The repair of bone defects is heavily influenced by the dynamic osteogenic microenvironment. Static scaffolds constructed by traditional 3D printing technology cannot simulate the dynamic nature of the microenvironment during bone defect repair due to the fixed structure, uncontrollable release of active factors, and difficult regeneration of blood vessels, among other factors. Breaking through the limitations of these static scaffolds and realizing the intelligent and dynamic regulation of the osteogenic microenvironment is a key scientific issue in the field of bone tissue engineering. 4D printing technology combines the dynamic responsiveness of bone restoration materials with the concept of intelligent design to regulate the micro and macro structure of scaffolds. This technology provides a new method for bone tissue engineering by responding to endogenous and exogenous stimuli and creating a better osteogenic microenvironment through functionalized design, including drug delivery and antibacterial function. However, this technology currently suffers from challenges related to dynamic response material design, insufficient precision of printing technology, and mismatches between multi-stimulus response systems, metabolic rhythms of bone tissue, and functionalized composite scaffolds. Future research should focus on the development of smart response materials with excellent dynamic responses and bioactivity, the creation of new printing technologies, and the design of personalized and precise bone repair solutions. The aim of this paper is to review the current research status of 4D printing for bone tissue engineering in terms of material types, response mechanisms, and applications to provide a theoretical basis for the development and clinical application of functional bone repair materials in the future.

[摘 要] 目的：探讨乙醛脱氢酶1A1（ALDH1A1）mRNA负载树突状细胞（DC）疫苗体外诱导抗鼻咽癌干细胞免疫反应的能力。方法：体外转录ALDH1A1 mRNA并电转染至人外周血DC，构建ALDH1A1-DC疫苗，转染EGFP mRNA为对照。流式细胞术检测DC表型、淋巴细胞亚群、细胞毒性T 淋巴细胞（CTL）增殖和细胞因子分泌。分选ALDH+和ALDH- C666-1细胞，检测干细胞标志物（CD24、CD133）和干性基因（OCT4、SOX2、Nanog）的表达水平，采用CCK-8法评估ALDH1A1-DC疫苗诱导的特异性免疫杀伤能力。结果：EGFP-DC荧光表达率显著高于未转染DC（P < 0.000 1），ALDH1A1-DC抗原提呈分子HLA-ABC、共刺激分子CD86、CD40和CCR7表达与未转染DC相比显著增强（P = 0.001 3、P = 0.005 9、P = 0.000 4及P = 0.001 9）。与单独淋巴细胞组（T）及空载DC诱导的淋巴细胞组（DC-T）相比，ALDH1A1-DC诱导的CTL比例最高（均P < 0.000 1），分裂指数最高（P = 0.000 2、P = 0.000 3），且CD8+CD69+（P = 0.000 5、P < 0.000 1）和CD8+ IFN-γ+（P = 0.012 6、P = 0.001 9）细胞亚群比例均显著提高。ALDH1A1-DC-T细胞对ALDH+C666-1悬浮细胞球的杀伤效果显著优于ALDH-细胞球（P = 0.001 0），且与其他各组淋巴细胞相比表现出较强的抗ALDH+C666-1悬浮细胞球的免疫效应（P = 0.001 0，P < 0.000 1）。然而，HLA-ABC阻断后其对ALDH+C666-1细胞的特异性杀伤效应显著降低（P < 0.000 1）。结论：mRNA转染DC后可翻译蛋白并促进DC成熟，增强其诱导特异性CTL免疫反应的能力。

