ObjectiveTo explore the effects of serine hydroxymethyltransferase 1 （SHMT1） rs1979277 polymorphisms on pharmacokinetic characteristics and clinical prognosis of methotrexate （MTX） in children with acute lymphoblastic leukemia （ALL）. MethodsMatrix-assisted laser desorption/ionization time of flight mass spectrometry was used for SHMT1 rs1979277 genotyping in children with ALL . Clinical data including serum MTX concentrations， incidences of adverse events， and ALL relapse after chemotherapy with MTX were collected. The associations of SHMT1 rs1979277 G>A genotypes with dose-adjusted serum concentrations （C/D ratios）， adverse events of MTX， and relapse were analyzed. The associations between rs1979277 genotypes and SHMT1 expression were explored based on Bioinformatics methods. ResultsAmong the 146 children with ALL included， the rs1979277 GG homozygous genotype accounted for 85.62% （125/146）， while the GA heterozygous genotype accounted for 14.38% （21/146）. The frequency of the G allele was 92.81% （271/292）， while the A allele was only 7.19% （21/292）. Children with the GG homozygous genotype had higher median C/D ratios of MTX in 24 h ［12.06 （μmol·m²）/（L·g）］ and higher relapse rates （12.80%） than those in GA heterozygous genotype carriers ［10.96 （μmol·m²）/（L·g）， and 9.52%， respectively］. However， none of the above differences were statistically significant （all P>0.05）. The incidences of respiratory （19.05%） and liver disorders （33.33%） in children with the GA heterozygous genotype were significantly higher than those in GG homozygous genotype carriers （4.00% and 12.00%， respectively， P<0.05）. There were no statistically significant differences in the incidences of other adverse events. Bioinformatics analysis showed that the rs1979277 A allele was significantly associated with higher SHMT1 expression in multiple tissues， such as the tibial artery， pancreas， and adrenal gland （P<0.05）. ConclusionSHMT1 rs1979277 GA genotype may be a risk factor for respiratory and liver disorders in ALL children treated with MTX.

ObjectiveTo explore the effects of vascular endothelial growth factor A （VEGFA） rs2010963 and rs3025039 polymorphisms on chemotherapy toxicities and clinical prognosis in children with brain tumors. MethodsA total of 104 pediatric patients with brain tumors receiving standardized chemotherapy were enrolled. Matrix-assisted laser desorption/ionization time of flight mass spectrometry was used for VEGFA rs2010963 and rs3025039 genotyping. The χ² test was applied to analyze the association between genotypes and chemotherapy‑related toxicities. Cox regression was used to evaluate the correlations of clinicopathological characteristics and genotypes with the progression‑free survival （PFS）. In addition， bioinformatic analyses were conducted to investigate the regulatory factors potentially affected by the two SNP loci. ResultsThe VEGFA expression in brain tumors （5.17±1.81） was significantly higher than that in normal tissues （4.33±1.56， P<0.001）. Patients with high VEGFA expression had significantly worse overall survival than patients with low VEGFA expression （P<0.001）. Among the 104 children with brain tumors included， the rs2010963 CC， CG， and GG genotypes accounted for 14.42%， 55.77%， and 29.81%， respectively. The frequencies of C and G alleles were 42.31% and 57.69%， respectively. The rs3025039 CC， CT， and TT genotypes accounted for 70.19%， 25.96%， and 3.85%， respectively. The frequencies of C and T alleles were 83.17% and 16.83%， respectively. The children with the rs3025039 CC genotype had significantly higher incidences of thrombocytopenia （46.58%） and gastrointestinal toxicity （56.16%） than CT genotype carriers （22.22% and 33.33%， respectively， P<0.05）， and significantly lower incidences of coagulation disorders （4.11%） than TT genotype carriers （50.00%， P<0.05）.The incidence of hyperlipidemia （2.74%） was significantly lower than that in the CT genotype carriers （14.82%， P<0.05）. The tumor type and the rs2010963 genotype were significantly associated with PFS （P<0.05） in univariable and multivariable Cox regression analysis. Bioinformatic analysis indicated that the rs2010963 and rs3025039 polymorphisms regulated VEGFA expression by affecting the binding of transcription factors and miRNAs to their target gene sequences， respectively. ConclusionThe VEGFA rs3025039 CC genotype is a risk factor for thrombocytopenia and gastrointestinal toxicity， and a protective factor for coagulation disorders and hyperlipidemia. The rs2010963 CG genotype is a protective factor for brain tumor progression.

