BACKGROUND:Autophagy,as a key regulatory mechanism of cell development,plays an important role in different stages of embryonic development.The mechanism of how autophagy regulates embryonic development through histone modifications is currently unclear.OBJECTIVE:To investigate the effect of autophagy on trimethylation of lysine 4 on histone H3(H3K4me3)modification in embryos and its effect on embryonic development.METHODS:Mouse fertilized eggs were divided into control and autophagy inhibitor-treated groups(chloroquine phosphate-treated group and 3-methyladenine-treated group),and cultured in vitro to different periods of time,and were then classified as early 2-cell embryos,middle 2-cell embryos,late 2-cell embryos,4-cell embryos,8-cell embryos,morula stage,and blastocyst stage.Levels of reactive oxygen species,autophagy marker proteins LC3B and P62,DNA loss marker γH2AX,and H3K4me3 were analyzed by immunofluorescence assay in late 2-cell embryos of each group.Changes in H3K4me3 modification in late 2-cell embryos of each group were detected by CUT&Tag.RESULTS AND CONCLUSION:(1)Autophagy inhibition caused embryo development arrest.(2)There was no significant difference in reactive oxygen species and γH2AX between the autophagy inhibitor-treated groups and control group.(3)H3K4me3 levels were significantly elevated in the autophagy inhibitor-treated group compared with the control group.(4)CUT&Tag results showed a significantly increased H3K4me3 peaks on the proximal promoter region of the genes after autophagy inhibition and an increase of H3K4me3-specific modification genes.These findings suggest that autophagy may affect embryonic development by regulating the level of H3K4me3 modification.

[摘 要] 树突状细胞（DC）作为抗肿瘤免疫应答的核心启动者与调控者，已成为肿瘤免疫治疗的重要靶点。DC疫苗等靶向DC的免疫治疗策略在抗肿瘤治疗中展现出独特优势，但仍存在抗原提呈效率不足、免疫抑制微环境抵抗、功能特异性调控困难等瓶颈问题。肿瘤微环境（TME）中的DC存在高度异质性，不同DC亚群在分化发育、免疫调控及效应转归方面呈现复杂的多样性。解析DC亚群的精确表型与功能机制对于开发新型DC靶向性免疫治疗策略具有重要意义。TME中募集的经典DC、浆细胞样DC、单核细胞来源DC能与肿瘤浸润免疫细胞和微环境中的非免疫细胞（包括肿瘤细胞、成纤维细胞、内皮细胞等）相互作用激活抗肿瘤免疫应答，而TME还会通过转录调控、表观遗传调控、代谢重塑等多种方式抑制DC招募和抗原提呈能力，甚至诱导其向耐受性DC转化。值得注意的是，新近发现的富含免疫调节分子的成熟DC具有双向免疫调控作用，其起源路径和免疫调控网络仍有待深入研究。随着单细胞分析与空间组学分析等技术的发展，有望在单细胞分辨率下系统解析DC亚群的功能多样性及其与TME的时空相互作用网络，为开发下一代精准免疫治疗策略提供新靶点和新思路。