Objective:To investigate the role of the structural gene vgrG of the type Ⅵ secretion system (T6SS) of Klebsiella pneumoniae ( Kpn), and evaluate the growth ability in vitro and pathogenicity of the bacteria after vgrG was deleted. Methods:Using sequences published by the National Center for Biotechnology Information (NCBI), primers were designed to amplify the upstream and downstream homology arms of vgrG via PCR. These fragments were cloned into the vector pKO3-Km after overlapping, the recombinant vector pKO3-Km- vgrG was transferred into Kpn competent cells, and the vgrG deletion strain Δ vgrG was obtained through homologous recombination. The vgrG promoter with the complete gene fragment was amplify by PCR and cloned into the pBAD33 vector. The pBAD33- vgrG was then transferred into Δ vgrG competent cells to obtain the complemented strain CΔ vgrG. The wild-type strain (WT), Δ vgrG strain and CΔ vgrG strain were cultured in LB (Luria-Bertani) liquid medium to compare growth rates. Adhesion to human lung epithelial A549 cells and intracellular survival in macrophages Raw264.7 cells were assessed. In vivo experiments included mouse survival analysis ( n=10) and lung bacterial load quantification ( n=6). Statistical comparisons were performed using the Student t-test. Results:The Δ vgrG strain was obtained through homologous recombination. It was identified by specific primers that compared with the WT strain, the complete vgrG fragment (2 487 bp) was deleted. On this basis, the CΔ vgrG strain was obtained. Deletion of vgrG did not significantly affect Kpn growth in vitro growth ability of bacteria before on after Kpn deleted vgrG [(1.40±0.10) vs (1.20±0.30), t=0.63, P>0.05]. The viscosity of WT strain was significantly higher than that of the Δ vgrG strain [(0.96±0.04) vs (0.38±0.05), t=9.72, P<0.05], the viscosity of the CΔ vgrG strain was also significantly higher than that of the Δ vgrG strain ( P<0.05). At the cellular level, the amount of adherent bacteria of the WT strain to A549 cells was significantly greater than that of the Δ vgrG strain [(5 367.00±318.00) CFU vs (4 067.00±88.19) CFU, t=3.94, P<0.05], the amount of adherent bacteria of CΔ vgrG strain was also significantly higher than that of Δ vgrG strain ( P<0.05). After 12 h infection, the WT strain survival rate in macrophages was significantly higher than that of the Δ vgrG strain[(69.00±1.00)% vs (47.50±2.50)%, t=7.99, P<0.05]. At the animal level, the survival rate of WT strain group after lethal dose infection of mice was significantly lower than that of Δ vgrG strain group [(16.67±8.82)% vs (53.33±6.67)%, t=3.32, P<0.05]; mice infected with semi-lethal dose and the number of bacteria load in the lungs of WT strain group was significantly higher than that of the Δ vgrG strain group[ (4.97±0.06) lg CFU/g vs (4.05 ±0.04) lg CFU/g, t=12.27, P<0.01], the amount of bacteria in the lungs of mice in CΔ vgrG strain group was also significantly higher than that in Δ vgrG strain group ( P<0.01). Conclusions:The vgrG gene does not affect the growth of Kpn in vitro, but it is involved in the adhesion of Kpn to epithelial cells, resistance to macrophage killing and pathogenicity to mice.

Objective To observe the value of dynamic attention aggregation network(DAANet)for automatic segmentation of anomalous origin of the left coronary artery from the pulmonary artery(ALCAPA)in CT angiography(CTA).Methods CTA data in 30 patients with ALCAPA syndrome were retrospectively enrolled.ALCAPA were segmented in CTA with DAANet based on residual edge feature(REF),pyramid dynamic attention(PDA)and dynamic global feature aggregation(DGFA)modules.Taken physician manual segmentation results as references,the results were compared with those of other networks.Results Compared with other networks,DAANet had better value for automatic segmentation of ALCAPA in CTA,which could show the details of irregular margins of left coronary artery with good continuity and high clarity,and the performance parameters were overall better.Conclusion DAANet had better performance for automatic segmentation of ALCAPA in CTA.

This article mainly introduces Professor Xu Zhenye's experience in treating advanced colorectal cancer.According to Professor Xu,the pathogenesis of advanced colorectal cancer is mainly based on deficiency,supplemented by evils and realities,and most of the patients are suffering from deficiency of essence and Qi.The treatment should focus on replenishing the deficiency,followed by removing the evils and poisons.Professor Xu innovatively put forward the theory of"spleen deficiency and essence deficiency,supporting the positive to fight against cancer",treating intestinal cancer by strengthening the spleen,tonifying the kidney and nourishing the essence,supplemented by clearing heat and removing toxins to fight against tumour.

