[摘 要] 目的：探讨胃肝样腺癌（HAS）的临床病理特征与免疫微环境异质性，筛选预后标志物，阐释其高侵袭性与疗效差的机制，为精准诊疗策略提供理论依据。方法:​​ 回顾性收集2013年1月至2025年5月期间海军军医大学第一附属医院与第二附属医院收治的65例HAS患者的临床信息及病理资料。采用免疫组织化学技术检测HAS​​肝样分化标志物、神经内分泌标志物​​等分子表达情况，通过Kaplan-Meier生存分析法明确与预后相关的靶点。利用多色免疫荧光技术鉴定肿瘤区域内免疫细胞亚群分布情况，以阐明其免疫微环境特征。结果：65例患者中，男性54例（83.1%），女性11例（16.9%），中位年龄68岁。肿瘤好发于贲门（40%），其次为胃窦（32.3%）和胃体（27.7%）。中位随访时间23.18个月，15例患者死亡，46例生存，4例失访。HAS的胃镜及手术标本大体观呈灰白色实性质硬肿物，镜下可见中低分化胃腺癌与肝细胞癌（HCC）样分化区交错分布。表达神经内分泌相关分子的HAS患者呈现出更多的淋巴结转移数量及更短的总生存期。免疫微环境解析显示，HAS总体缺乏免疫细胞浸润，呈现“冷肿瘤”特征；免疫细胞主要聚集于胃癌腺体周围区域，而在HCC样分化区域罕见淋巴细胞浸润。结论：HAS侵袭性强，根治性手术是主要治疗手段；神经内分泌转化提示不良预后，是个体化治疗的关键标志；免疫细胞浸润缺乏，可能是其免疫治疗响应不佳的原因。

OBJECTIVE To explore the mechanism of action of Qingre huatan huoxue decoction against atherosclerosis （AS）based on macrophage polarization. METHODS Using atorvastatin served as the positive control， the drug-containing serum of the Qingre huatan huoxue decoction was prepared to treat RAW264.7 macrophages. Macrophage viability， apoptosis rate， and the fluorescence intensities of CD86 and CD206 were measured， along with the levels of tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β）. Apolipoprotei n E-deficient （ApoE －/－ ） mice （AS model mice） fed with a high-fat diet were randomly assigned to model group， atorvastatin group （2.6 mg/kg）， and low-， medium- and high-dose groups of Qingre huatan huoxue decoction （90， 180， 360 mg/kg）， respectively. C57BL/6J mice fed with a standard diet served as the normal control group， with 10 mice per group. The treatment group mice were administered the corresponding drugs intragastrically， once daily， for 8 consecutive weeks. Serum levels of TNF-α and IL-1β were measured in all groups. Lipid deposition in the aorta （assessed by the percentage of plaque in the entire aorta and aortic root） and morphological changes in the aortic root were observed. Expression levels of CD86 and CD206 in aortic tissue， as well as the protein expression levels of inducible nitric oxide synthase （iNOS）， arginase-1 （Arg-1）， AMP-activated protein kinase （AMPK）， phosphorylated AMPK （p-AMPK）， and peroxisome proliferator-activated receptor γ （PPAR-γ） in aortic tissues were all detected. RESULTS Cell experiment results showed that， at concentrations of 5-100 μg/mL， the drug-containing serum of the Qingre huatan huoxue decoction significantly increased RAW264.7 cell viability （ P ＜0.05）. The drug-containing serum of the Qingre huatan huoxue decoction at concentrations of 10， 50， and 100 μg/mL， along with atorvastatin， significantly reduced apoptosis rates， CD86 fluorescence intensity， and TNF-α and IL-1β levels in RAW264.7 cells， while markedly enhancing CD206 fluorescence intensity （ P ＜0.05）. Animal experiment results showed that， compared with the model group， all dosage groups of Qingre huatan huoxue decoction and the atorvastatin group showed significantly reduced/down-regulated levels of TNF-α and IL-1β in serum， along with decreased aortic total and root plaque percentages， CD86 expression， and iNOS protein expression. CD206 expression and Arg-1， p-AMPK/AMPK， PPAR-γ protein expression were significantly up-regulated （ P ＜0.05）. Pathological morphology of the aorta showed varying degrees of improvement. CONCLUSIONS The formula of Qingre huatan huoxue decoction exerts its anti-AS effects by regulating macrophage polarization， increasing the proportion of M2 macrophages， thereby effectively inhibiting AS plaque formation and reducing inflammatory responses.

