, further multicenter, prospective, studies are needed to confirm the predictive value of TIPS for SAP and its application in practice.2DS2 score. Therefore, the TIPS score is a useful scoring system to identify patients who are at high-risk for SAP after an IS and, as a supplementary tool, it could improve the predictive power of traditional SAP risk scores. Further well-designed studies are required to confirm these results.In summary, we provide findings of the predictive value of the TIPS, which is based on thrombo-inflammatory biomarkers for SAP in the early phase of IS onset. We show that an elevated TIPS was an independent predictor of SAP after an IS. In addition, the discrimination and accuracy of SAP were superior to those of the Azengzhi2013@qq.com.The data analyzed in this study is subject to the following licenses/restrictions: The data that support the findings of this study are available on request from the corresponding author. Requests to access these datasets should be directed to ZZ, The studies involving human participants were reviewed and approved by the institutional review boards of Sichuan University West China Hospital. Written informed consent to participate in this study was provided by the participants’ legal guardian/next of kin.DL, ZZ, and YC conceived the study design. DL, YJ, JY, YL, XC, HL, and ZW collected the epidemiological and clinical data. DL, YJ, YL, and ZW summarized the data and performed the statistical analyses. DL, YL, and YC interpreted the data and drafted the manuscript. LY and ZZ participated in the design of the study, acquired the data, and helped to revise the manuscript. All authors have accepted responsibility for the entire content of the submitted manuscript and approved its submission.This work was supported the Sichuan Science and Technology Program (Nos. 2022YFS0279 and 2021YFQ0062), Sichuan Provincial Health Commission (No. ZH2022-101), and Sichuan University West China Nursing Discipline Development Special Fund Project (No. HXHL21016).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


The objective of this cross-sectional study was to investigate health management, well-being, and pandemic-related perspectives of chronic disease patients in the context of stringent measures, and associated correlates. A self-report survey was administered during the Omicron wave lockdown in Shanghai, China. Items from the Somatic Symptom Scale (SSS) and Symptom Checklist-90 were administered, as well as pandemic-related items. Overall, 1,775 patients  were recruited through a community family physician group. Mean SSS scores were 36.1 ± 10.5/80, with 41.5% scoring in the elevated range . In an adjusted model, being female, diagnosis of coronary artery disease and arrhythmia, perceived impact of pandemic on life, health condition, change to exercise routine, tolerance of control measures, as well as perception of future and control measures were significantly associated with greater distress. One-quarter perceived the pandemic had a permanent impact on their life, and 44.1% perceived at least a minor impact. One-third discontinued exercise due to the pandemic. While 47.6% stocked up on their medications before the lockdown, their supply was only enough for two weeks; 17.5% of participants discontinued use. Chief among their fears were inability to access healthcare (83.2%), and what they stated they most needed to manage their condition was medication access (65.6%). Since 2020 when we assessed a similar cohort, distress and perceived impact of the pandemic have worsened. Greater access to cardiac rehabilitation in China could address these issues. While tMeasures to control infection spread, such as physical distancing often necessitating closure of essential businesses , as wellGovernment policy varies worldwide in terms of implementation of infection prevention and control measures. Based on the COVID-19 stringency index, such strategies are among the most stringent in China . For exaGiven these unprecedented circumstances, assessment of the impacts of these strict control measures on chronic disease patients is needed, particularly in the context of some of the most stringent control measures globally, in one of the most densely populated cities in the world where risk of transmission is hence greater ,14, and 22.1Study approval was secured from the Ethics Committee of Xinhua Hospital, affiliated with Shanghai Jiaotong University School of Medicine (XHEC-C-2022-042). The anonymous online survey in Simplified Chinese was created by the Cardiac Rehabilitation (CR) group, Health Risk Assessment and Control of the Chinese Preventive Medical Association. Data collection for this cross-sectional study was undertaken between March and June 2022. It was distributed via Wenjuanxing by family doctors who belonged to the Community Physician group of the Psychosomatic Medicine Special Committee of the Shanghai Association of Integrated Chinese and Western Medicine; this group cooperates closely with the Chinese Preventive Medical Association.2.2During the Omicron wave lockdown in Shanghai, residents including patients with chronic conditions were to remain indoors and could not go