Objective This study investigated feasible cases and their significance in promoting the"generalized"development of inspection through digital-intelligent equipment.Methods A qualitative research approach was used,involving interviews conducted between February 2025 and March 2025 with experts in traditional Chinese medicine diagnostics,clinical research methodology,medical engineering integration,and related disciplines,using both online and offline methods.In accordance with the Consolidated Criteria for Reporting Qualitative Research,feasible cases involving the specific application of digital equipment in various parts of observation were collected through item enrichment.The significance of extending observation capabilities via these cases was analyzed,along with the overall implications of integrating digital technologies with traditional inspection method.Results Interviews were completed with 11 experts from domestic universities and research institutes in the fields of traditional Chinese medicine diagnosis,medical engineering integration,and related disciplines.A total of 78 feasible cases of digital-intelligent inspection were identified,along with 69 insights regarding the significance of enhancing the inspection capabilities.These insights were synthesized into two dimensions and 23 holistic meanings.The first dimension is to expand the scope of inspection,including obtaining internal environmental characteristics,observing external environmental characteristics,expanding thermodynamic characteristic data,and crossing time and space.The second dimension is to improve the quality of observation and diagnosis information collection and analysis,including 19 specific meanings,such as standardized collection environment,objective quantification,and refined observation.Conclusion Digital-intelligent equipment plays a significant role in expanding the scope of inspection content and achieving high-quality acquisition and analysis of extensive inspection information.These advancements extend and enrich the capabilities of traditional inspection method in traditional Chinese medicine.

Objective:To explore the effect of neck muscle group exercise on setup errors in postoperative radiotherapy for head and neck cancers.Methods:A total of 126 head and neck cancer patients scheduled for radiotherapy at the First Affiliated Hospital, Air Force Medical University from February 2021 to October 2022 were prospectively enrolled. Among these patients, four patients discontinued treatment due to personal reasons, and the remaining 126 patients were randomly divided into an experimental group and a control group, with 61 patients in each group. The experimental group received neck muscle group exercise, while the control group received routine treatment without intervention. Cone-beam CT (CBCT) scans were performed weekly to measure setup errors at the levels of the clivus and the 4 th and 7 th cervical vertebrae (C4 and C7, respectively). Three-dimensional displacement, systematic errors, and random errors were calculated for each level. The appropriate margins of planning target volumes (PTVs) were determined using the Van Herk formula. Results:Baseline characteristics were well-balanced between the two groups, with no statistically significant differences ( P > 0.05). Compared to the control group, the experimental group showed significantly smaller setup errors in the left-right ( x) and anterior-posterior ( z) directions at the clivus level, as well as in the z direction at the C4 and C7 vertebral levels ( t = 2.30, 5.29, 4.07, 2.40, P < 0.05). The required PTV margin in the z direction increased to 4.0 mm at C7 from 2.4 mm and 2.8 mm at the clivus in the experimental and control groups, respectively. Correlation analysis revealed a strong negative correlation between the x-direction at the clivus and C4 vertebral levels and the couch angle (RTN) among all patients ( r = -0.548, -0.452, P < 0.001). A moderate negative correlation was observed between the inferior-superior ( y) direction and the z-direction at the C4 and C7 vertebral levels ( r = -0.160, -0.222, P < 0.001). Conclusions:Neck muscle group exercise can reduce setup errors and PTV margins in the anterior-posterior direction in postoperative radiotherapy for head and neck cancer patients.

