Two thousand five hundred and sixty swab samples were collected from December 2022 to December 2023 in China.PCR was used to detect FBoV and amplify its VP2 and NS1 gene cod-ing equences,and bioinformatics was used to analyze the genetic diversity of FBoV.The results showed that the total positive rate of FBoV was 4.6％(119/2 560).Genetic variation analysis showed that FBoV existed in a variety of genotypes,and FBOV-1 was the main epidemic type in China.The 15 FBoV-1 strains,four FBoV-2 strains and one FBoV-3 strains identified in this study were genetically close to the strains identified in China,the United States,Thailand,Australia and Portugal.Sequence analysis showed that the identities of amino acid sequence of NS1 and VP2 genes between the sequenced strains and the reference strains were 59.13％-99.25％and 96.41％-100.00％,respectively.The amino acid identities of NS1 and VP2 among the newly sequenced FBoV strains were 60.00％-100.00％and 96.41％-100.00％,respectively,which indicated that the FBov strains circulating in China had great genetic diversity.This study enriched the data for elucidating the epidemic status of FBoV in China,and provided the basis for the subsequent diag-nosis,prevention and control of FBoV.

Objective: To investigate the regulatory effect of Jieduan Niwan Formula (JDNWF) drug-containing serum on AMPK-mediated mitochondrial quality control in D-GalN-induced HepG2 cells. Methods: Twenty male Wistar rats were randomly divided into blank control and JDNWF-containing serum groups, 10 rats per group. The JDNWF-containing serum group was gavaged with JDNWF (21.7 g/kg), whereas the blank control group was gavaged with saline. Blood was collected to prepare JDNWF-containing and blank control serum. Cell viability, mitochondrial damage indicators, and MQC pathway protein expression levels were evaluated to determine the optimal volume fraction of JDNWF. HepG2 cells were divided into control, D-GalN, DMSO, AMPK inhibitor, JDNWF drug-containing serum, and JDNWF drug-containing serum plus AMPK inhibitor groups, and corresponding drug interventions were administered to each group. Cells were collected after the interventions, and the CCK-8 assay was used to measure cell viability, the 2′-7′-dichlorodihydrofluorescein diacetate fluorescent probe was used to detect reactive oxygen species (ROS) levels, JC-1 was used to detect mitochondrial membrane potential, thiobarbituric acid was used to measure malondialdehyde (MDA) levels, WST-8 was used to measure superoxide dismutase (SOD) activity, and western blotting was used to detect the expression levels of mitochondrial quality control-related proteins, including p-AMPK, AMPK, PGC-1α, NRF1, TFAM, MFN2, and DRP1. Results: 5% JDNWF drug-containing serum most significantly restored cell viability, mitochondrial damage markers, and MQC pathway protein expression in the model group. Therefore, it was chosen for intervention in subsequent experiments. Compared to the control group, the cell viability of the D-GalN, DMSO, and AMPK inhibitor groups was significantly reduced (P<0.01). In contrast, the heterogeneity of mitochondrial membrane potential, ROS, and MDA levels was significantly increased (P<0.01), and SOD activity was significantly decreased (P<0.01). The p-AMPK, PGC-1α, NRF1, TFAM, MFN2, and DRP1 protein expression levels were significantly decreased (P<0.01). After JDNWF drug-containing serum intervention, compared to the DMSO group, cell viability significantly increased (P<0.01), mitochondrial membrane potential heterogeneity, ROS, and MDA levels significantly decreased (P<0.01), SOD activity significantly increased (P<0.01), and p-AMPK, PGC-1α, NRF1, TFAM, and MFN2 protein expression levels significantly increased (P<0.01), whereas DRP1 protein expression significantly decreased (P<0.01). Compared to the JDNWF drug-containing serum group, the cell viability in the JDNWF plus AMPK inhibitor group significantly decreased (P<0.01), mitochondrial membrane potential heterogeneity and ROS levels significantly increased (P<0.01), MDA levels significantly increased (P<0.05), SOD activity significantly decreased (P<0.05), p-AMPK, PGC-1α, NRF1, and TFAM protein expression levels significantly decreased (P<0.01), MFN2 protein expression significantly decreased (P<0.05), and DRP1 protein expression significantly increased (P<0.01). Conclusion JDNWF drug-containing serum may restore mitochondrial function and improve D-GalN-induced HepG2 cell injury by regulating AMPK-mediated mitochondrial quality control.

