Objective: To investigate the predictive value of EZH2 expression for malignant transformation in oral leukoplakia (OLK) and to provide a reference for clinical practice. Methods: This study was approved by the institutional ethics committee, and informed consent was obtained from all participants. A total of 114 patients diagnosed with OLK by pathological examination and treated at our hospital between November 2020 and July 2022 were initially enrolled. After excluding those with incomplete data or follow-up, 105 participants were included in the final analysis, comprising 14 in the high EZH2 expression group and 91 in the low EZH2 expression group. Histopathological examination of oral mucosa and immunohistochemical detection of EZH2 protein expression were performed. The follow-up period was 30 months; participants were followed until malignant transformation occurred or until the end of follow-up, at which point they were withdrawn from the study. The exposure factor was the level of EZH2 protein expression, and the outcome was the malignant transformation rate of OLK. Differences in EZH2 expression levels and transformation outcomes were analyzed. Results: There were no statistically significant differences between the high and low EZH2 expression groups in terms of age, sex, history of systemic disease, lifestyle habits, psychological status, diet, and sleep conditions (P > 0.05). Lesions in the high EZH2 expression group were mainly located on the ventral tongue, while in the low EZH2 expression group, they were more commonly found on the dorsal tongue and buccal mucosa. The malignant transformation rate was 28.6% (4/14) in the high expression group and 8.8% (8/91) in the low expression group; these differences were not statistically significant (P=0.053). In univariate Cox regression analysis, the risk of malignant transformation in the high EZH2 expression group was 3.647 times that of the low EZH2 expression group (HR = 3.647, 95% CI: 1.097-12.120, P<0.05). Kaplan-Meier survival analysis showed that over the 30-month follow-up period, the cancer-free survival rate in the high EZH2 expression group was 19.8% lower than in the low expression group, and the difference was statistically significant (P<0.05). In multivariate Cox regression analysis, only moderate and severe epithelial dysplasia were identified as independent risk factors for malignant transformation. The risk of malignant transformation in the moderate and severe dysplasia groups was 10.695 and 13.623 times higher, respectively, than in the mild dysplasia group (HR = 10.695, 95% CI: 2.270-50.396, P<0.05; HR=13.623, 95% CI: 1.918-96.774, P<0.05). EZH2 high expression was not an independent risk factor in the multivariate model (HR= 2.528, 95% CI: 0.752-8.500, P = 0.134). Conclusion High EZH2 protein expression is a risk factor for the malignant transformation of OLK but does not have independent predictive value.

Objective:To investigate the role of inspiratory muscle training in improving the respiratory function of pa-tients with subacute complete high cervical spinal cord injury(C2-C5).Method:From January 2020 to June 2021,60 patients with subacute complete cervical spinal cord injury(C2-C5)ASIA level A were randomly divided into experimental group(n=30)and control group(n=30).The patients in control group were conducted with routine rehabilitation training,in experimental group,inspiratory muscle training was added on the basis of routine rehabilitation treatment.Before and after training for a total of 6 weeks,maximum inspiratory pressure(Plmax),vital capacity(VC),forced expiratory vital capacity(FVC),forced expiratory volume in the first second(FEV1),maximal ventilatory volume(MVV),and dia-phragm thickness were measured,the cases of pulmonary infection were counted.Result:After the training,the improvement of PImax,VC,FVC,FEV1,MVV and diaphragm thickness in patients in experimental group were significantly better than that in patients in control group(P＜0.05),and the rate of pulmonary infection was lower(P＜0.05).Conclusion:Inspiratory muscle training added on the basis of routine rehabilitation treatment can significantly improve the respiratory function of patients with subacute complete high cervical spinal cord injury.

