Objective: To investigate the efficacy of magnesium-strontium co-doped hydroxyapatite mineralized collagen (MSHA/Col) in improving the bone repair microenvironment and enhancing bone regeneration capacity, providing a strategy to address the insufficient biomimetic composition and limited bioactivity of traditional hydroxyapatite mineralized collagen (HA/Col) scaffolds. Methods: A high-molecular-weight polyacrylic acid-stabilized amorphous calcium magnesium strontium phosphate precursor (HPAA/ACMSP) was prepared. Its morphology and elemental distribution were characterized by high-resolution transmission electron microscopy (TEM) and energy-dispersive spectroscopy. Recombinant collagen sponge blocks were immersed in the HPAA/ACMSP mineralization solution. Magnesium-strontium co-doped hydroxyapatite was induced to deposit within collagen fibers (experimental group: MSHA/Col; control group: HA/Col). The morphological characteristics of MSHA/Col were observed using scanning electron microscopy (SEM). Its crystal structure and chemical composition were analyzed by X-ray diffraction and Fourier transform infrared spectroscopy, respectively. The mineral phase content was evaluated by thermogravimetric analysis. The scaffold's porosity, ion release, and in vitro degradation performance were also determined. For cytological experiments, CCK-8 assay, live/dead cell staining, alkaline phosphatase staining, alizarin red S staining, RT-qPCR, and western blotting were used to evaluate the effects of the MSHA/Col scaffold on the proliferation, viability, early osteogenic differentiation activity, late mineralization capacity, and gene and protein expression levels of key osteogenic markers [runt-related transcription factor 2 (Runx2), collagen type Ⅰ (Col-Ⅰ), osteopontin (Opn), and osteocalcin (Ocn)] in mouse embryonic osteoblast precursor cells (MC3T3-E1). Results: HPAA/ACMSP appeared as amorphous spherical nanoparticles under TEM, with energy spectrum analysis showing uniform distribution of carbon, oxygen, calcium, phosphorus, magnesium, and strontium elements. SEM results of MSHA/Col indicated successful complete intrafibrillar mineralization. Elemental analysis showed the mass fractions of magnesium and strontium were 0.72% (matching the magnesium content in natural bone) and 2.89%, respectively. X-ray diffraction revealed characteristic peaks of hydroxyapatite crystals (25.86°, 31°-34°). Infrared spectroscopy results showed characteristic absorption peaks for both collagen and hydroxyapatite. Thermogravimetric analysis indicated a mineral phase content of 78.29% in the material. The scaffold porosity was 91.6% ± 1.1%, close to the level of natural bone tissue. Ion release curves demonstrated sustained release behavior for both magnesium and strontium ions. The in vitro degradation rate matched the ingrowth rate of new bone tissue. Cytological experiments showed that MSHA/Col significantly promoted MC3T3-E1 cell proliferation (130% increase in activity at 72 h, P < 0.001). MSHA/Col exhibited excellent efficacy in promoting osteogenic differentiation, significantly upregulating the expression of osteogenesis-related genes and proteins (Runx2, Col-Ⅰ, Opn, Ocn) (P < 0.01). Conclusion The MSHA/Col scaffold achieves dual biomimicry of natural bone in both composition and structure, and effectively promotes osteogenic differentiation at the genetic and protein levels, breaking through the functional limitations of pure hydroxyapatite mineralized collagen. This provides a new strategy for the development of functional bone repair materials

Background: Advances in intensive care have markedly improved survival after severe brain injury, leading to a growing population of patients with prolonged disorders of consciousness (pDOC). Current management of pDOC remains largely supportive, and evidence-based neuromodulatory interventions are limited; moreover, existing guidelines provide insufficiently explicit recommendations regarding mechanisms of action and objective biomarkers of treatment response. Transcutaneous auricular vagus nerve stimulation (taVNS) has emerged as a potential noninvasive intervention; however, its modulatory effects on brain function in pDOC are not yet well characterized, and the paucity of integrative mechanistic evidence has constrained its translation into routine clinical practice. Objectives: Within a multimodal assessment framework, this study aims to systematically elucidate the neurobiological mechanisms by which taVNS modulates brain function and autonomic activity in patients with pDOC, and to evaluate its clinical potential to enhance levels of consciousness. Methods: In this randomized controlled trial, 60 patients with vegetative state/minimally conscious state will be enrolled and randomly allocated to a taVNS group, a transcutaneous nonauricular vagus nerve stimulation group (sham), or a control group (n = 20 per group) for a 4-week intervention. The primary outcome will be changes in the Coma Recovery Scale-Revised scores from baseline to weeks 1, 2, and 4 of treatment. Secondary outcomes will include functional brain activity assessed by electroencephalography and functional near-infrared spectroscopy, as well as autonomic modulation indexed by heart rate variability. Functional prognosis will be evaluated using the Glasgow Outcome Scale-Extended at the end of treatment and at a 6-month follow-up. Safety will be assessed by continuous monitoring and documentation of adverse events throughout the study period. Results and discussion: By integrating electroencephalography–functional near-infrared spectroscopy with heart rate variability, this study will characterize the effects of taVNS on functional brain networks and consciousness recovery in pDOC across complementary behavioral, electrophysiological, hemodynamic, and autonomic domains, while interrogating potential sources of clinical and neurobiological heterogeneity. The findings are expected to provide a mechanistic and evidence-based foundation for the mechanism-driven clinical implementation of taVNS and the optimization of stimulation protocols in pDOC. Clinical trial registration: International Traditional Medicine Clinical Trial Registry, ITMCTR20250021041, https://itmctr.ccebtcm.org.cn.