Objective To translate Evidence-Informed Decision-Making Competence Measure for Chinese nurses and test its validity and reliability.Methods A research group was set up to use the Brislin translation model to translate the original scale into Chinese,and the back translation,cross-cultural adaptation,pre-experiment and cognitive interview were conducted to finally form the Chinese version of the Evidence-Informed Decision-Making Competence Measure for nurses.A total of 1 247 nurses from 7 tertiary A hospitals in Beijing,Hubei,Hunan and Xinjiang were selected by convenience sampling method in April 2024 to test its reliability and validity.Results 1 026 effective question-naires were collected,with an effective recovery rate of 82.28％.The Chinese version of the Evidence-Informed Decision-Making Competence Measure included 25 items,including knowledge/skill,attitude and behavior.A total of 3 common factors were extracted from exploratory factor analysis,and the cumulative variance contribution rate was 91.725％.The content validity index at the item level was 0.83-1.00;the content validity index at the scale level was 0.988;the calibration association validity was 0.496.The Cronbach's α coefficient of the whole scale was 0.992;the half-point reliability was 0.930;the retest reliability was 0.927.Conclusion The Chinese version of Evidence-Informed Decision-Making Competence Measure for nurses has good reliability and validity,and it can be used to evaluate the evidence-informed decision-making competence of Chinese nurses,provide references for promoting evidence-based nursing practice and evidence-informed decision-making.

Gene synthesis is an enabling technology that supports the development of synthetic biology. The existing approaches for de novo gene synthesis generally have tedious operation, low efficiency, high error rates, and limited product lengths, being difficult to support the huge demand of synthetic biology. The assembly and error correction are the keys in gene synthesis. This study first designed the oligonucleotide sequences by reasonably splitting the virus genome of approximately 10 kb by balancing the parameters of sequence design software ability, PCR amplification ability, and assembly enzyme assembly ability. Then, two-step PCR was performed with high-fidelity polymerase to complete the de novo synthesis of 3.0 kb DNA fragments, and error correction reactions were performed with T7 endonuclease Ⅰ for the products from different stages of PCR. Finally, the virus genome was assembled by 3.0 kb DNA fragments from de novo synthesis and error correction and then sequenced. The experimental results showed that the proposed method successfully produced the DNA fragment of about 10 kb and reduced the probability of large fragment mutations during the assembly process, with the lowest error rate reaching 0.36 errors/kb. In summary, this study developed an efficient de novo method for synthesizing a viral genome of about 10 kb with T7 endonuclease Ⅰ-mediated error correction. This method enabled the synthesis of a 10 kb viral genome in one day and the correct plasmid of the viral genome in five days. This study optimized the de novo gene synthesis process, reduced the error rate, simplified the synthesis and assembly steps, and reduced the cost of viral genome assembly.
Genome, Viral/genetics* ; Polymerase Chain Reaction/methods* ; DNA, Viral/genetics* ; Bacteriophage T7/enzymology* ; Synthetic Biology/methods*