outside to exercise or purchase groceries at the supermarket. They could receive care via telehealth or consultation in internet hospitals. Medication was available through online pharmacies with in-home delivery. This could be challenging for seniors who may have low digital literacy or who do not have access to a smartphone.To receive in-person care, health codes had to be scanned. Daily nucleic acid tests were mandatory for in-patients and the one allowed family member who could accompany a patient.Participant inclusion criteria were the following: outpatients with cardiovascular and cerebrovascular diseases, diabetes, kidney, or another chronic condition . Participant exclusion criteria were: (1) outpatients with severe cognitive impairments or any conditions that prevent them from being able to understand and agree to participate in the research; (2) outpatients unwilling to cooperate with the research protocol; and (3) outpatients with any missing data within questionnaire items.2.3All items were self-reported. Two relevant and validated psychosocial scales available for use in China were selected by the CR group, assessing depressive and anxiety symptoms, somatic symptoms, as well as sleep and cognitive issues. First, five items from the validated Symptom Checklist-90 were administered to assess depressive symptoms ,16. TherThe Chinese version of the Somatic Symptom Scale (SSS) was also administered . It compIn addition, non-psychometrically validated items related to pandemic perceptions and health management were generated by the CR group, based on their knowledge and expertise. Response options were forced choice, with some where respondents could select all that apply. Participants were asked to respond these items based on their experience in the prior 2 weeks.2.4p < 0.05 considered statistically significant. Descriptive analysis was first performed. Perceptions of impact of the pandemic (at least minor vs none) on their chronic condition and psychosocial well-being were tested using chi-square and t-tests as applicable. The association of pandemic-related perceptions with SSS scores was assessed using the t-test and chi-square, as applicable. Finally, associations between participant sociodemographic and clinical characteristics with SSS scores were tested using logistic regression analysis as applicable. Then, an adjusted model was computed for the association of the SSS with those sociodemographic, clinical, and pandemic-related perceptions that were significant at p < 0.05.IBM SPSS statistics version 25.0 was used for all statistical analyses, with 3Of 1,775 responding patients, most were educated, working, married females living with family, and who had healthcare insurance . As showp ≤ 0.016). All chronic conditions except hypertension and diabetes were associated with significantly more distress on the SSS , and they were living longer with their condition (p < 0.001). They were significantly more likely to have suffered a major life event .The mean psychosocial well-being scores are shown in 3.1As shown in Half perceived the pandemic had at least a minor impact on their health or more . As showp = 0.009) and who had lower somatic (p < 0.001) were more likely to change their exercise due to the pandemic and associated control measures (no association with chronic condition).Forty percent exercised at their home, but over one-third stopped exercising and one-fifth changed their exercise . ParticiWith regard to medication, while almost half tried to stock up before the lockdown, almost one-fifth of participants discontinued use . Only juParticipant’s greatest worries related to Omicron and their chronic condition are also shown in Pandemic perceptions were highly associated with SSS . Partici4This is the first or one of the first full studies to examine the impact of the COVID-19 pandemic and its associated control measures in a large sample of older adults with chronic conditions . It was The burden of distress was quite high, even when considering that rates of psychological distress are higher in those with chronic disease ,22 and rIn terms of health management, despite the proven benefits of secondary prevention behaviours, such as exercise and medication adherence , many chIn our earlier examination of the impact of the pandemic and associated control measures on older adults  in the spring of 2020 , mean scStudy implications relate to ensuring that patients have what they need to manage their condition. What they most wanted was easier access to healthcare and medications, regardless of control measures. This is despite quite fast and broad availability of telehealth in China and online pharmacies . This coCaution is warranted when interpreting these results. First, this is a cross-sectional study, so the design precludes causal determinations. Second, the results may not be generalizable beyond China with its political and cultural context, and the city of Shanghai more specifically where the stringency of control measures was high. Nevertheless, the population of Shanghai is very large, and findings can inform other governments when considering their control measures. Moreover, similarity of the sample