Objective: To investigate whether membrane vesicles (MVs) of Streptococcus mutans （S.mutans） contain autoinducer-2 (AI-2) and to preliminarily explore the effects of these MVs on the growth and biofilm formation of S. mutans. Methods: MVs were isolated from the S. mutans UA159 strain using differential centrifugation. The isolated MVs were characterized by nanoparticle tracking analysis for particle size and concentration and observed by transmission electron microscopy. The presence of AI-2 was identified using the Vibrio harveyi BB170 bioluminescence assay: the BB170 diluent was supplemented with AB medium (control group), MV extract (MVs group), pre-ultrafiltration supernatant (Sup group), or post-ultrafiltration supernatant (Sup-af group). The effects of MVs on growth and biofilm formation were assessed using the S.mutans UA159 strain or a luxS deletion mutant as the control group, compared with experimental groups stimulated with gradient concentrations of MVs (MVs-2.0E+7, MVs-2.0E+8, and MVs-2.0E+9 groups). Growth curves, MTT assay, and colony-forming unit (CFU) counts were used to determine changes in growth capacity. Biofilm formation was evaluated using crystal violet staining, confocal laser scanning microscopy, and the anthrone method for polysaccharide quantification. Results: Enriched S. mutans MVs were successfully obtained, with an average particle size of approximately 94.19 nm and a concentration of 1.87E+11 particles/mL. The bioluminescence assay showed that the luminescence intensity of the Sup group was higher than that of the Sup-af group, and the MVs group exhibited higher intensity than the control group. Assessments via growth curves, MTT assay, and CFU counts indicated no significant differences in the growth capacity of the various S. mutans strains after treatment with different concentrations of MVs. Crystal violet staining quantification and confocal laser scanning microscopy observations revealed that high-concentration MV treatment (2.0E+9 particles/mL group) resulted in lower biofilm mass compared to the control. The anthrone method showed that the production of both water-soluble and water-insoluble polysaccharides was significantly lower in the high-concentration MV group than in the control. Conclusion S. mutans MVs contain the quorum sensing signal molecule AI-2. These MVs do not significantly affect the growth of S. mutans, but they can regulate biofilm formation and exhibit an inhibitory effect at high concentrations.

Radiotherapy is a crucial treatment modality for head and neck tumors. However, while effectively killing tumor cells, it significantly disrupts the homeostasis of the oral microecology, which is closely associated with various complications such as radiation-induced oral mucositis. Literature review indicates that as radiotherapy doses accumulate and treatment durations extend, the richness and diversity of the oral microbiota show a declining trend, with the genus Streptococcus decreasing most markedly. In contrast, radiotherapy selectively promotes the proliferation of bacterial phyla such as Proteobacteria and Bacteroidetes, which are rich in opportunistic pathogens. Mechanistically, radiotherapy activates the nuclear factor-kappa B pathway, triggering chronic inflammation and oxidative stress, damaging the epithelial barrier, suppressing local immunity, and causing damage to organs such as the salivary glands. It can also induce systemic diseases via the oral-gut axis, forming a multi-level, interconnected pathogenic network. In terms of interventions, treatment strategies including probiotics and prebiotics have shown promising efficacy against side effects such as radiation-induced oral mucositis. Saliva-based oral microbiota transplantation is an emerging strategy that is expected to become widely utilized for restoring oral microecological balance. Existing interventions provide preliminary pathways for clinical practice, but this field still faces several key scientific questions. The association between oral microecology and systemic diseases remains largely correlative, lacking causal evidence. Furthermore, critical parameters for oral microbiota transplantation, such as donor screening criteria, transplantation protocols, and long-term safety, are not yet well-defined. Therefore, future research should focus on conducting large-scale clinical trials to establish standardized protocols and safety evaluation systems for oral microecological interventions, and explore combined treatment therapies such as probiotics, prebiotics, and microbiota transplantation to advance the development of personalized precision modulation. These will enable more effective management of radiotherapy-induced oral microecological dysbiosis and improve treatment outcomes and quality of life for patients with head and neck tumors.

The choroidal vascularity index(CVI)is a quantitative structural parameter derived from optical coherence tomography(OCT)images. It is defined as the ratio of the luminal area to the total choroidal area, which characterizes alterations in the structural proportion between choroidal vascular lumens and stroma, and indirectly indicates changes in choroidal status to a certain extent. With the development of enhanced depth imaging optical coherence tomography(EDI-OCT), CVI has been increasingly applied in ophthalmic research and clinical practice.Measurements of CVI exhibit excellent repeatability and reliability. Combined with the noninvasive nature of EDI-OCT, CVI is an extremely valuable assessment tool for clinical ophthalmic research and routine clinical practice. CVI can quantify proportional changes between the choroidal luminal area and total choroidal area, and better reflects relative variations in choroidal vascular and stromal components compared with choroidal thickness alone. In recent years, CVI has been widely applied to multiple ocular diseases including age-related macular degeneration(ARMD), central serous chorioretinopathy(CSC), diabetic retinopathy(DR), and high myopia. In addition to the diseases themselves, various ocular surgeries and procedures can also affect CVI. Current evidence suggests that changes in CVI may be associated with factors such as postoperative inflammatory response, choroidal hemodynamic alterations, vascular permeability changes, and intraocular pressure fluctuations. However, discrepancies remain among studies regarding the direction and duration of these changes and their relationship with visual outcomes. This review summarizes the effects of ocular surgeries and procedures on CVI and further discusses its clinical interpretation, existing controversies, and future research directions, aiming to provide a reference for perioperative assessment of choroidal structural changes and prognostic evaluation.