Factorial design is an important method for optimizing complex strategies in implementation science. This paper systematically elucidates the origin, types, and core concepts of the factorial design, and then use a brief smoking cessation intervention project as an example to illustrate its specific applications in identifying key factors and testing interactions. Factorial design has the following typical advantages: it can test synergistic and antagonistic effects between factors, provide effect estimates that are broadly valid across the experimental conditions, and offer high statistical efficiency. At the same time, it also has certain limitations, including that its complexity is positively correlated with the number of factors, the need for large sample sizes to detect interactions, the potential for insufficient acceptability of some strategy combinations due to ethical concerns or participant burden, and the complexity of the analytical methods required. In future research, its application should be further optimized by appropriately controlling the number of factors, clearly defining research objectives, and evaluating intervention acceptability, in order to promote the development of precision implementation science.

Objective: To evaluate oral health status and knowledge, attitude and practice (KAP) among third grade primary school students after the implementation of the Comprehensive Intervention Project for Childhood Oral Diseases in Guizhou Province, so as to provide evidence for program optimization. Methods: Using a multistage cluster sampling method, oral examinations and questionnaire surveys were conducted among third grade primary school students in schools covered by the Comprehensive Intervention Project for Childhood Oral Diseases in Guizhou Province (intervention group, n =1 030) and schools not covered by the program (control group， n =1 130) in November 2023. A follow up survey was conducted one year after the project s implementation (December 2024) to evaluate the differences in oral health status and KAP between the two groups. The t-test and χ 2 test were used to compare means and rates, respectively. Results: One year after implementation, the permanent caries prevalence, caries filling ratio, mean dmft/DMFT, and mean simplified debris index in the intervention group were 33.3%, 22.3 %, 3.44±3.10, 0.77±0.56, which were all better than those in the control group (39.8%, 17.3%, 3.89±2.85, and 0.87± 0.63 ) ( χ 2/ t =9.87, 8.78, -2.59, -3.91,all P <0.05). After intervention, the levels of oral health knowledge, attitude and behaviors in the intervention group were better than those in the control group ( Z =7.69, 2.35, 2.10, all P <0.05). Except for the item "fluoride has no protective effect on teeth", the correct response rates of other oral health knowledge items in the intervention group were higher than those in the control group ( χ 2=3.92-163.94, all P <0.05). The reported rates of tooth brushing related behaviors and sugary beverage consumption were higher in the intervention group than in the control group ( χ 2=4.02-11.42，all P <0.05). Conclusions The Comprehensive Intervention Project for Childhood Oral Diseases improves children s oral health status, oral health knowledge, and some oral health behaviors, but has limited effects on some behaviors and oral health attitudes. Future efforts should focus on the factors influencing the transformation of children s oral health attitudes and behaviors, along with strengthening long term follow up evaluations.

Vancomycin is a glycopeptide antibiotic widely used for severe infections caused by gram-positive bacteria. Nevertheless， its clinical application is limited by a narrow therapeutic window and inter-individual differences in pharmacokinetics. Conventional dosing regimens rely on clinical experience and population pharmacokinetic models， which make precise therapy difficult. The emergence of machine learning provides new insights into tackling the above challenges. By integrating multi-dimensional data including demographic characteristics， clinical indicators and combined medications， machine learning can assist individualized medication for patients and improve the capacity for vancomycin dose prediction， in vivo exposure assessment and renal toxicity risk warning. However， the application of machine learning still faces multiple challenges such as poor data quality， weak model generalization ability and insufficient interpretability. Its clinical value needs to be further confirmed by prospective studies and external validation. Future research may consider deep integration of machine learning and population pharmacokinetic models to better support vancomycin individualized medication.

