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.

Myelin is an essential structure that facilitates rapid saltatory conduction in the nervous system. Discrepancies in myelin microstructure are a hallmark of numerous neurological disorders, rendering the assessment of myelin integrity and content an indispensable tool in clinical diagnostics and neuroscience research. Extensive research has been dedicated to scrutinizing its biochemical makeup and morphology under normal, pathological, and experimental conditions over the years. In this review, we present an updated summary of the myelin sheath's structure, composition, and developmental trajectory. We systematically enumerate and contrast eight prevalent myelin staining techniques across dimensions of sensitivity, specificity, and resolution, delving into their underlying staining principles. With an initial application of myelin histology on the mouse demyelination model, our review accentuates the accurate delineation of myelination and the microstructural analysis of the myelin sheath. Such insights are anticipated to significantly contribute to the evaluation and understanding of white matter pathologies.
Myelin Sheath/metabolism* ; Animals ; Humans ; Demyelinating Diseases/pathology* ; Staining and Labeling/methods*

OBJECTIVES: This study aimed to investigate the potential mediating role of plasma phosphorylated tau217 (p-tau217) in the association between periodontitis and mild cognitive impairment (MCI). METHODS: In this case-control study, patients diagnosed with MCI in the Neurology Department of the First Affiliated Hospital of Shandong Second Medical University from November 2023 to May 2024 were selected as the case group (MCI group). Cognitively normal (CN) volunteers, matched for age and education level and recruited from the physical examination center during the same period, served as the control group (CN group). The general demographic data of the study participants were collected. The Beijing versions of the Montreal Cognitive Assessment (MoCA), clinical dementia rating (CDR), and activities of daily living scale (ADL) were used to assess neuropsychological functions. Clinical periodontal examinations were conducted, the periodontal inflamed surface area (PISA) was calculated, and the periodontitis stage was determined in accordance with the 2018 classification. Fasting elbow venous blood samples were collected in the morning, and blood biochemical indicators were measured. Plasma p-tau217 levels were detected using enzyme-linked immunosorbent assay (ELISA). Statistical analyses were performed using t-test, Mann-Whitney U test, chi-square test, partial correlation analysis, multivariate Logistic regression analysis, multiple linear regression analysis, restricted cubic spline (RCS) regression analysis, and mediation effect analysis. RESULTS: Among the 192 participants, 96 belong to the MCI group and 96 to the CN group. The prevalence of periodontitis was 63.5% in the MCI group and 43.8% in the CN group, with a statistically significant difference (χ²=7.561, P=0.006). The plasma p-tau217 levels in the MCI group were significantly higher than those in the CN group [7.00 (4.27-9.65) ng/mL versus 2.02 (0.80-3.81) ng/mL, Z=-8.108, P<0.001]. Partial correlation analysis revealed that plasma p-tau217 levels were positively correlated with all the clinical periodontal indices (all P<0.001). After adjustments for baseline covariates, multivariate Logistic regression indicated that periodontitis was an independent risk factor for MCI. Patients with periodontitis had a 1.977-fold higher MCI risk than those without periodontitis (OR=1.977, 95%CI: 1.088-3.594, P=0.025). Moreover, the MCI risk for stage Ⅰ/Ⅱ periodontitis and stage Ⅲ/Ⅳ periodontitis was 1.878 times (OR=1.878, 95%CI: 1.029-3.425, P=0.040) and 2.625 times (OR=2.625, 95%CI: 1.073-6.246, P=0.035) higher than that for patients without periodontitis, respectively. Trend test showed that the MCI risk increased with periodontitis severity (P_trend=0.016). After adjustments for baseline covariates, multiple linear regression analysis showed that periodontitis was an independent risk factor for increased plasma p-tau217 levels (β=3.309, 95%CI: 2.363-4.254, P<0.001). Compared with patients without periodontitis, those with stage Ⅰ/Ⅱ periodontitis (β=1.838, 95%CI: 0.869-2.806, P<0.001) and stage Ⅲ/Ⅳ periodontitis (β=5.539, 95%CI: 4.442-6.636, P<0.001) had significantly higher plasma p-tau217 levels. In addition, trend test indicated that plasma p-tau217 levels increased with periodontitis severity (P_trend<0.001). After adjustments for baseline covariates, RCS regression analysis further revealed that PISA had a positive linear dose-response relationship with MCI risk (P_overall=0.002, P_nonlinear=0.344) and plasma p-tau217 levels (P_overall<0.001, P_nonlinear=0.140). After adjustments for baseline covariates, mediation analysis showed that plasma p-tau217 mediated the association between periodontitis and MCI, with a mediation proportion of 13.99% (95% Bootstrap CI: 0.38%-49.39%, P=0.038). CONCLUSIONS Periodontitis was independently positively associated with MCI risk, and plasma p-tau217 plays a mediating role in this association.
Humans ; Cognitive Dysfunction/complications* ; tau Proteins/blood* ; Periodontitis/complications* ; Case-Control Studies ; Male ; Female ; Phosphorylation ; Aged ; Middle Aged ; Activities of Daily Living

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*

Nipah virus is a highly pathogenic zoonotic virus.The viral characteristics,epidemiological features,viral life cycle and pathogenic mechanisms of Nipah virus were summarized in the article,and the alternative drugs tar-geting various stages of the viral life cycle,including antibody drugs,peptide drugs,and small molecule com-pounds,were systematically reviewed.The current challenges confronting the process of research and development of alternative drugs for Nipah virus were analyzed,and an outlook was also made on future research direction of candidate drugs for Nipah virus,aiming to provide comprehensive and in-depth theoretical reference for prevention and control of Nipah virus as well as the development of drugs and to facilitate the further development of related fields.