Objective To analyze differences in the intestinal microbiota and transcriptomics between N-methyl-N'-nitro-N-nitrosoguanidine(MNNG)gastric cancer rats and normal rats and to analyze the correlation between the two,so as to provide a reference for related studies using MNNG gastric cancer rats as a model.Methods A total of 12 Wistar rats were randomly divided into normal(NM)and gastric cancer(GC)groups.The GC group was given a concentration of 20 mg/mL of MNNG by gavage with a dose of 100 g/mL once a day,and the NM group was given the same amount of normal saline by gavage.Samples were collected for testing after 16 weeks of continuous intervention.The gastric tissues were collected and stained by HE staining to observe morphological changes in the gastric mucosa of the two groups,and the expression levels of differential genes were detected by transcriptome sequencing.The cecal contents were collected for 16S rRNA sequencing.Results(1)Visual observation and HE results showed that the volume of gastric mucosa in the NM group was normal,the surface was glossy,the gastric wall was elastic,the direction of the mucosal folds was regular,there were no hyperplasia or hemorrhagic spots.In the GC group,the volume of gastric mucosa was reduced,the gastric wall was thinned,elasticity was poor,the direction of the folds was disordered and irregular,and there was a bulge accompanied by yellow-black keratotic hyperplasia.In the NM group,the squamous epithelial layer,submucosa,and muscular layer of the gastric mucosa were clear,with no hyperplasia and keratinization.In the GC group,the gastric mucosa had disorganized layers and cell polarity,with different cell morphologies;the squamous epithelial layer was destroyed,and squamous epithelial cells were hyperplasic,keratinized,and had invaded the muscular layer by proliferation.The modeling was considered successful.(2)The results of intestinal microbiota sequencing showed that the abundance of Akkermansia and Lactobacillus in MNNG gastric cancer rats decreased significantly,and the abundance of the rumen coccaceae Prevonella,and Blauter increased significantly.(3)The three key pathways obtained by transcriptomic sequencing and KEGG pathway enrichment analysis were amebiasis,systemic lupus erythematosus,and the PI3K-Akt signaling pathway,and five genes differentially enriched in these three pathways were those for MCPT8I2,IGH-6,IGHG1,ACTN2,and VEGF-D.(4)Combined analysis of intestinal microbiota and transcriptomics showed that_UCG-005,Prevonella_UCG-003 and Brautella were positively correlated with amebiasis,systemic lupus erythematosus,and the PI3K-Akt signaling pathway.Conclusions The abundance of intestinal microbiota in gastric cancer rats formed by MNNG gavage is different from that of normal rats.The genes for MCPT8I2,IGH-6,IGHG1,ACTN2 and VEGF-D may be up-regulated in gastric cancer induced by MNNG gavage.Combined analysis of intestinal microbiota and differential genes suggested that the mechanism of MNNG carcinogenesis may be mainly related to the destruction of gastric mucosa and the inflammatory response.

In recent years,there has been a gradual increase in the number of common diseases among children in China,especially younger children.Children are a unique population with distinct differences from adults,such as indications,dosage,accessories,and other characteristics,when it comes to using medication.Long-acting injections,in comparison to regular injections used for long-term treatment in children,can reduce sudden drug side effects and significantly decrease injection frequency.This greatly improves compliance among child patients and enhances clinical outcomes.Long-acting injections offer significant advantages for managing diseases in children.This paper reviews the progress made in basic research on long-acting injectable formulations for children based on interactions between drugs and materials.Additionally,potential future development directions are discussed to guide further advancements in long-acting injections specifically designed for children's needs.

BACKGROUND:The technique of ultrasound processing is widely used for molecular biological analysis.It is of great significance to study the DNA quality of tissue with different storage years under new ultrasonic treatment for further specimen quality control of molecular detection.OBJECTIVE:To explore the effects of different storage durations on DNA quality in specimens with ultrasound processing to investigate the optimal storage time for molecular tests.METHODS:Forty specimens of breast biopsy were collected and paraffin specimens were prepared by ultrasonography.These specimens were divided into four groups based on their storage periods:<1 year,1-3 years,>3-5 years,and>5 years,which contained 10 cases in each group.Paraffin specimens were sliced;each slice was 3 μm thick;10-15 slices were taken,and DNA was extracted.The mass concentration of DNA was examined by Nanophotometer N60 ultra-micro spectrophotometer and Qubit 4.0 fluorometer.The purity of the DNA was analyzed by the ratio of A260/A280.DNA fragment integrity was measured by capillary electrophoresis (Qsep 100) to evaluate the quality of the DNA fragments.RESULTS AND CONCLUSION:The mean values of A260/A280 in the four groups were between 1.8 and 2.0,meeting the requirements of tests,without significant differences.The mean values of DNA mass concentration (Qubit concentration) were 30.39,14.33,2.52,and 1.95 ng/μL,respectively.The mean values of the N/Q were 6.48,14.18,24.56,and 29.86.The mean values of DNA were:5.64,1.76,1.24,and 0.80.The percentage of large DNA fragments averaged 56.08％,17.72％,12.68％,and 7.90％.Moreover,the Ct values of the internal control detected by PCR were 15.32,17.09,18.39,and 21.24.The three other groups exhibited significantly lower DNA concentration,higher N/Q ratios,decreased DNA quality and percentage of large fragments,and increased values of Ct,compared with the group of within 1 year of storage (P<0.05).The experimental results suggested that for novel ultrasound processed biopsy specimens,we should prioritize samples stored within 1 year for molecular testing.Samples stored within 3 years can also meet the requirements of second-generation sequencing and other tests.Samples stored within 5 years can only be attempted to carry out PCR.Samples stored for more than 5 years were not recommended to carry out molecular tests.