Objective　To detect clustered regularly interspaced short palindromic repeats (CRISPR) in Listeria monocytogenes, and analyze the structure and homology of CRISPR loci. Methods Totally 34 strains of Listeria monocytogenes isolated in our laboratory were identified, PCR amplified and sequenced. The repeat sequence structure and spacer sequence homology in CRISPR loci were analyzed by bioinformatics software. Results A total of 7 CRISPR loci were detected in 34 strains. The mutation rate of the first 2 and last 2 bases of the Repeat sequence of CRISPR loci was higher, while the mutation rate of the middle part was lower. Seven CRISPR sites form eight CRISPR structural types, among which the Repeat sequences of CRISPR1 and CRISPR2 are relatively conserved, while the Repeat sequences of CRISPR1 and CRISPR5 can form dumbbell shaped secondary structures. The number of Spacer sequences contained in each CRISPR site ranges from 2 to 15, with an average of 2.43. The 136 Spacer sequences detected were not only homologous to Listeria plasmids and bacteriophages, but also homologous to uncultured virus sequences, staphylococcal bacteriophages, and Listeria innocua. The same CRISPR genotype did not show large-scale clustering, but some strains in the same year were in the same evolutionary cluster with close genetic relationships. Conclusion The CRISPR structure of Listeria monocytogenes in this study exhibits high specificity, and its homology with bacteriophages provides a theoretical basis for the application of bacteriophages in the control and prevention of Listeria monocytogenes.

Two thousand five hundred and sixty swab samples were collected from December 2022 to December 2023 in China.PCR was used to detect FBoV and amplify its VP2 and NS1 gene cod-ing equences,and bioinformatics was used to analyze the genetic diversity of FBoV.The results showed that the total positive rate of FBoV was 4.6％(119/2 560).Genetic variation analysis showed that FBoV existed in a variety of genotypes,and FBOV-1 was the main epidemic type in China.The 15 FBoV-1 strains,four FBoV-2 strains and one FBoV-3 strains identified in this study were genetically close to the strains identified in China,the United States,Thailand,Australia and Portugal.Sequence analysis showed that the identities of amino acid sequence of NS1 and VP2 genes between the sequenced strains and the reference strains were 59.13％-99.25％and 96.41％-100.00％,respectively.The amino acid identities of NS1 and VP2 among the newly sequenced FBoV strains were 60.00％-100.00％and 96.41％-100.00％,respectively,which indicated that the FBov strains circulating in China had great genetic diversity.This study enriched the data for elucidating the epidemic status of FBoV in China,and provided the basis for the subsequent diag-nosis,prevention and control of FBoV.

AIM:To analyze and screen the risk factors for acute kidney injury(AKI)following heart transplantation(HT),and to establish a dynamic no-mograms prediction model to forecast early AKI af-ter HT.METHODS:A retrospective analysis was con-ducted on clinical data from HT recipients at Nan-jing First Hospital from October 2012 to June 2024.Patients were divided into AKI and non-AKI groups based on AKI occurrence within 7 days post-sur-gery,with a 8:2 ratio for training and testing sets.Lasso regression and multivariable logistic regres-sion were used to select influencing factors.A dy-namic nomogram model was visualized using R.In-ternal validation was performed using 1 000 boot-strap samples.Model accuracy and discrimination were evaluated using the area under the receiver operating characteristic curve(AUC-ROC),calibra-tion curves,and the Hosmer-Lemeshow goodness-of-fit test.The nomogram model was compared with the Cleveland score.RESULTS:The results of a multivariable logistic regression indicate that a his-tory of atrial fibrillation(OR=9.647,95％CI=1.961-47.470),vasoactive inotropic score(OR=1.094,95％CI=1.012-1.183),intraoperative transfusion of red blood cells or plasma(OR=10.200,95％CI=1.727-60.238),postoperative central venous pressure(OR=1.548,95％CI=1.186-2.021),and postoperative use of vancomycin(OR=25.082,95％CI=2.122-296.417)are independent risk factors for HT-AKI.The dynamic nomogram model achieved an AUC of 0.842(95％CI:0.676-0.971)in the test set,with a calibration plot showing a slope close to 1 and a Brier score of 0.173.The Hosmer-Lemeshow good-ness-of-fit test(x2=5.658,P=0.685)suggests good predictive performance of the model.Moreover,this model demonstrates superior discriminative ability compared to the Cleveland score.CONCLU-SION:This study identified preoperative,intraoper-ative,and postoperative risk factors influencing the occurrence of HT-AKI.The developed dynamic no-mogram model accurately identifies high-risk indi-viduals for early HT-AKI and is convenient for clini-cal use.