This paper summarizes the experience of Professor TU Jinwen in the staged treatment of oculomotor nerve palsy （ONP） based on the theory of qi collaterals. In the acute onset stage， the key pathogenesis is triggered by wind toxin and disharmony of the qi collaterlas， for which the treatment should prioritize the elimination of pathogenic factors， with therapeutic principles of dispelling wind and eliminating toxin， boosting qi and harmonizing collaterals， using modified Qianzheng Powder （牵正散） plus Buzhong Yiqi Decoction （补中益气汤）. In the remission and recovery stage， the key pathogenesis is stagnation of blood stasis and damage to the qi collaterals， it is recommended to treat the branch， with principles of boosting qi and activating blood， resolving stasis and unblocking collaterals， using the modified Buyang Huanwu Decoction （补阳还五汤）. In the sequela and protracted stage， the key pathogenesis is depletion of the liver， spleen， and kidney with non-nourishment of the qi collaterals； the treatment should focus on addressing the root cause， with principles of fortifying the spleen and boosting qi， tonifying the liver and kidney， using the modified Sijunzi Decoction （四君子汤） combined with Qiju Dihuang Pill （杞菊地黄丸）.

BACKGROUND:Traditional methods of urinary tract reconstruction are limited by donor scarcity,high complication rates,and suboptimal functional recovery.Tissue engineering strategies offer new directions in this field.Since the urinary tract is mainly composed of muscle tissue,the key is to find suitable seed cells and efficiently induce them to differentiate into smooth muscle cells.Comparative studies on the efficacy of different smooth muscle cell induction regimens are still lacking. OBJECTIVE:To isolate,culture,and identify human urine-derived stem cells,and to compare the effects of two different induction protocols. METHODS:Human urine-derived stem cells were isolated from urine samples of 11 healthy adult volunteers by multiple centrifugations.Surface markers were identified by flow cytometry.The multi-directional differentiation potential of human urine-derived stem cells was verified through osteogenic and adipogenic differentiation.Differentiation was induced by transforming growth factor-β1 or transforming growth factor-β1 combined with platelet derived growth factor for 14 days.Immunofluorescence staining and western blot assay were employed to compare the expression differences of smooth muscle-specific proteins(α-SMA and SM22). RESULTS AND CONCLUSION:(1)Urine-derived stem cells were successfully isolated from the eight urine samples of healthy people.These cells exhibit a"rice grain"-like morphology and possess a robust proliferative capacity.(2)Urine-derived stem cells exhibited high expression of mesenchymal stem cell surface markers(CD73,CD90,and CD44)and extremely low expression of hematopoietic stem cell surface markers(CD34 and CD45).These cells did not express CD19,CD105,and HLA-DR.(3)After osteogenic and adipogenic differentiation,the formation of calcium nodules and lipid droplets was observed,with positive staining results from Alizarin Red S and Oil Red O staining.(4)After 14 days of smooth muscle induction culture,immunofluorescence staining revealed that the smooth muscle differentiation rate of urine-derived stem cells treated with a combination of transforming growth factor-β1 and platelet derived growth factor was significantly higher compared to those treated with transforming growth factor-β1 alone(P<0.005).(5)After 14 days of smooth muscle induction culture,western blot assay further demonstrated that the expression levels of α-SMA and SM22 in the transforming growth factor-β1/platelet derived growth factor group were significantly elevated compared to those in the transforming growth factor-β1 only group(P<0.005).These findings confirm that urine-derived stem cells can be non-invasively isolated using multiple rounds of centrifugation.Compared with transforming growth factor-β1 alone,the combination of transforming growth factor-β1 and platelet derived growth factor can improve the efficiency of inducing urine-derived stem cells to differentiate into smooth muscle cells.

To summarise the clinical experience of treating perimenopausal syndrome with self-prescribed Gengnian Formula （更年方）. It is believed that the key mechanism of perimenopausal syndrome is the imbalance of heart and liver， and it is common to see the syndrome of blazing of heart-liver fire， so the treatment should be based on calmness of qi and blood. It is advocated that using the method of clearing the heart and pacifying liver， prescribing Gengnian Formula as the basic formula， and modifying according to the symptoms， so as to regulate heart and liver and balance yin and yang.