Photoacoustic microscopy(PAM)is an emerging, non-invasive, in vivo imaging modality that merges optical and acoustic principles. It offers high-resolution and high-contrast visualization of various ocular tissue structures and functional information, making it suitable for studying a wide range of ophthalmic diseases such as corneal neovascularization, macular degeneration, and diabetic retinopathy. The multi-wavelength illumination capability of PAM makes it particularly valuable for early disease screening and dynamic physiological monitoring. In stem cell tracking, PAM enables the dynamic monitoring of transplanted cells through contrast agent labeling. Moreover, when combined with multimodal imaging techniques like optical coherence tomography(OCT), PAM can enhance the detection accuracy and diagnostic capacity for ocular diseases. However, PAM still requires optimization in terms of imaging speed and contrast agent safety. This review summarizes the fundamental principles and development of PAM, explores its applications in specific ophthalmic diseases, and analyzes the challenges and optimization directions from animal experiments to clinical applications. PAM holds great promise for playing a more significant role in ophthalmic diagnosis and treatment.

BACKGROUND:Platelet-rich fibrin(PRF)has many advantages,such as simple preparation,low production cost,and high safety,and has been widely used in the study of bone defect repair in oral and maxillofacial surgery,but there are problems such as too fast degradation rate and short release time of growth factors. OBJECTIVE:PRF was loaded into gelatin methacryloyl(GelMA)hydrogel and its osteogenic properties were analyzed by in vivo and in vitro experiments. METHODS:(1)New Zealand white rabbit venous blood was extracted to prepare PRF.GelMA hydrogels containing 0,0.05,0.075,and 0.1 g PRF were prepared,respectively,and were recorded as GelMA,GelMA/PRF-0.05,GelMA/PRF-0.075,and GelMA/PRF-0.1,respectively,to characterize the micromorphology and in vitro slow-release properties of the hydrogels.(2)Four kinds of hydrogels were co-cultured with MC3T3-E1 cells,respectively,and the cell proliferation activity was detected with the single cultured cells as the control.After osteogenic induction,alkaline phosphatase activity,mineralization ability,mRNA and protein expression levels of osteogenic genes(osteocalcin,osteopontin,RUNX2),ERK1/2-p38 MAPK pathway protein mRNA and protein expression levels were detected.(3)Fifteen New Zealand white rabbits were taken.Four full-layer bone defects of 8 mm diameter were prepared in the skull of each rabbit,one of which was implanted without any material(blank control group),and the other three were implanted with GelMA hydrogel,PRF,and GelMA/PRF-0.1 hydrogel,respectively.The bone defect was detected by Micro-CT and bone morphology was observed at 4,8,and 12 weeks after operation. RESULTS AND CONCLUSION:(1)Scanning electron microscopy observed that all the hydrogels of the four groups had honeycomb pore structure,and the pore size of the hydrogels decreased slightly with the increase of PRF content,but there was no significant difference between the groups.The three groups of GelMA/PRF hydrogel could release transforming growth factor β1 and insulin-like growth factor 1 at a certain rate,and the cumulative release of transforming growth factor β1 and insulin-like growth factor 1 increased significantly with the extension of time.(2)CCK-8 assay and live/dead staining showed that GelMA/PRF hydrogel could promote the proliferation of MC3T3-E1 cells.The results of alkaline phosphatase staining,alizarin red staining,and osteogenic gene detection showed that GelMA/PRF hydrogel could promote the osteogenic differentiation of MC3T3-E1 cells,and inhibit the expression of ERK1/2-p38 MAPK pathway protein,and showed a PRF content dependence.(3)Micro-CT scan showed that the bone mineral density and bone volume fraction in the bone defect of GelMA/PRF-0.1 hydrogel group were higher than those in the other three groups(P<0.05).Hematoxylin-eosin staining showed that compared with the other three groups,GelMA/PRF-0.1 hydrogel group had faster and more mature new bone formation at the bone defect.(4)These findings indicate that GelMA/PRF hydrogel has good osteogenic activity both in vivo and in vitro,which may be related to inhibiting the expression of ERK1/2-p38 MAPK pathway protein.