Bacillus is a class of spore-producing Gram-positive bacteria that produce a variety of antimicrobial substances with different structures and functions. The application of the antimicrobial substances produced by Bacillus can effectively inhibit the activity of harmful bacteria and fungi and promote the sustainable development of green agriculture. The antimicrobial substances produced by Bacillus mainly include proteins, lipopeptides, polyketones, and polypeptides. This paper reviews the synthesis gene clusters, synthesis pathways, structures, and mechanisms of various antimicrobial substances produced by Bacillus and discusses the challenges in the industrial application of these antimicrobial substances. Furthermore, this paper clarifies the future research and development focuses and prospects the application prospects, and provides comprehensive theoretical support for the in-depth research and wide application of the antimicrobial substances produced by Bacillus.
Bacillus/genetics* ; Anti-Infective Agents/metabolism* ; Bacterial Proteins/genetics* ; Antimicrobial Peptides/biosynthesis* ; Lipopeptides/biosynthesis*

Bacillus velezensis DJ1 exhibits broad-spectrum antagonistic activity against diverse phytopathogenic fungi, while its biocontrol mechanisms against Fusarium graminearum, the causal agent of maize stalk rot, remain poorly characterized. In this study, we integrated genomics and transcriptomics to elucidate the antifungal mechanisms of strain DJ1. The results demonstrated that DJ1 inhibited F. graminearum with the efficacy of 64.4%, while its polyketide crude extract achieved the control efficacy of 55% in pot experiments against this disease. Whole-genome sequencing revealed a single circular chromosome (3 929 792 bp, GC content of 47%) harboring 12 biosynthetic gene clusters for secondary metabolites, six of which encoded known antimicrobial compounds (macrolactin H, bacillaene, difficidin, surfactin, fengycin, and bacilysin). Transcriptomic analysis identified 243 differentially expressed genes (152 upregulated and 91 downregulated, P < 0.05), which were potentially associated with the antagonistic activity against F. graminearum. KEGG enrichment analysis highlighted activation (P < 0.05) of cysteine/methionine metabolism, pentose phosphate pathway, and polyketide biosynthesis pathways, indicating that DJ1 employed synergistic strategies involving antimicrobial compound synthesis, energy metabolism enhancement, and nutrient competition to suppress pathogens. This study provides a theoretical foundation for developing novel microbial resources and application technologies to combat phytopathogenic fungi.
Fusarium/drug effects* ; Bacillus/metabolism* ; Plant Diseases/prevention & control* ; Antifungal Agents/pharmacology* ; Genomics ; Zea mays/microbiology* ; Transcriptome ; Gene Expression Profiling ; Antibiosis ; Multigene Family ; Multiomics

Objective:This study aimed to explore the molecular mechanisms of the maladaptation of H3N2 influenza virus in chicken embryos, provide a theoretical basis for the restoration of H3N2 influenza vaccine production in chicken embryos.Methods:Samples of respiratory secretions from patients with influenza-like symptoms (Influenza-like Illness, ILI) caused by H3N2 influenza virus were inoculated into chicken embryos and Madin-Darby Canine Kidney cells (MDCK), respectively. After isolating the virus, hemagglutination experiments were conducted to detect hemagglutination titers and hemagglutination inhibition experiments were used to compare antigenic differences; further, whole-genome sequencing of H3N2 influenza virus was performed using second-generation high-throughput gene sequencing (Next Generation High-Throughput Gene Sequencing, NGS), and key amino acid sites of mutations were identified through sequence alignment; combined with sialic acid receptor binding experiments, the differences in the binding of wild-type and mutant receptor binding sites (RBS) to sialic acid receptors were compared; finally, molecular docking and molecular dynamics simulation method were used to explore the specific molecular mechanisms of how mutation sites affect the differences in the affinity of the RBS pocket for sialic acid receptors.Results:The hemagglutination assay result indicated that both chicken embryos and MDCK cells could isolate the influenza virus, and the hemagglutination inhibition test showed that no antigenic differences were produced in the isolated strains. NGS analysis revealed that the H3N2 virus underwent an F195Y mutation in the (RBS) region of the hemagglutinin (HA) protein after adaptation through chicken embryo passages. Receptor-binding experiments demonstrated that the F195Y mutation enhanced the virus′s binding ability to α2, 3-linked sialic acid glycan (Neu5Acα2-3Galβ1-4GlcNAcβ-PAA, 3′SLN), while the mutation did not affect the affinity of the RBS pocket for α2, 6-linked sialic acid glycan (Neu5Acα2-6Galβ1-4GlcNAcβ-PAA, 6′SLN). Molecular docking and molecular dynamics simulation result indicate that the F195Y mutation, by replacing a hydrophobic amino acid with a hydrophilic one, leads to a significant decrease in the structure of the RBS pocket, enhancing the binding stability of the H3N2 influenza virus with α2, 3-sln. This is specifically manifested by an increase in binding time and an increase in the number of hydrogen bonds at the RBS site with the receptor. Furthermore, the F195Y mutation does not alter the binding of the virus to other receptors.Conclusions:The F195Y mutation in the RBS pocket of H3N2 influenza virus is a key site affecting the viral chicken embryo inadaptability.