to the larger population is not known given the recruitment strategy, designed to quickly secure data during the lockdown, but where full reach and number of non-respondents were not collated. Third, all data were self-report, and hence, there may have been socially desirable responding or other measurement error. Relatedly, psychiatric conditions and health status were not verified through structured clinical interview or medical records. Fourth, due to the exploratory nature of the study, multiple comparisons were performed, increasing the chance of type 1 error. Finally, whether patients had COVID-19 prior to or during the period of study was not considered; more research is needed.In conclusion, 2 years into the pandemic in the midst of arguably the most stringent prevention and control measures globally, over 40% of chronic disease patients are experiencing elevated psychosocial distress, and this was associated with female, diagnosis of coronary artery disease and arrhythmia, perceived impact of pandemic on life, and impact of pandemic on health condition . Many pa


Early judgment of the progress of acute pancreatitis (AP) and timely intervention are crucial to the prognosis of patients. The purpose of this study was to investigate the application value of CT-based radiomics of pancreatic parenchyma in predicting the prognosis of early AP.n = 95) and a validation set (n = 42) in a ratio of 7: 3. The region of interest (ROI) was outlined along the inner edge of the pancreatic parenchyma manually, and the Modified CT Severity Index (MCTSI) was assessed. After resampling and normalizing the CT image, a total of 2,264 radiomics features were extracted from the ROI. The radiomics features were downscaled and filtered using minimum redundancy maximum correlation (mRMR) and the least absolute shrinkage and selection operator algorithm (LASSO) regression, in turn, and the more optimal subset of radiomics features was selected. In addition, the radiomics score (rad-score) was calculated for each patient by the LASSO method. Clinical data were also analyzed to predict the prognosis of AP. Three prediction models, including clinical model, radiomics model, and combined clinical–radiomics model, are constructed. The effectiveness of each model was evaluated using receiver operating characteristic (ROC) curve analysis. The DeLong test was employed to compare the differences between the ROC curves. The decision curve analysis (DCA) is used to assess the net benefit of the model.This retrospective study enrolled 137 patients diagnosed with AP (95 cases in the progressive group and 42 cases in the non-progressive group) who underwent CT scans. Patients were randomly divided into a training set  and the radiomics model  in the validation set. Decision curve analysis demonstrated higher net clinical benefit for the clinical–radiomics model.The mRMR algorithm and LASSO regression were used to select 13 radiomics features with high values. The rad-score of each texture feature was calculated to fuse MCTSI to establish the radiomics model, and both the clinical model and clinical–radiomics model were established. The clinical–radiomics model showed the best performance, the AUC and 95% confidence interval, accuracy, sensitivity, and specificity of the clinical–radiomics model in the training set were 0.984 (0.964–1.000), 0.947, 0.955, and 0.931, respectively. In the validation set, they were 0.942 (0.870–1.000), 0.929, 0.966, and 0.846, respectively. The Delong test showed that the predictive efficacy of the clinical–radiomics model was higher than that of the clinical model (The pancreatic parenchymal CT clinical–radiomics model has high diagnostic efficacy in predicting the progression of early AP patients, which is significantly better than the clinical or radiomics model. The combined model can help identify and determine the progression trend of patients with AP and improve the prognosis and survival of patients as early as possible. Acute pancreatitis (AP) is an inflammatory disease of the exocrine pancreas with a complex and variable clinical course. It is a common acute abdomen of the digestive system. The pathogenesis is due to the abnormal activation of pancreatic enzymes that destroy the pancreas itself and surrounding tissues and organs, with the local inflammatory infiltration of the pancreas as the main feature. The etiology of AP is diverse, mainly biliary tract diseases, alcoholism, hypertriglyceridemia, and less commonly drugs, endoscopic retrograde cholangiopancreatography, postoperative period, metabolic factors, infections, heredity, autoimmune disorders, and trauma . AccordiPatients can feel severe abdominal pain and often trigger systemic inflammatory response syndrome (SIRS) and multiple organ dysfunction syndrome (MODS) and other severe complications. Then, this triggers pancreatic necrosis and persistent organ failure, which in severe cases can even lead to death with a mortality rate of 1–5% . NumerouTherefore, early prediction of the development of acute pancreatitis and taking reasonable measures timely are essential for the prognosis of patients. The occurrence of complications can be used as a valid