BACKGROUND: 3D-printing is widely used in regenerative medicine and is expected to achieve vaginal morphological restoration and true functional reconstruction. Mesenchymal stem cells-derived exosomes (MSCs-Exos) were applyed in the regeneration of various tissues. The current study aimed to explore the effctive of MSCs-Exos in vaginal reconstruction. METHODS: In this work, hydrogel was designed using decellularized extracellular matrix (dECM) and gelatin methacrylate (GelMA) and silk fibroin (SF). The biological scaffolds were constructed using desktop-stereolithography.The physicochemical properties of the hydrogels were evaluated; Some experiments have been conducted to evaluate exosomes’ effect of promotion vaginal reconstruction and to explore the mechanism in this process. RESULTS: It was observed that the sustained release property of exosomes in the hydrogel both in vitro and in vitro.The results revealed that 3D scaffold encapsulating exosomes expressed significant effects on the vascularization and musule regeneration of the regenerative vagina tissue. Also, MSCs-Exos strongly promoted vascularization in the vaginal reconstruction of rats, which may through the PI3K/AKT signaling pathway. CONCLUSION The use of exosome-hydrogel composites improved the epithelial regeneration of vaginal tissue, increased angiogenesis, and promoted smooth muscle tissue regeneration. 3D-printed, lumenal scaffold encapsulating exosomes might be used as a cell-free alternative treatment strategy for vaginal reconstruction.

The use of nitrification inhibitors has been suggested as a strategy to decrease cadmium(Cd)accumulation in crops.However,the most efficient nitrification inhibitor for mitigating crop Cd accumulation remains to be elucidated,and whether and how changes in soil microbial structure are involved in this process also remains unclear.To address these questions,this study applied three commercial nitrification inhibitors,namely,dicyandiamide(DCD),3,4-dimethylpyrazole phosphate(DMPP),and nitrapyrin(NP),to pakchoi.The results showed that both DCD and DMPP(but not NP)could efficiently decrease Cd concentrations in pakchoi in urea-and ammonium-fertilized soils.In addition,among the three tested nitrification inhibitors,DMPP was the most efficient in decreasing the Cd concentration in pakchoi.The nitrification inhibitors decreased pakchoi Cd concentrations by suppressing acidification-induced Cd availability and reshaping the soil microbial structure;the most effective nitrification inhibitor was DMPP.Ammonia oxidation generates the most protons during nitrification and is inhibited by nitrification inhibitors.Changes in environmental factors and predatory bacterial abundance caused by the nitrification inhibitors changed the soil microbial structure and increased the potential participants in plant Cd accumulation.In summary,our study identified DMPP as the most efficient nitrification inhibitor for mitigating crop Cd contamination and observed that the soil microbial structural changes caused by the nitrification inhibitors contributed to decreasing Cd concentration in pakchoi.

AIM:To analyze and screen the risk factors for acute kidney injury(AKI)following heart transplantation(HT),and to establish a dynamic no-mograms prediction model to forecast early AKI af-ter HT.METHODS:A retrospective analysis was con-ducted on clinical data from HT recipients at Nan-jing First Hospital from October 2012 to June 2024.Patients were divided into AKI and non-AKI groups based on AKI occurrence within 7 days post-sur-gery,with a 8:2 ratio for training and testing sets.Lasso regression and multivariable logistic regres-sion were used to select influencing factors.A dy-namic nomogram model was visualized using R.In-ternal validation was performed using 1 000 boot-strap samples.Model accuracy and discrimination were evaluated using the area under the receiver operating characteristic curve(AUC-ROC),calibra-tion curves,and the Hosmer-Lemeshow goodness-of-fit test.The nomogram model was compared with the Cleveland score.RESULTS:The results of a multivariable logistic regression indicate that a his-tory of atrial fibrillation(OR=9.647,95％CI=1.961-47.470),vasoactive inotropic score(OR=1.094,95％CI=1.012-1.183),intraoperative transfusion of red blood cells or plasma(OR=10.200,95％CI=1.727-60.238),postoperative central venous pressure(OR=1.548,95％CI=1.186-2.021),and postoperative use of vancomycin(OR=25.082,95％CI=2.122-296.417)are independent risk factors for HT-AKI.The dynamic nomogram model achieved an AUC of 0.842(95％CI:0.676-0.971)in the test set,with a calibration plot showing a slope close to 1 and a Brier score of 0.173.The Hosmer-Lemeshow good-ness-of-fit test(x2=5.658,P=0.685)suggests good predictive performance of the model.Moreover,this model demonstrates superior discriminative ability compared to the Cleveland score.CONCLU-SION:This study identified preoperative,intraoper-ative,and postoperative risk factors influencing the occurrence of HT-AKI.The developed dynamic no-mogram model accurately identifies high-risk indi-viduals for early HT-AKI and is convenient for clini-cal use.