Objective:To explore the influences of FibroScan detection on the pregnancy outcomes of pregnant rats transfected with hepatitis B virus and the growth of their offspring rats.Methods:Sixty SPF-grade SD rats with a male-to-female ratio of 5∶1 were selected，randomly divided the female rats into the groups of mid-pregnancy with hepatitis B，late-pregnancy with hepatitis B，normal mid-pregnancy，normal late-pregnancy，and control group，and the rats in the mid-pregnancy and late-pregnancy groups were subjected to FibroScan testing on the 10th and 15th days of gestation respectively. The number of offspring rats in each group was randomly reduced to 10 on the 3rd day after delivery，and the offspring rats were divided into the groups of mid-pregnancy with hepatitis B，late-pregnancy with hepatitis B，normal mid-pregnancy，normal late-pregnancy，and control group. The pregnancy outcome levels of pregnant rats were observed by adopting a stratified analysis strategy，including differences in weight changes，number of deliveries，fetal rat outcomes，lactation conditions，litter size，and litter weight. The growth levels of offspring rats，including differences in weight，body length，tail length，and the time of fur growth，tooth eruption，eye opening，and ear standing on the 21st day were observed，and the survival rate of the offspring rats was compared.Results:With regard to the pregnancy outcome levels of the pregnant rats，there were no statistically significant differences in weight changes during pregnancy among the five groups of pregnant rats［ F（4，45）=2.627，Adjusted P=0.222］. There were also no statistically significant differences in the number of deliveries，fetal rat outcomes，and lactation conditions（Adjusted P=1.000）. The number of deliveries［ F（4，21.095）=2.280，Adjusted P=0.222］and litter weight［ F（4，20.128）=2.159，Adjusted P=0.222］showed no statistically significant differences. After correction using the BH method，none of the indicators showed significant differences（ P>0.05）. In terms of the growth levels of the offspring rats，among the five groups of offspring rats，there were no statistically significant differences in body weight on the 21st day［H（4）=11.623，Adjusted P=0.135］，body length on the 21st day［H（4）=10.962，Adjusted P=0.135］，and tail length on the 21st day［H（4）=9.126，Adjusted P=0.058］. Besides，the differences in the time of fur growth，tooth eruption，eye opening，and ear standing［H（4）=0.000，Adjust P=1.000］showed no statistical significance. The survival rate on the 7th day，14th day，and 21st day was 100%，with no significant differences（Adjusted P=1.000）. After correction using the BH method，none of the indicators showed significant differences（ P>0.05）. Conclusion:FibroScan detection had no significant influences on the pregnancy outcomes of pregnant rats with hepatitis B or on the growth of their offspring rats in multiple stages，dimensions，and indicators，indicating that FibroScan detection is safe for pregnant rats. This research provides an animal experimental basis for the safe use of FibroScan in pregnant women with chronic HBV infection.