Objective:To explore evaluate value of chest vascular computerized tomography(CT)parameters combined with platelet-to-lymphocyte ratio(PLR)and Interleukin(IL)-17 for the prognosis of patients with acute exacerbation of chronic obstructive pulmonary disease(AECOPD)complicated with pulmonary hypertension.Methods:92 patients with AECOPD complicated with pulmonary hypertension admitted to our hospital from February 2023 to June 2024 were prospectively selected as the research objects,and they were divided into poor prognosis group(n=41)and good prognosis group(n=51)according to the different prognosis.The chest vascular CT parameters,PLR and IL-17 indicators of the two groups were compared.Multivariate Logistic regression analysis were used to analyze the influencing factors of prognosis in patients with AECOPD complicated with pulmonary hypertension.The receiver operating characteristic(ROC)curve was used to analyze the evaluate value of chest vascular CT parameters,PLR and IL-17 for the prognosis of patients with AECOPD complicated with pulmonary hypertension.Results:Among 92 patients with AECOPD complicated with pulmonary hypertension,41 patients were readmitted for treatment,and the incidence of poor prognosis was 44.57％.The age and disease duration of patients in poor prognosis group were higher than those in good prognosis group(P＜0.05).The ascending aortic diameter(AAD),main pulmonary artery diameter(MPAD),descending aortic diameter(DAD),PLR and IL-17 in poor prognosis group were higher than those in the good prognosis group(P＜0.05).Multivariate logistic analysis showed that elevated MPAD,AAD,DAD,IL-17,PLR,advanced age and long disease course were risk factors for poor prognosis in patients with AECOPD complicated with pulmonary hypertension.ROC curve analysis showed that the area under the curve(AUC)of the chest vascular CT parameters MPAD,AAD,DAD combined with PLR and IL-17 for evaluating the prognosis of AECOPD complicated with pulmonary hypertension was 0.923,which was significantly higher than that evaluated by each index alone.Conclusion:Elevated MPAD,AAD,DAD,IL-17,PLR,advanced age and long disease course are risk factors for poor prognosis in patients with AECOPD complicated with pulmonary hypertension,the combined detection of chest vascular CT parameters and PLR,IL-17 has a high evaluating value for the prognosis of patients with AECOPD complicated with pulmonary hypertension.

Objective To explore function-structure covariant patterns in Alzheimer's disease(AD)and mild cognitive impairment(MCI),and to investigate their associations with cognitive function and activities of daily living.Methods three-way parallel group independent component analysis(three-way pGICA),was used to identify the covariant patterns of resting-state functional MRI temporal data,gray matter density maps,and fractional anisotropy(FA)maps,and the differences between different groups were compared.Furthermore,the associations of covariant patterns with the Montreal Cognitive Assessment-Basic(MoCA_B)Scale scores and Activities of Daily Living Scale scores were analyzed.Results The function-structure covariant patterns in AD and MCI were characterized by the enhanced negative functional connectivity between the left posterior salience network and the right default mode network,the decreased gray matter density in the bilateral dorsolateral prefrontal cortex,and the reduced FA values in the left superior corona radiata(correlations:P<0.001,FDR corrected).Compared with HC group,AD group showed significant abnormalities in all identified covariant patterns(P<0.01,FDR corrected),but MCI group only exhibited a significant decrease in gray matter density in the bilateral dorsolateral prefrontal cortex(P<0.05,FDR corrected).Additionally,AD group had significantly lower FA value in the left superior corona radiata than MCI group(P<0.05,FDR corrected).The loadings reflecting the degree of covariation were significantly correlated with the Activities of Daily Living Scale scores(P<0.05,FDR corrected)but not with MoCA_B Scale scores.Conclusion The function-structure covariant patterns in AD and MCI are consistent with the declines in activities of daily living.The multimodal fusion analysis(three-way pGICA)provides a novel approach to understand the brain damage mechanisms underlying the covariant evolution of MCI and AD.