Neurocritical care involves complex pathophysiological mechanisms, and its incidence is higher, injuries are more severe, and treatment is more challenging in high-altitude environments. This consensus, based on the latest domestic and international evidence-based medical data, establishes a standardized, goal-oriented framework for neurocritical care management applicable in high-altitude regions and nationwide. The consensus was developed following international standards for evidence quality assessment and underwent two rounds of Delphi expert consultation, resulting in 32 recommendation statements covering three parts: management systems, monitoring and assessment, and core strategies. Key updates include: advocating for the establishment of independent neurocritical care units and implementing precise tiered diagnosis and treatment based on the "Five Differences in Critical Care" concept; constructing a "trinity" multimodal brain monitoring system centered on cerebral blood flow, cerebral oxygenation, and brain function, emphasizing routine bedside transcranial Doppler ultrasound, cerebral oximetry, and continuous electroencephalography monitoring; shifting management strategies from mild hypothermia therapy to targeted temperature management, and defining the "446" target management pathway for the supercritical stage; emphasizing the assessment of static and dynamic cerebrovascular autoregulation functions through multimodal methods to achieve individualized optimal mean arterial pressure management; elevating cerebrospinal fluid management goals to the level of "glymphatic system" function maintenance; implementing a multidisciplinary collaborative, whole-process management model focusing on patients' long-term neurological functional outcomes; de-escalation criteria include multidimensional indicators such as recovery of brain structure, restoration of cerebrovascular autoregulation, improvement in cerebrospinal fluid dynamics, and reduction in biomarker levels; and integrating cutting-edge technologies like artificial intelligence into post-critical care management and rehabilitation planning. This consensus systematically integrates the entire process of neurocritical care management, reflecting the modern connotation of goal-oriented, dynamic, and multimodal integration in neurocritical care medicine. It aims to adapt to new trends such as deepening understanding of pathophysiological mechanisms, the integration of medicine and engineering, and the empowerment of artificial intelligence, thereby further advancing the discipline of critical care medicine.

BACKGROUND:Autophagy,as a key regulatory mechanism of cell development,plays an important role in different stages of embryonic development.The mechanism of how autophagy regulates embryonic development through histone modifications is currently unclear.OBJECTIVE:To investigate the effect of autophagy on trimethylation of lysine 4 on histone H3(H3K4me3)modification in embryos and its effect on embryonic development.METHODS:Mouse fertilized eggs were divided into control and autophagy inhibitor-treated groups(chloroquine phosphate-treated group and 3-methyladenine-treated group),and cultured in vitro to different periods of time,and were then classified as early 2-cell embryos,middle 2-cell embryos,late 2-cell embryos,4-cell embryos,8-cell embryos,morula stage,and blastocyst stage.Levels of reactive oxygen species,autophagy marker proteins LC3B and P62,DNA loss marker γH2AX,and H3K4me3 were analyzed by immunofluorescence assay in late 2-cell embryos of each group.Changes in H3K4me3 modification in late 2-cell embryos of each group were detected by CUT&Tag.RESULTS AND CONCLUSION:(1)Autophagy inhibition caused embryo development arrest.(2)There was no significant difference in reactive oxygen species and γH2AX between the autophagy inhibitor-treated groups and control group.(3)H3K4me3 levels were significantly elevated in the autophagy inhibitor-treated group compared with the control group.(4)CUT&Tag results showed a significantly increased H3K4me3 peaks on the proximal promoter region of the genes after autophagy inhibition and an increase of H3K4me3-specific modification genes.These findings suggest that autophagy may affect embryonic development by regulating the level of H3K4me3 modification.