BACKGROUND:Bile acid metabolism plays a crucial role in the development and progression of Crohn's disease.There is no research on changes in bile acid metabolism and key target genes following treatment with biological agents.OBJECTIVE:To investigate the expression characteristics of bile acid metabolism-related genes in patients with Crohn's disease,identify key genes associated with response to biological agents.METHODS:Transcriptome data were obtained through the GEO database to analyze differentially expressed genes between inflammation-control groups and inflammation-treatment groups.GO,KEGG,and GSEA enrichment analyses were used to evaluate the effects of biological agent therapy on bile acid metabolism.Protein-protein interaction network and WGCNA algorithm were employed to analyze differentially expressed genes,identifying modules closely related to biological agent treatment response,which led to the determination of UGT2A3 as a key gene in bile acid metabolism.In the inflammation group of the GSE186582 dataset,samples were divided into high and low expression groups based on UGT2A3 levels to study its relationship with immune infiltration and explore the interaction between UGT2A3 and the immune microenvironment.Clinical characteristics and intestinal manifestations were compared between high and low expression groups,and correlations between UGT2A3 and clinical indicators(C-reactive protein,erythrocyte sedimentation rate,Crohn's disease activity index,and Crohn's disease endoscopic activity score)were investigated.The competing endogenous RNA regulatory network of UGT2A3 was constructed,and its upstream miRNA was functionally enriched to explore the molecular mechanism of UGT2A3 in bile acid metabolism.Single-cell analysis and clustering were performed using high-throughput sequencing data of GSE134809 to observe the expression of UGT2A3 in different samples and cell populations.Colon tissue samples from untreated and biologic-treated Crohn's disease patients and healthy colon tissue samples from patients with intestinal polyps were collected,and UGT2A3 expression was detected by immunohistochemistry,qRT-PCR,and western blot assay.Fresh feces from Crohn's disease patients and healthy controls were collected to detect bile acid levels,and the relationship between UGT2A3 and fecal bile acid levels was analyzed.RESULTS AND CONCLUSION:A total of 11 bile acid metabolism-related genes were screened,showing significant changes in gene expression after biological agent therapy.GO and KEGG enrichment analyses revealed that intestinal nutrient absorption and metabolic processes normalized after treatment,while leukocyte chemotaxis and inflammatory response pathway activity decreased.GSEA analysis revealed significant enrichment of bile acid metabolism-related pathways after treatment.Protein-protein interaction network construction and WGCNA analysis identified UGT2A3 as a key gene closely associated with treatment response.UGT2A3 expression was significantly decreased in inflamed tissues of Crohn's disease patients and returned to normal levels after biological agent therapy.This result was confirmed in clinical specimens.UGT2A3 expression levels showed significant negative correlations with C-reactive protein,erythrocyte sedimentation rate,Crohn's Disease Activity Index,and Crohn's Disease Endoscopic Index of Severity.Receiver Operating Characteristic curve analysis demonstrated that UGT2A3 has good diagnostic value(Area Under Curve AUC=0.801 0)and effectively reflects treatment outcomes.Immune infiltration analysis showed significantly increased infiltration of various immune cells in samples with low UGT2A3 expression,and its expression levels negatively correlated with immune scores,microenvironment scores,and stromal scores.Compared with the low UGT2A3 expression group,patients with high expression showed less fecal occult blood and penetrating inflammation,with milder intestinal strictures and general condition severity.Fecal bile acid analysis revealed that UGT2A3 expression strongly negatively correlated with primary bile acid content and strongly positively correlated with secondary bile acid content.

Environmental pollutants generated by industrialization,characterized by microplastics,are increasingly impairing male fertility in the form of"environmental hidden toxins".Although the concept of environmental toxicity has been proposed for a long time,its practical application remains limited.Therefore,this study innovatively introduces and elaborates on the concept of"environmental hidden toxins",highlighting its pathogenic characteristics,including latency,turbidity,consumptive nature,concurrent nature,and transmutation.Environmental hidden toxin can be classified into five categories based on the properties:stagnation toxins,scorching toxins,turbid toxins,desiccating toxins,and latent toxins.The core pathogenesis of environmental hidden toxins-induced male infertility involves three stages:firstly,invading the lungs and spleen,leading to the gradual depletion of healthy qi;secondly,forming toxin-stasis complexes that obstruct the spermatic pathway;and lastly,penetrating deeply into the seminal chamber,directly damaging the genuine essence.The treatment principle emphasizes"strengthening the foundation and clearing the source,"advocating dual strategies of detoxification and strengthening vital qi.This includes enhancing clarity and lowering turbidity,fortifying the earth to generate metal to replenish healthy qi;resolving stasis,unblocking collaterals,and dispelling toxins to eliminate pathogenic factors;and nourishing the kidneys,replenishing essence,and expelling toxins to preserve the genuine essence.All the above approaches form a holistic traditional Chinese medicine(TCM)strategy that treating both manifestation and root cause of disease.The study provides theoretical foundations and offers novel clinical insights into TCM interventions for male infertility caused by environmental pollutants.