Objective To conduct a comprehensive review on application status and future development prospects of proton therapy for pediatric medulloblastoma.Methods A total of 218 literatures were retrieved from PubMed and CNKI database using the search terms"pediatric medulloblastoma,proton therapy,radiotherapy"(English)and"儿童髓母细胞瘤,质子治疗,放射治疗"(Chinese),with a publication timeframe from January 1,2004,to June 1,2025.Inclusion criteria were as follow:(1)proton therapy for pediatric medulloblastoma;(2)radiotherapy for pediatric medulloblastoma;(3)proton therapy for pediatric brain tumors;(4)development and applications of proton therapy.Exclusion criteria were as follow:(1)outdated literatures;(2)redundant or highly similar studies.After screening,89 literatures met the inclusion criteria.Results Compared with conventional treatments such as surgery,photon therapy,and chemotherapy,proton therapy for pediatric medulloblastoma significantly reduced acute toxicity and long-term side effects including cognitive dysfunction,endocrine disorders,and hearing loss.Additionally,proton therapy exhibited favorable cost-effectiveness.In the future,the therapeutic outcomes would be further enhanced through the optimization of proton therapy techniques,treatment planning,and equipment.Conclusion With ongoing technological advancements and growing clinical experience,proton therapy is expected to become one of the standard treatment modalities for pediatric medulloblastoma.

Objective To establish a rapid on-site detection method for methamphetamine(MA)based on surface-enhanced Raman spectroscopy(SERS).Methods Laboratory-synthesized colloidal silver nanoparticles(AgNPs)were employed as SERS substrates to rapidly detect methamphetamine in typical crime scene matrices,including beer,cigarette ash,and plastic containers.Results The method requires no sample pretreatment,is simple to operate,and offers high sensitivity with fast detection speed.The limit of detection(LOD)reached 10 ppb(signal-to-noise ratio,S/N＞3).The qualitative results of methamphetamine analysis from forensic case samples were highly consistent with gas chromatography-mass spectrometry(GC-MS)findings.Conclusion Methamphetamine is frequently encountered in illicit settings and criminal activities.This study achieved rapid detection of methamphetamine in complex matrices(beer,cigarette ash,and plastic containers),demonstrating good application prospects in practice and providing important value for law enforcement,public safety,and criminal investigation.

DNA methylation is an important epigenetic modification in mammals, playing a crucial role in various physiological processes, including cell differentiation and the gene expression regulation. The ten-eleven translocation (TET) protein family of DNA demethylases is integral to the regulation of DNA methylation, as it catalyzes the oxidation of 5-methylcytosine to form 5-hydroxymethylcytosine. During early embryonic development, the genome undergoes extensive DNA demethylation, and any aberration in this reprogramming process can result in abnormal embryonic development and physiological defects in offspring. The TET proteins, due to their unique dynamics and multifaceted roles, facilitate DNA demethylation and are involved in development and maturation of germ cells, the establishment of pluripotency, cell lineage differentiation, and transcriptional processes throughout mammalian embryogenesis. Furthermore, these proteins are closely associated with the maintenance of pregnancy and susceptibility of progeny to disease. Factors such as genetic mutations, maternal health conditions, and exposure to adverse environmental influences can impact TET protein activity, resulting in abnormal patterns of DNA demethylation. A comprehensive investigation of the related mechanisms of TET proteins is essential for enhancing our understanding of epigenetic regulation during early life, diagnosing and treating related diseases such as early fetal development retardation, and informing strategies for the prevention and management of pregnancy.This article reviews the regulatory role of DNA demethylation mediated by TET protein in mammalian embryonic development and pregnancy outcomes.