indicator for prognostic assessment of pancreatitis . ComplicClinicians diagnose AP mainly by observing patients’ symptoms, signs, and changes in laboratory indicators. However, early clinical symptoms and signs of AP are not specific. Serum amylase and lipase levels do not fully reflect the severity and progression of AP , 9. CRP This study is based on pancreatic CT plain scan images for radiomics analysis and fusion of clinical data. Exploring the prediction of complications in pancreatitis based on clinical–radiomics models to determine the prognosis of early AP.A total of 137 patients from April 2021 to November 2022 at the First Affiliated Hospital of Harbin Medical University were retrospectively collected, and imaging data and relevant clinical laboratory data of patients with a clinical diagnosis of AP were enrolled. Ethics committee approval was granted for this retrospective study, and the requirement for written informed consent was waived. The diagnostic criteria of AP were based on the 2012 revised Atlanta Classification of AP . Moreover, the radiomics feature extraction, feature selection, and machine learning models building were established on the uAI Research Portal , which was integrated with PyRadiomics (version 2.2.0), Scikit-Learn (version 1.2.0), and so on. The CT images were analyzed by two radiologists with 10 and 20 years of clinical experience blinded to outline the region of interest (ROI) along the edge of the pancreatic parenchyma by manual segmentation , and MCTTo ensure repeatability, gray intensity normalization and resampling were performed to eliminate the heterogeneity between different scanners before feature extraction. Images were resampled to 1 mm × 1 mm × 1 mm voxels using the B-Spline interpolation method and normalized by subtracting the window level (WL: 30) and dividing by the window width (WW: 300). The extractable feature groups include first-order features, shape, and texture features . The filter transforms include 14 filters, such as the Laplace-Gaussian filter, wavelet analysis, and local binary mode transform. A total of 2,264 radiomic features were finally extracted from the pancreatic parenchyma.Intraclass correction coefficients (ICCs) were used to calculate intra-observer and inter-observer reproducibility . Two radTo avoid overfitting, it is necessary to reduce the dimensionality of the image data before the establishment of the radiomics label. The obtained radiomic features were reduced and filtered by using mRMR algorithm and LASSO regression to obtain the optimal subset of features, and the rad-score of each patient was calculated for each patient based on feature weights.The obtained rad-score was fused with the MCTSI score to create a radiomics model. The baseline data were used to build a clinical model. A joint clinical–radiomics model was established by combining the two. The AUC values of the three models were analyzed using ROC curves. Compare the performance of different models in the training and validation sets. The performance of the same model is also compared in the training and validation sets. The DeLong test was used to compare the statistical differences among the three models. Evaluate clinical–radiomics characteristics by univariate and multivariate logistic regression analyses. The net clinical benefit of the models was compared by DCA.t-test for comparison of differences between groups; continuous variables that did not obey normal distribution are presented as median , and Wilcoxon rank sum test was used for comparison of differences between groups. Categorical variables were presented as frequency, and differences between groups were compared using the chi-square test or Fisher’s exact test for comparison of differences between groups. p < 0.05 was considered statistically different. The AUC and 95% confidence interval, accuracy, sensitivity, and specificity were used to evaluate the performance of the models, and the DeLong test was used to compare the differences between the ROC curves of the three models. DCA is used to compare net clinical benefit.Statistical analysis was performed using SPSS  and R statistical software (Version 4.1.0). Clinical data for the training set and validation set were analyzed according to the variable type. Continuous variables that obeyed normal distribution are presented as mean ± standard deviation and analyzed by n = 95) and non-progressive (n = 42) groups. There were 95 cases in the training set (66 cases in the progressive group and 29 cases in the non-progressive group) and 42 cases in the validation set (29 cases in the progressive group and 13 cases in the non-progressive group).A total of 137 AP patients were enrolled in this study, which consisted of 84 male and 53 female cases with a mean age of 43  years. Patients were divided into progressive  and PCT  in the progression group were higher than in the non-progression group, while the gender differences were not statistically significant . In the training set, the age in the progressive group was younger than the non-progressive group , and the CRP was higher than the non-progressive group   .Z = –5.3; –3.34, P < 0.001).In both the training set and