The importance of structural variants(SVs)for human phenotypes and diseases is now recognized.Although a variety of SV detection platforms and strategies that vary in sensitivity and specificity have been developed,few benchmarking procedures are available to confidently assess their performances in biological and clinical research.To facilitate the validation and application of these SV detection approaches,we established an Asian reference material by characterizing the genome of an Epstein-Barr virus(EBV)-immortalized B lymphocyte line along with identified benchmark regions and high-confidence SV calls.We established a high-confidence SV callset with 8938 SVs by integrating four alignment-based SV callers,including 109x Pacific Biosciences(PacBio)continuous long reads(CLRs),22 x PacBio circular consensus sequencing(CCS)reads,104x Oxford Nanopore Technologies(ONT)long reads,and 114×Bionano optical mapping plat-form,and one de novo assembly-based SV caller using CCS reads.A total of 544 randomly selected SVs were validated by PCR amplification and Sanger sequencing,demonstrating the robustness of our SV calls.Combining trio-binning-based haplotype assemblies,we established an SV benchmark for identifying false negatives and false positives by constructing the continuous high-confidence regions(CHCRs),which covered 1.46 gigabase pairs(Gb)and 6882 SVs supported by at least one diploid haplotype assembly.Establishing high-confidence SV calls for a benchmark sample that has been characterized by multiple technologies provides a valuable resource for investigating SVs in human biology,disease,and clinical research.

Objective:To explore the value of proton density fat fraction(PDFF) based on histogram analysis for quantification hepatic steatosis and fibrosis in rabbit model and the interference of hepatic fibrosis to the evaluation of hepatic steatosis with PDFF.Methods:From March to November 2020, 135 New Zealand white rabbits were randomly divided into control group ( n=30) and experimental group ( n=105) using a random number table. The volume ratio of CCl 4 and olive oil was 1∶1 to prepare 50% CCl 4 oil solution, and experimental rabbits were subcutaneously injected with the oil solution. An equal dose of normal saline was subcutaneously injected for control group rabbits. At the end of the 4 th, 8 th, and 12 th week, 35 in the experimental group and 10 rabbits in the control group were randomly selected to conduct the mDixon-Quant scanning, and histogram analysis of PDFF was analyzed including volume, mean, median, standard deviation, 25 th, 50 th, 75 th, 90 th quantile, skewness, kurtosis, entropy and inhomogeneity. After the examination, the rabbits were sacrificed and the liver percentage of steatosis (PSH) and fibrosis (POF) were recorded by semi-quantitative analysis. Spearman correlation analysis was used to correlate PDFF with PSH and POF. Multiple linear regression analysis was used to determine independent PDFF histogram parameters for evaluating PSH and POF. A receiver operator characteristic (ROC) curve was used to assess the diagnostic accuracy of PDFF for discriminating mild from moderate-severe hepatic steatosis and mild from moderate-severe hepatic fibrosis with median of PSH or POF for dichotomy, and DeLong test was used to compare the area under the curve (AUC). With the correction of hepatic fibrosis, correlation coefficient and AUC were compared of PDFF for discrimination mild from moderate-severe hepatic steatosis. Results:The PDFF mean, median, standard deviation, 75 th, 90 th showed correlation with PSH ( r=0.558, 0.522, 0.319, 0.723, 0.646, -0.589, all P<0.05). The entropy and 75 th were independent parameters for evaluating PSH (β=2.347, -5.960, P=0.018, 0.001). The PDFF 75 th was the optimal parameter for discriminating mild from moderate-severe hepatic steatosis with AUC=0.915 ( P=0.001). The PDFF volume, mean, median, standard deviation, 75 th, 90 th, entropy showed correlation with POF ( r=0.355, 0.393, 0.376, 0.298, 0.485, 0.426, -0.681, all P<0.05). The entropy, standard deviation and volume (β=-11.041, 1.356, 0.190, P=0.001, 0.026, 0.016) were independent parameters for evaluation of hepatic fibrosis, and the entropy was the optimal parameter for hepatic fibrosis (AUC=0.771, P=0.001). The correlation between PSH and PDFF 75 th was less pronounced when fibrosis was present ( r=0.512, P=0.001) than when fibrosis was absent ( r=0.751, P=0.002). The PDFF 75 th showed a significant difference in discriminating mild hepatic steatosis from moderate-severe hepatic steatosis after correction of POF (AUC=0.895, 0.950, Z=2.970, P=0.025). Conclusions:PDFF based on histogram analysis provided a noninvasive, accurate estimation of quantification for hepatic steatosis and fibrosis. Hepatic fibrosis reduced the correlation between hepatic steatosis and PDFF and the presence of hepatic fibrosis can confound the quantification of hepatic steatosis with PDFF.