ObjectiveTo investigate the therapeutic effect and mechanism of modified Chunzetang on bladder fibrosis and detrusor function in rats with neurogenic bladder urinary retention induced by spinal cord injury. MethodsIn this study， an improved Hassan Shaker spinal cord transection method was used to establish a model of neurogenic bladder urinary retention induced by spinal cord injury， and rats with a spinal cord injury behavior score of 0 were selected for follow-up experiments. The selected rats were randomly divided into a model group （normal saline gavage）， low-dose traditional Chinese medicine （TCM） group （gavage of 14.4 g·kg^-1 modified Chunzetang）， high-dose TCM group （gavage of 28.8 g·kg^-1 modified Chunzetang）， positive drug group ［intraperitoneal injection of 0.05 g·kg^-1 nuclear transcription factor-κB （NF-κB） inhibitor pyrrolidine dithiocarbamate （PDTC）］， and combination group （intraperitoneal injection of 0.05 g·kg^-1 PDTC + gavage of 28.8 g·kg^-1 modified Chunzetang）. The rats in these groups were administrated with corresponding drugs once a day for four weeks. The BL-420s biofunction acquisition system was used in the experiment to calculate the urodynamic indexes， and the isolated bladder was quickly weighed. The detrusor traction experiment was used to record the minimum bladder contraction tension and frequency in each group. The pathological morphology and tissue fibrosis of detrusor in each group observed by Hematoxycin-eosin （HE） staining and Masson staining were compared. The expression level of α-smooth muscle actin （α-SMA） was detected by immunohistochemistry. Western blot was used to detect the protein expression of NF-κB p65， nuclear transcription factor-κB suppressor protein α （IκBα）， transforming growth factor-β₁ （TGF-β₁）， type Ⅰ collagen （ColⅠ）， and type Ⅲ collagen （ColⅢ） in bladder tissue of rats in each group. Enzyme-linked immunosorbent assay （ELISA） was used to detect the changes in serum levels of IL-6， IL-1β， and TNF-α. ResultsCompared with that in the sham operation group， the pressure at the urinary leakage point in the model group decreased （P<0.01）， and the bladder mass， bladder contractile tension， maximum bladder capacity， and bladder compliance increased （P<0.05，P<0.01）. HE staining showed that the arrangement of bladder epithelial cells was disordered， and the pathological manifestations such as mucosa and myometria neutrophil infiltration were obvious. The lamina propria structure was destroyed， and the muscle fiber arrangement was disordered. The interstitial widening and tissue edema were obvious. Masson staining showed that the bladder wall of the model group had more collagen fiber deposition， and the degree of detrusor fibrosis was more severe. The content of detrusor in the visual field was reduced. At the same time， the protein expressions of NF-κB p65， TGF-β₁， IκBα， ColⅠ， and ColⅢ in bladder tissue of rats in the model group were significantly increased （P<0.01）， and the serum levels of IL-6， IL-1β， and TNF-α were significantly increased （P<0.05）. Compared with that in the model group， the pressure at the urinary leakage point in the modified Chunzetang and positive drug groups was increased （P<0.05）， and the wet bladder weight， minimum bladder contractile tension， maximum bladder capacity， and bladder compliance were restored （P<0.05， P<0.01）. HE and Masson showed that the bladder epithelial cells were relatively neatly arranged， and the structure of the bladder lamina propria was relatively stable. The detrusor bundles were arranged in an orderly manner， and the interstitium was narrow. The degree of tissue edema was relatively low， and the degree of bladder detrusor fibrosis in the modified Chunzetang and positive drug groups was reduced， while the degree of bladder detrusor fibrosis in the positive drug group and combination groups was not obvious. The results of Western blot showed that the expression of NF-κB p65， IκBα， TGF-β₁， ColⅠ， and ColⅢ in bladder tissue， as well as the serum levels of IL-6， IL-1β， and TNF-α in modified Chunzetang and positive drug groups were significantly lower， and the expression of bladder tissue-related proteins and the serum levels of IL-6， IL-1β， and TNF-α in the TCM groups decreased significantly with the increase in dose （P<0.05）. The results of immunohistochemistry suggested that modified Chunzetang could fully affect the expression of α-SMA in bladder tissue. ConclusionModified Chunzetang can inhibit collagen deposition in bladder tissue of rats with urinary retention induced by spinal cord injury， delay the occurrence and development of bladder fibrosis， and protect the normal contractile function of bladder detrusor， and its mechanism may be related to inhibiting the NF-κB/TGF-β₁ signaling pathway， reducing the production of NF-κB p65， IκBα， TGF-β₁， ColⅠ， ColⅢ， and other related proteins， and protecting the muscle strength of detrusor.