validation set, the MCTSI scores in the progression group were higher than in the non-progression group  and the radiomics model  , radiomics models , and clinical–radiomics models . The results of DCA indicate that the clinical–radiomics model has a higher clinical net benefit  . There w benefit .CI: 2.259–27.047; p = 0.003), PCT , and rad-score  were independent predictors of progression of acute pancreatitis, and two factors, MCTSI  and CRP , were not independent factors.The results of univariate and multivariate logistic regression analysis of clinical–radiomics features predicting the progression of acute pancreatitis are shown in The initial diagnosis of AP patients accounts for approximately 0.3% of the total number of emergency department patients. The pathogenesis is due to the abnormal activation of pancreatic enzymes. The pancreatic enzyme causes damage to the pancreas itself and surrounding organs, which, in turn, triggers related inflammatory and immune responses. In severe cases, it can lead to organ dysfunction and even death . There ip < 0.05). With the progressing pancreatitis, PCT and CRP also increased. BISAP can early identify patients with a high risk of complications and death using five simple indicators  The analysis of this study was based on radiomics features extracted from two-dimensional images of the pancreatic parenchyma at the largest level of the diseased pancreas, and it may be more effective to reflect the lesions if three-dimensional images of the entire pancreatic parenchyma are extracted; (2) a retrospective single-center study was conducted with hospitalized patients. There may be significant bias in sample selection; (3) AP patients in our hospital undergo routine CT plain scans upon admission. If conducting radiomics analysis based on CECT scanning images, a more efficient model can be obtained; (4) patients with mild symptoms do not need image examinations to diagnose and manage AP. Currently, there are few imaging studies on patients with mild symptoms. Moreover, our results tend to analyze more severe and complicated pancreatitis.In summary, the clinical–radiomics model based on pancreatic parenchymal CT has good predictive performance in both the training and validation sets, helping to identify and judge the progression trend of acute pancreatitis patients as soon as possible, and taking timely and effective measures to improve the patient’s prognosis.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving humans were approved by the Ethics Committee of First Affiliated Hospital of Harbin Medical University. The studies were conducted in accordance with the local legislation and institutional requirements. The ethics committee/institutional review board waived the requirement of written informed consent for participation from the participants or the participants’ legal guardians/next of kin because (1) this study only retrospectively analyzed the patient’s imaging and clinical data, without any intervention on the patient’s condition, and there is no risk to the life and health of the subjects. (2) For the collection or research of previously archived data, files, records, and imaging data, researchers are unable to contact the subjects.MX: Conceptualization, Methodology, Writing – original draft. SL: Data curation, Methodology, Writing – original draft. DX: Data curation, Writing – original draft. HW: Formal analysis, Resources, Writing – original draft. QG: Investigation, Writing – original draft. LZ: Investigation, Writing – original draft. XX: Conceptualization, Project administration, Writing – review & editing. YJ: Methodology, Writing – review & editing.


Previous studies have shown that malnutrition before hematopoietic stem cell transplantation (HSCT) is associated with poor patient prognoses. There is inconsistency among studies on which nutritional status screening tool is appropriate for malnutrition diagnosis before allo-HSCT. The present study aimed to compare nutritional screening tools in patients with leukemia before allo-HSCT.An observational, cross-sectional, and single-center study was conducted in Tehran, Iran. One hundred four adults allo-HSCT candidates aged 18-55 years with leukemia were selected sequentially. Malnutrition assessment was done using three tools, the Global Leadership Initiative on Malnutrition (GLIM), nutritional risk screening 2002 (NRS-2002) and European Society for Clinical Nutrition and Metabolism (ESPEN) criteria. The agreement between malnutrition assessment tools was evaluated with Cohen’s kappa.p < 0.001), while the agreement between GLIM and ESPEN was fair . The agreement between NRS-2002 and ESPEN was fair . We also found a moderate agreement for all tools .The agreement between GLIM and NRS-2002 was perfect (κ = 0.817, NRS-2002 is an accepted tool for screening malnutrition in hospitalized patients. In the current study, the GLIM criterion perfectly agreed with the NRS-2002. Further studies in the HSCT setting are needed to introduce a valid tool. Allogeneic hematopoietic stem cell transplantation  is a usual treatment for hematological diseases like leukemia . Even thFew studies demonstrated nutritional screening tools to diagnose