This study aims to analyze the screening results and epidemiological characteristics of prostate cancer (PCa) among elderly males in the rural areas of Songjiang, Shanghai City, through the implementation of a preliminary prostate-specific antigen (PSA) screening based on a community-linkage model, and to explore an effective screening approach. A retrospective observational study design was employed to collect data from residents who underwent PSA screening at Songjiang Hospital affiliated to Shanghai Jiao Tong University School of Medicine, in collaboration with multiple community health service centers in Songjiang District, Shanghai City, between June 2022 and June 2024, through free clinics and annual health examinations. Prostate biopsy was recommended for individuals with total PSA (tPSA) levels >10 ng/ml and those with 4 ng/ml≤tPSA≤10 ng/ml and abnormal free-to-total PSA (f/tPSA) ratios. Clinical characteristics of detected PCa patients were analyzed. Follow-up was conducted through phone calls and home visits by family doctors, coupled with enhanced health education. The results indicated that a total of 17 198 residents participated in the screening, among which 2 234 (12.99%) had tPSA levels between 4 ng/ml and 10 ng/ml, and 257 (1.49%) had tPSA levels >10 ng/ml. Ultimately, 417 residents underwent prostate biopsy, with 171 being diagnosed with PCa, yielding a positive biopsy rate of 41.00% and a PCa detection rate of 0.99%. The predominant pathological subtype among PCa patients was adenocarcinoma (168 cases, 98.24%). Of the 146 PCa patients who received treatment, the majority were classified as intermediate or high-risk (124 cases, 84.93%). Furthermore, with the optimization of the screening model, there was a significant increase in the proportion of subsequent outpatient visits. In conclusion, the community-linkage-based PSA screening model demonstrated high effectiveness in screening for PCa among elderly males in the rural areas of Songjiang, Shanghai City. Epidemiological findings revealed that PCa patients in this region are primarily composed of intermediate and high-risk groups, highlighting the need for intensified early screening and health education.

Objective:To investigate the efficacy and safety of sintilimab combined with the SOX regimen for adjuvant treatment of stage Ⅲgastric cancer after D2 radical resection and to provide a reference for individualized clinical treatment.Methods:The clinical data of 245 pa-tients with stage III gastric cancer who underwent D2 radical resection at the 940th Hospital of the Joint Support Force of the People's Liber-ation Army from June 2019 to May 2022 were retrospectively analyzed.The 180 patients who received only the SOX regimen were desig-nated the control group,and the 65 patients who received sintilimab combined with the SOX regimen were designated the experimental group.The 3-year disease-free survival(DFS)rate,overall survival(OS)rate,and adverse reactions among the two groups and different sub-groups(HER-2 positive,dMMR,CPS≥5)were compared.Results:The 3-year DFS(81.5%vs.59.4%)and OS(84.6%vs.70.6%)rates in the experimental group were significantly higher than those in the control group(both P<0.05).Group analysis showed that in patients with CPS≥5,the 3-year DFS(91.5%vs.67.0%)and OS(95.7%vs.71.6%)rates within the experimental group were significantly better than those in the control group(both P<0.05).Intra-group analysis within the experimental group showed that the 3-year DFS rate(91.5%vs.55.6%)and OS rate(95.7%vs.55.6%)of patients with CPS≥5 were significantly better than those of patients with CPS<5(both P<0.05).The overall and grade≥3 incidences of liver and kidney function damage,thyroid dysfunction,colitis,pneumonia,and rash in the experimental group were higher than those in the control group(all P<0.05),while the differences in other adverse reactions,including leukopenia were not statistic-ally significant(all P>0.05).Conclusions:Sintilimab combined with the SOX regimen can significantly improve 3-year DFS and OS rates in pa-tients with stage Ⅲ gastric cancer after surgery,especially in the CPS≥5 subgroup,with significant benefits and controllable safety.