Non-alcoholic fatty liver disease （NAFLD） is a chronic liver disease closely related to metabolism， which is mainly characterized by abnormal lipid deposition in hepatocytes. In recent years， with the increasing prevalence of obesity and metabolic syndrome， NAFLD has become one of the most common chronic diseases in the world. The pathogenesis of NAFLD is complex and varied， involving the cross-regulation of multiple signaling pathways such as glucose-lipid metabolism， oxidative stress， and inflammation. The TLR4 signaling pathway plays a key role in the development and progression of NAFLD， and abnormal activation of this pathway accelerates the deterioration of NAFLD by promoting the release of pro-inflammatory cytokines， inducing oxidative stress， and exacerbating insulin resistance. Studies have shown that traditional Chinese medicine （TCM） can regulate the TLR4 signaling pathway to alleviate the symptoms and pathological features of NAFLD. The present review summarizes the experimental research progress in the TCM regulation of the TLR4 signaling pathway in treating NAFLD in the past 5 years， covering a wide range of TCM active ingredients （such as polysaccharides， terpenoids， alkaloids， flavonoids） and compound prescriptions. The active ingredients and compound prescriptions of TCM can effectively ameliorate lipid metabolism disorders， reduce insulin resistance， regulate intestinal flora， and inhibit inflammation and oxidative stress by regulating the TLR4 signaling pathway via multiple targets and pathways， thus slowing down the progression of NAFLD. Through in-depth analysis of the pathological mechanisms of NAFLD and exploration of the potential of TLR4 signaling pathway as a therapeutic target， we can provide theoretical support for the application of TCM in the treatment of NAFLD， as well as new perspectives and directions for future clinical research and new drug development， thereby promoting the innovation and development of therapeutic strategies for NAFLD.

Objective:To investigate the effects of lezumab adjuvant therapy on the degree of retinal vasculopathy and immune response in the treatment of macular central diabetic retinopathy (DR).Methods:From July 2022 to December 2023, 120 patients with DR in macular central who received treatment in the Xi′an People′s Hospital (Xi′an Fourth Hospital) were retrospectively selected and divided into two groups according to the treatment methods: the observation group (60 cases, lezumab combined with sitagliptin), and the control group (60 cases, sitagliptin). Patients′glucose metabolism, visual acuity, degree of retinopathy, inflammation [tumor necrosis factor-alpha (TNF-α), interleukin-2 (IL-2), interleukin-10 (IL-10)] and angiogenesis related factors [serum vascular endothelial growth factor (VEGF), midkine (MK), 5′-nucleotidase (CD73)] were evaluated, and the curative efficacy of the two groups was compared. The security of the two schemes was compared.Results:Compared with the control group, the serum concentrations of MK, CD73, TNF-α and VEGF were lower in the observation group after treatment: (1.44 ± 0.06) ng/L vs. (1.67 ± 0.11) ng/L, (1.10 ± 0.27) ng/L vs. (1.31 ± 0.26) ng/L, (11.62 ± 0.89) ng/L vs. (15.96 ± 4.42) ng/L, (84.07 ± 27.07) ng/L vs. (100.72 ± 16.05) ng/L, while the concentration of IL-10 was higher: (65.65 ± 8.68) ng/L vs. (60.02 ± 5.07) ng/L, with statistically significant differences ( P<0.05). There were no statistically significant differences in fasting blood glucose (FBG) and 2 h postprandial blood glucose between two groups before and after treatment ( P>0.05). After treatment, the macular thickness and visual field gray value in the observation group were lower than those in the control group: (302.81 ± 77.08) μm vs. (336.44 ± 10.35) μm, (1.55 ± 0.43)% vs. (2.09 ± 0.51)% ( P<0.05). After 3 months of treatment, the visual acuity in the observation group was higher than that in the control group: 0.493 ± 0.103 vs. 0.439 ± 0.084 ( P<0.05). No serious adverse reactions occurred in both groups. Conclusions:Lezumab assisted sitagliptin has a significant effect in the treatment of DR, which can reduce the degree of lesions, improve vision, and reduce the levels of inflammation and angiogenesis related factors.