malnutrition in patients with hematological malignancies prior to HSCT, and none has presented conclusive evidence that verified which tool is obviously superior , 11. AmoNotably, the Global Leadership Initiative on Malnutrition (GLIM) criteria were suggested by nutritional scientific societies in 2019 for diagnosing malnutrition . It consUntil now, no study evaluates the malnutrition status of patients with leukemia who are candidates for allo-HSCT with GLIM criteria. A few studies compare the GLIM criteria with other previously utilized criteria, including NRS-2002 and ESPEN. We explained that NRS-2002 offers a holistic assessment, ESPEN’s evidence-based nature ensures a comprehensive framework, and GLIM criteria incorporate the latest advancements. This comparison enables us to identify the most appropriate criteria in the clinical setting. Therefore, this study aimed to compare the diagnostic test accuracy of the GLIM, NRS-2002, and ESPEN criteria for malnutrition diagnosis before allo-HSCT in patients with leukemia. This combination of tools ensures a thorough evaluation of nutritional risk in the context of allo-HSCT.An observational, cross-sectional, and single-center study was conducted in the Hematology Center of Shariati Hospital, Tehran, Iran. One hundred four adult allo-HSCT candidates aged 18–55 years with leukemia hospitalized in bone marrow transplant wards from November 2021 to December 2022 were selected sequentially. Patients with leukemia were eligible for inclusion if they were aged between 18 and 55 and candidates for allo-HSCT.The exclusion criteria were refusal to be involved in this study, and patients who were candidates for autologous transplantation.Demographic variables such as age, sex , and clinical variables of the patients, which included the type of malignancy (acute myeloid leukemia (AML) or acute lymphoblastic leukemia (ALL)), complete remission (CR) status , and risk status  were extracted from the patient’s medical records.The functional status of patients was assessed based on the Karnofsky functional status index, which includes a score from 0 (dead) to 100  with 10-point intervals . A score2).Body weight was measured using a digital scale  with an accuracy of 0.1 kg. Height was assessed using a tape measure attached to the wall and an accuracy of 0.5 cm. BMI was calculated by dividing weight (kg) by the square of height (mThe Tanita (BC-418’s) device was used to evaluate body composition during fasting conditions, with minimal water consumption and little exercise before the test and after defecation. This device offers independent analysis for various organs (right and left hand and right and left foot) and the trunk due to employing eight electrodes (two beneath the right foot and two in the right and left hand). Before the assessment, all the anthropometric devices were calibrated.Daily calorie intake and consumption of macronutrients  of all patients was checked by a trained dietitian with 3-day 24-h dietary recalls . The recorded amount of each food using the scale guide for Iranian households was converted to grams , and ana2 for <70 years and < 22 kg/m2 for ≥70 years) and reduced muscle mass. In the current study, the reduction of muscle mass was considered based on Appendicular Skeletal Muscle Index  <7 for men and ASMI <5.7 for women. Etiological criteria include reduced energy expenditure, chronic gastrointestinal status, disease burden, and inflammatory conditions. According to the definition of GLIM criteria, disease burden, and inflammation were considered positive in all patients in the current study.Nutritional assessment was conducted by a trained evaluator who remained blinded during the initial 48 h of hospitalization. Three tools were utilized for this purpose: the NRS-2002 for nutritional screening to identify patients at risk, and the GLIM and ESPEN criteria for diagnosing malnutrition. The GLIM criteria requires at least one phenotypic and etiological criteria for malnutrition diagnosis . PhenotyAssessment of nutritional status based on NRS-2002 is done using a questionnaire. The main components of this questionnaire were: 1) the severity of the impact of the primary disease on the nutritional status; 2) recent changes in body weight (during the last one to 3 months); 3) Change in food intake during the last week; 4) BMI; If the evaluated person is 70 years old or more, one point will be added to the sum of her points. NRS score equal to 3 and more than 3 is defined as a nutritional risk .2 for patients ≥70 years old and < 20 kg/m2 for <70 years) or low fat-free mass index (FFMI) (<15 kg/m2 in women and < 17 kg/m2 in men)  along with low BMI (<22 kg/m in men) .ncss.com/software/pass. Therefore, taking into account 10% drop-out rate, the sample size was finally determined to be 104 patients .The Kolmogorov–Smirnov test was used to evaluate the normality of continuous variables. Demographic, clinical, and anthropometric variables, as well as dietary intakes of patients in accordance with malnutrition status , were evaluated by independent sample t-test or Mann–Whitney test and Chi-squared or Fisher’s exact test for continuous and categorical variables, respectively. The