Objective To observe the effects of asiaticoside on renal fibrosis in diabetic nephropathy(DN)rats by regulating the Hippo-yes-associated protein(YAP)/transcriptional co-activator with PDZ-binding motif(TAZ)signaling pathway.Methods A DN rat model was established by feeding a high-sugar and high-fat diet combined with intraperitoneal injection of streptozotocin(STZ).The successfully modeled rats were randomly divided into model group,low-and high-dose asiaticoside groups,and high-dose asiaticoside+XMU-MP-1(Hippo-YAP/TAZ pathway activator)group,with a normal group set as control.After group intervention,blood urea nitrogen(BUN),serum creatinine(SCr),fasting blood glucose(FBG),and 24-hour urinary protein levels were measured.Hematoxylin-eosin(HE)staining was used to observe renal histopathological changes,Masson staining was used to assess renal fibrosis,and Western Blot was used to detect the protein expressions of YAP and TAZ in renal tissue.Results Compared with the normal group,the model group showed disordered renal tissue structure,enlarged and irregular cystic cavities,enlarged renal tubules with thickened basement membranes,inflammatory cell infiltration,and significant collagen deposition in the glomeruli,renal tubules,and vascular walls.The levels of FBG,BUN,SCr,24-hour urinary protein,and the protein expressions of YAP and TAZ in renal tissue were significantly increased(P＜0.05).Compared with the model group,the low-and high-dose asiaticoside groups showed significant improvement in renal tissue damage and fibrosis,with reduced levels of FBG,BUN,SCr,24-hour urinary protein,and decreased protein expressions of YAP and TAZ in renal tissue(P＜0.05).Compared with the high-dose asiaticoside group,the high-dose asiaticoside+XMU-MP-1 group showed a reversal in all the above indicators.Conclusion Asiaticoside may alleviate renal fibrosis and improve renal injury in DN rats by inhibiting the Hippo-YAP/TAZ pathway.

Objective To comprehensively assess the predictive value of C-reactive protein(CRP)/D-dimer(D-D)and albumin/fibrinogen(FAR)in predicting major adverse cardiovascular events(MACE)after percutaneous coronary intervention(PCI)in patients with coronary heart disease(CHD)and to construct a nomogram model for predicting post-procedural MACE in CHD patients.Methods A retrospective study was conducted on 201 CHD patients who underwent PCI at the First Affiliated Hospital of Kunming Medical University between June 2022 and March 2025.These patients were divided into MACE group(n=77)and non-MACE group(n=124)based on whether MACE occurred or not.84 CHD patients from another medical center were also collected as the validation set.The expression levels of CRP/D-D and FAR were compared between the two groups;independent predictors of postoperative MACE in CHD patients were screened by univariate and multivariate logistic regression analyses;the predictive value of CRP/D-D and FAR for the occurrence of postoperative MACE in CHD patients was assessed using ROC curves;A nomogram model was established integrating indicators such as CRP/D-D and FAR,and internal and external validations of the nomogram model were conducted using ROC curves,calibration curves,and decision curve analysis(DCA)curves.Results Compared with CHD patients in the non-MACE group,CRP/D-D and FAR levels were significantly higher in the MACE group(P<0.05).Multivariate analysis showed that age,NTproBNP,WBC,CRP/D-D,and FAR were independent risk factors for postoperative MACE in CHD patients(P<0.05).ROC curve analysis indicated that the AUC predicted by CRP/D-D combined with FAR was higher than that of CRP/D-D alone(Z=3.473,P<0.001),and FAR alone(Z=2.812,P<0.05).The Nomogram model constructed based on the aforementioned factors was validated internally and externally,and the results showed that the Nomogram model had good calibration,excellent discriminative ability,and reliable clinical utility,accurately predicting the risk of postoperative MACE.Conclusion The CRP/D-D ratio and FAR,as emerging composite biomarkers,showed significant predictive ability in predicting the risk of MACE after PCI in patients with CHD,providing a new reliable tool for clinical risk stratification.

OBJECTIVE To standardize the strategies for prevention and control of Chikungunya(CHIK)in healthcare in-stitutions so as to reduce the risk of transmission in the institutions.METHODS A working group comprising the ex-perts in hospital infection control,infectious diseases,and microbiology systematically reviewed domestic and international evidence and current guidelines,integrated China's vector ecology and healthcare realities,conducted two rounds of Delphi to achieve expert consensus,and graded the evidence and recommendation strength using the Oxford Centre for Evidence Based Medicine system.RESULTS The consensus issues 18 actionable recommendations on triage,patient mosquito-proof isolation,integrated vector control,protection of susceptible populations,environmental cleaning and disinfection,specimen management,medical textile handling,and outbreak emergency response,with each statement assigned an evi-dence level and recommendation strength.CONCLUSION This consensus is for the first time in China to provide evidence-graded strategies for control of CHIK in healthcare institutions,offering work flow-oriented,implementable guidance for clinicians,laboratorians,and infection-control personnel under different risk scenarios and enhancing the comprehensive coping capacity of the healthcare institutions.