Objective:To explore the application effect of 3D technology-assisted teaching based on "four-in-one" flipped classroom in clinical teaching of neurosurgery, and provide a basis for optimizing the medical education mode.Methods:A total of 50 students from the Second Clinical College were selected between June 2020 and June 2024, including eight-year program medical students and postgraduate neurosurgery students. The control group ( n=25) received the conventional teaching mode. The experimental group ( n=25) was taught using the "four-in-one" flipped classroom combined with 3D technology, including characteristic textbooks with 3D model drawings, Internet platforms (video libraries and virtual simulation modules), mobile interactive terminals (real-time question and answer), and virtual simulation technology. Effectiveness was evaluated through theoretical assessment (e.g., neuroanatomy and clinical application ability, with a total score of 100 points), operational assessment (e.g., surgical design and aseptic concept, with a total score of 100 points), and a teaching quality questionnaire. Independent samples t-test was conducted using SPSS 22.0. Results:The total score of theoretical assessment was higher in the experimental group than that in the control group [(86.52±5.21) vs. (73.56±6.32), P<0.001], with the largest difference observed in case analysis questions [(26.03±3.65) vs. (22.22±3.50), P=0.001]. In the operational assessment, the experimental group performed better in "surgical process design" [(26.30±4.14) vs. (21.44±3.45), P<0.001] and "aseptic concept" [(8.18±0.98) vs. (6.64±0.79), P<0.001]. The teaching quality questionnaire showed that the experimental group scored higher in "clinical practice skill enhancement" [(23.13±1.39) vs. (21.45±1.86), P=0.001] and "self-directed learning motivation" [(21.84±1.60) vs. (19.75±1.45), P<0.001]. Conclusions:The combination of "four-in-one" flipped classroom and 3D technology can significantly improve the teaching effectiveness of neurosurgery, especially in the cultivation of clinical thinking and practical abilities. This approach is worth promoting.

Ischemic stroke,a major public health challenge,is marked by high incidence,disability,and mortality rates.The pathophysiology of stroke is complex,and inflammation reaction plays a crucial role throughout the process.In recent years,anti-inflammatory treatment has become a major focus in researches of ischemic stroke.Current preclinical and clinical investigations have revealed the potential of various anti-inflammatory strategies for ischemic stroke.However,which patient populations benefit from existing drugs are not yet elucidated.Additionally,drugs targeting innate immune cascades have become a new research hot spot.This article reviewed the latest clinical advances in anti-inflammatory therapy for ischemic stroke and discusses its promising prospects,aiming to provide novel clinical application strategies and future research directions.

Objective To explore the diagnostic value of the radiomic model based on panoramic radiographs in differentiating single or multiple root canals in mandibular first premolars.Methods A retrospective analysis was conducted on 68 patients with CBCT and panoramic radiographs from the affiliated Stomatological Hospital of Nanjing Medical University between July 2022 and November 2023.One hundred and thirty one mandibular first premolars(105 single-root and 26 multiple-root canals)were included in this study.Using stratified random sampling,the mandibular first premolars were divided into a training cohort(104 teeth)and a test cohort(27 teeth)at a ratio of 8∶2.CBCT was used to confirm the root canal configuration,and radiomic features were extracted from panoramic radio-graphs.After feature screening,radiomic models were constructed using extratrees,LightGBM,logistic regression(LR),multi-layer perceptron(MLP),random forest,support vector machine(SVM),and XGBoost.The optimal radiomic model was selected based on receiver operating characteristic(ROC)curve,accuracy,sensitivity,specificity,positive predictive value(PPV),negative predictive value(NPV),and decision curve analysis(DCA).Results A total of 107 radiomicfeatures were extracted from panoramic radiographs.After feature filtering,4 radiomic features were ultimately selected using the Lasso model.The XGBoost model showed higher predictive efficiency and stability,with an AUC of 0.962 in the training cohort and 0.770 in the test cohort.DCA curves also in-dicated that the XGBoost model was optimal.Conclusion The radiomic model based on panoramic radiographs demonstrates good per-formance in differentiating single or multiple root canals in mandibular first premolars.