agreement between the two malnutrition assessment tools was evaluated with Cohen’s kappa . Fleiss’The ethics committee of Tehran University of Medical Sciences evaluated and approved this project (Ethics code: IR.TUMS.MEDICINE.REC.1400.1089). A signed consent form was obtained from all the participants.n = 67, 64.4%). The GLIM and NRS-2002 indicated the highest percentage of patients with malnutrition (26 and 33.6%), and the ESPEN criteria indicated the lowest percentage of patients with malnutrition (8.65%). Patients without malnutrition were more prone to be in the first CR for all tools . The NRS-2002 showed that patients without malnutrition were more prone to AML than ALL (p = 0.049); however, the adverse risk of disease was higher (p = 0.012).p < 0.05). Although GLIM and ESPEN criteria indicated that FM was significantly higher in patients without malnutrition, this relation was insignificant based on NRN-2002. No significant difference was observed between malnourished and well-nourished groups for all tools regarding BF .Anthropometric variables of patients according to the GLIM, NRS-2002, and ESPEN criteria are summarized in The detailed results of the sensitivity, specificity, positive and negative predictive values for all patients and according to sex, type of leukemia, CR, and risk of disease are available in p < 0.001), while the agreement between GLIM and ESPEN was fair . Moreover, when comparing the ESPEN with NRS-2002 as a reference tool, ESPEN showed a sensitivity of 93.55%, while the specificity was 22.86%. The agreement between these two instruments was fair . We also found a moderate agreement between GLIM, NRS-2002, and ESPEN   .p < 0.001); however, the agreement between GLIM and ESPEN , and between ESPEN and NRS-2002  were fair. We also found a moderate agreement between GLIM, NRS-2002, and ESPEN . In female patients, the agreement between GLIM and NRS-2002 was substantial , and the agreement between GLIM and ESPEN was moderate . However, the agreement between ESPEN and NRS-2002 was fair . We also found a moderate agreement between GLIM, NRS-2002, and ESPEN   .p < 0.001), while the agreement between GLIM and ESPEN was slight . There was no significant agreement between ESPEN and NRS-2002. We observed a fair deal between these three tools . In ALL patients, our results indicated substantial agreement between GLIM and NRS-2002  and moderate agreement between GLIM and ESPEN . Nevertheless, there was a fair agreement between ESPEN and NRS-2002 . Also, the agreement between these three tools was moderate   .p < 0.001), while the agreement between GLIM and ESPEN  and between ESPEN and NRS-2002  were fair. We found a moderate agreement between all three mentioned tools . In patients who achieved a second CR, the agreement between GLIM and NRS-2002 was substantial . The agreement between GLIM, ESPEN, and NRS-2002 was not significant. In patients in the third CR, the agreement between GLIM and NRS-2002 was perfect , while the agreement between GLIM and ESPEN and between ESPEN and NRS-2002 were insignificant. We found a substantial agreement between these three tools   .p = 0.003). In patients with adverse risk of disease, the agreement between GLIM and NRS-2002 was substantial ; however, the agreement between GLIM and ESPEN  and ESPEN and NRS-2002  were fair and slight, respectively. We also found a moderate agreement between these three tools   .This study is one of the first to compare different tools for evaluating malnutrition in patients with leukemia who are also candidates for allo-HSCT. According to NRS-2002, GLIM, and ESPEN criteria, the results showed that 33.6, 26, and 8.6% of the patients were malnourished, respectively. Regardless of gender, type of leukemia, CR status, and risk status, GLIM and NRS-2002 instruments had a perfect agreement in diagnosing malnutrition; however, the ESPEN criterion showed fair agreement with the other two instruments (GLIM and NRS-2002). In all patients, we obtained a moderate agreement for all tools. In general, these results were maintained in subgroup analyses.Although malnutrition caused by disease is known as a serious problem, a gold standard for its diagnosis has not been introduced so far. While Peng et al. introduced the NRS-2002 as the first choice for assessing malnutrition before HSCT , studiesMost previous studies evaluated malnutrition in patients with solid tumors, and studies in HSCT candidate patients are limited. However, the nutritional status of patients with hematologic malignancies is not vastly different from general oncology patients . VariousThe prevalence of malnutrition in this study based on the GLIM criteria was 26%, which was in line with previous studies in cancer patients. Previous studies have reported that the rate of malnutrition was 32% in head and neck cancer , 24% in In this study, our choice of comparing these tools stemmed from a prior investigation that pitted the NRS-2002 against the SGA. The findings revealed a high degree of concordance, with NRS-2002 exhibiting a specificity of 94.81% in identifying severe malnutrition. This strong agreement might be attributed to NRS-2002’s balanced consideration of both nutritional