As one of the core theories of spleen governing transportation and transformation in the traditional Chinese medicine visceral manifestation theory,the modern biological basis of"spleen qi disperses essence"has not been fully elucidated.Lipids are one of the three major nutrients in the body,which are derived from exogenous absorption or endogenous transformation,and belong to the category of"grease"and"essence"substances in traditional Chinese medicine.Because of their hydrophobic nature,lipids require apolipoproteins to be transported in the bloodstream and used by the body;similarly,essence also needs spleen qi transformation to be distributed throughout the body and exert their nourishing effects,revealing a certain degree of inherent unity between the two.When the spleen qi functions properly,essence dispersal is orderly and lipid metabolism remains in homeostatic balance;if spleen deficient leads to impaired transportation,the essence will not be distributed,and the lipid turbidity will accumulate,causing disease.Classic strengthening spleen prescriptions such as Zexie Decoction,can reshape lipid homeostasis by regulating apolipoproteins.Based on apolipoprotein-mediated lipid metabolism,this paper explores the modern molecular biology basis of the theory of"spleen qi disperses essence,"which provides novel insights for enriching the modern research of traditional Chinese medicine visceral manifestation theory,and lays the foundation for clinical practice and theoretical innovation in the treatment of metabolic diseases from the spleen.

BACKGROUND:Spinal cord injury is a serious neurological disease that often results in severe neurological impairment. Oxidative stress is an important link in the pathological process after spinal cord injury,leading to neuronal cell death and loss of function. In recent years,natural products have shown potential application values in the treatment of oxidative stress after spinal cord injury due to their wide range of sources,diverse structures and rich biological activities. OBJECTIVE:To discuss the therapeutic effects of some natural products on oxidative stress after spinal cord injury and the related mechanisms,in order to provide new ideas and directions for antioxidant therapy of spinal cord injury. METHODS:"Spinal cord injury,oxidative stress,anti-oxidation,natural products,natural compounds,polyphenols" were used as the English and Chinese keywords in PubMed,Web of Science,Embase,Cochrane,VIP,CBM,WanFang,and CNKI databases to search relevant articles published from database inception to May 2024. Inclusion and exclusion criteria were developed,and 97 relevant articles were selected by reading titles,abstracts,and full texts. RESULTS AND CONCLUSION:Natural products such as polyphenols can directly scavenge oxidative free radicals by the phenol hydroxyl group in their structure,and alleviate oxidative stress after spinal cord injury. Some natural products can enhance the activity of relevant antioxidant enzymes in vivo and reduce oxidative stress by regulating certain signal transduction pathways. Some natural products can reduce oxidative stress after spinal cord injury by enhancing autophagy. The use of natural products to regulate oxidative stress may become an effective tool for the future clinical treatment of spinal cord injury.

Objective:To explore the spatial correlation between gray matter volume (GMV) changes and neurotransmitter receptors/transporters in patients with first-episode schizophrenia (FES) .Methods:Fifty-four FES patients(FES group) and fifty-nine healthy controls (HC group) were selected from June 2014 to May 2020 in the Psychiatry Department of Renmin Hospital of Wuhan University. Structural magnetic resonance imaging (sMRI) was conducted on all subjects. Differences of GMV were compared across 400 cortical regions and 32 subcortical regions. Based on the positron emission tomography(PET) data from Neuromaps, which provides the density of 19 different neurotransmitter receptors and transporters, Spearman correlation analysis was performed to evaluate the spatial correlation between GMV changes and neurotransmitter systems.Results:Compared to the HC group, FES group exhibited significant GMV reductions in widespread cortical (90/400) and subcortical (6/32) regions (all FDR-corrected P<0.05). The effect size of GMV reduction (Cohen’s d) showed significant positive correlations with the density of 5-hydroxytryptamine 1a(5HT1a) ( r=0.400, Pspin=0.002), γ-aminobutyric acid type A receptor(GABA A)( r=0.307, Pspin=0.002), and metabotropic glutamate receptor 5(mGluR5) ( r=0.275, Pspin=0.020) receptors (all FDR-corrected P<0.05). Conclusion:GMV reductions in a wide range of brain regions existed in patients with FES. There are significant correlations between 5HT1a, GABA A and mGluR5 receptors and gray matter reduction in patients with FES. The disorder of these neurotransmitter receptors may be the potential neurobiological mechanism of gray matter structural abnormalities in the early stage of schizophrenia.