status and disease severity. It implies that NRS-2002 could categorize patients as high-risk primarily due to the severity of their illness .Hence, the primary objective of our study was to assess NRS-2002 against alternative tools like the GLIM criteria, focusing on their specificity and sensitivity. While the PG-SGA is widely regarded as the gold standard for cancer patients, our study specifically aimed to compare screening and assessment tools commonly used in the pre-transplant setting. The choice of which method to employ depends on factors such as available infrastructure, resources, automation feasibility, and the healthcare environment .Moreover, previous research has indicated that GLIM criteria demonstrate ‘moderate agreement’ (kappa = 0.426) when compared to SGA. This suggests that GLIM criteria can effectively assess the nutritional status of cancer patients even without considering SGA, a recommendation supported by existing literature , 39. AddThe results of our study determined that the GLIM and ESPEN criteria have a fair agreement in diagnosing malnutrition in leukemia patients who are candidates for HSCT. In the subgroup analysis, the agreement ranged from weak to moderate; however, in the group of patients with favorable risk at the beginning of transplantation, a perfect agreement was shown, but because only three patients were in that group, the clinical significance of the result is reduced, and further studies are needed. The results of our study were consistent with previous studies; for example, Ruiz et al. determined the prevalence of malnutrition in outpatient cancer patients based on the GLIM criteria, 46.7%, and based on the ESPEN criteria, 17.6% . The kapOne of the important reasons for the difference in the prevalence of malnutrition obtained with the above two tools is the reduction of the BMI cut-off required to diagnose malnutrition in the ESPEN criteria. In addition to all the parameters for diagnosing malnutrition based on ESPEN, the GLIM criterion also has a series of etiological criteria that can be effective in the high prevalence of malnutrition compared to the ESPEN criterion.The NRS-2002 and GLIM criteria perfectly agreed in diagnosing malnutrition in this study. In the subgroup analysis, the agreement ranged from substantial to perfect. 33.6% of patients in the current research based on NRS-2002 were recognized as being at nutritional risk, comparable to previous studies in patients with cancer and candidates for HSCT , 20. AmoMalnutrition assessment is important because its early diagnosis leads to better and more effective interventions in the hospital. Early intervention prevents the progression of malnutrition to cancer cachexia . VariousOne of the strengths of this study was that the total population was patients with leukemia who were candidates for allo-HSCT, which had low heterogeneity. The number of participants in the project was greater than in similar studies because other hematological malignancies and autologous HSCT candidates were omitted. Additionally, three separate days were used for the collection of dietary data. As a result, individual variation could more effectively be detected. Despite the strengths, there were also limitations. The cross-sectional design makes the cause-and-effect relationship between the diagnosis of malnutrition and post-transplant outcomes unclear. Considering the differences in ethnicity within other studies and its effect on some parameters for determining malnutrition, such as BMI, FFMI and ASMI, it is necessary to conduct similar studies in different populations in different countries to obtain results with higher clinical significance. While our study aimed to compare the performance of three tools, namely NRS-2002, ESPEN criteria, and GLIM criteria, it is important to acknowledge that these tools serve distinct roles in the nutritional assessment process. NRS-2002 is a recognized screening tool designed to identify patients at risk of malnutrition , whereasThis study demonstrates that the GLIM criterion perfectly agreed with the NRS-2002. The ESPEN criterion had a fair agreement with other two instruments, and it has problems in early diagnosis of malnutrition, especially in the pre-cachexia stage. The agreement of the three tools with each other was also moderate. In the future, it is imperative to undertake rigorous scientific investigations focused on discerning the optimal diagnostic modality for malnutrition and fostering consensus in this regard.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving humans were approved by Tehran University of Medical Sciences ethics committee. The studies were conducted in accordance with the local legislation and institutional requirements. The participants provided their written informed consent to participate in this study.RK and HM designed the study. SM, MB, and AR did data collection. ES did the analysis. HI, SZ-M, and RK wrote the article. SW provided additional scientific analysis and reviewed and edited the manuscript. Finally, HM and SW carefully read the text and tables and made the necessary edits. All authors contributed to the article and approved the submitted version.