Objective To investigate the influencing factors for stroke-associated pneumonia(SAP)in acute ischemic stroke(AIS)patients after endovascular treatment(EVT).Methods A retrospective case-control trial was conducted on 426 AIS patients with large vessel occlusion(LVO)in anterior circulation admitted in the neurological departments from First Affiliated Hospital of Army Medical University and Zigong Third People's Hospital during January 2017 and April 2021.Based on SAP occurrence or not,they were divided into an SAP group and a non-SAP group.Demographic information(gender and age),TOAST stroke subtypes(large artery atherosclerosis type,cardiac embolism type,others),vascular risk factors(hypertension,hyperlipidemia,diabetes,atrial fibrillation,smoking,prior stroke history,smoking),and post-onset clinical data[dysphagia,LDL cholesterol,white blood cells,neutrophils,baseline and postoperative NIHSS scores,endovascular outcomes(mTICI grade 2b or 3),90-day good prognosis(mRS 0-1)]were collected and compared between the 2 groups.Multivariate logistic regression analysis was performed using the parameters with P<0.1 in univariate analysis as independent variables to investigate factors influencing SAP occurrence after EVT in AIS patients.Results Among the 426 participants,SAP occurred in 194 cases(45.5%).Multivariate logistic regression analysis revealed that admission white blood cell count(OR=1.125,95%CI:1.043～1.213,P=0.000 2),postoperative NIHSS score(OR=1.019,95%CI:1.001～1.037,P=0.041),and male(OR=1.687,95%CI:1.078～2.638,P=0.022)were associated with SAP occurrence after EVT in AIS patients.Conclusion Higher admission white blood cell count,elevated postoperative NIHSS score,and male gender are risk factors for SAP in AIS patients after EVT.These risk factors should be focused on clinical practice to control SAP incidence.

Objective To investigate the role and mechanism of ferroptosis in cochlear hair cell injury induced by exposure to ammonium 2,3,3,3-tetrafluoro-2-(heptafluoropropoxy)propanoate(GenX).Methods Mouse cochlear hair cell line HEI-OC1 was assigned to control,GenX,ferrostatin-1(Fer-1),and GenX+Fer-1 groups.CCK-8 assay was used to assess the cytotoxicity of GenX and the rescue effect of Fer-1 co-treatment.Western blotting and qRT-PCR were employed to measure the protein and transcriptional expression of ferroptosis markers,cochlear function indicators,blood labyrinth barrier markers,and ferroptosis-related pathway.FerroOrange,Bodipy(C11),mitochondrial membrane potential assay kit(JC-1),and adenosine triphosphate(ATP)assay were applied to detect Fe2? accumulation,lipid peroxidation,mitochondrial membrane dysfunction,and cellular ATP levels,respectively.Results Exposure to 200 μmol/L GenX for 12 h significantly reduced the viability of HEI-OC1 cells(P<0.01),down-regulated the protein levels of glutathione peroxidase 4(Gpx4),solute carrier family 7 member 11(Slc7a11),and ferroptosis suppressor protein 1(Fsp1)(P<0.05),whereas up-regulated acyl-coa synthetase long-chain family member 4(Acsl4)(P<0.01),and decreased the expression of cochlear hair-cell function genes and blood labyrinth barrier genes(P<0.05).These changes were accompanied with Fe2+accumulation,elevated lipid peroxidation,mitochondrial membrane damage,and reduced ATP production(P<0.001).Addition of Fer-1 restored cell viability(P<0.05),restored the expression of ferroptosis related proteins(P<0.05),and improved the expression of several hair-cell function and blood labyrinth barrier genes(P<0.05).In parallel,the GenX+Fer-1 group exhibited reduced Fe2? accumulation,lower lipid peroxidation,attenuated mitochondrial membrane damage,and increased ATP level(P<0.001).Conclusion GenX induces iron-metabolism dysregulation and lipid peroxidation,and then leads to differentiation impairment of cochlear hair cells and barrier functions via the ferroptotic System Xc?-Gpx4-Fsp1 axis.

Objective To explore the pharmacokinetic characteristics of sufentanil in individuals with normal body mass index(BMI),overweight BMI,and different CYP3A4/5 enzyme genotypes.Methods The patients receiving laparoscopic surgery under general anesthesia in the First Affiliated Hospital of Army Medical University from November 2020 to September 2021 were prospectively recruited in this study.Before the operation,the oral swabs were collected from all the patients for genotyping using the human CYP3A4/5 gene kit.Based on the potential impact of combination of their polymorphisms on sufentanil metabolism and the proportion of different genotype combinations of CYP3A4/5 enzymes,the patients were divided into groups I(3A4 homozygous mutation or 3A4 heterozygous mutation+3A5 homozygous mutation),II(3A4 heterozygous mutation+3A5 heterozygous mutation),and III(3A4 wild type or 3A4 heterozygous mutation+3A5 wild type).According to their BMI,they were also assigned into a normal body weight group(18.5～24.0 kg/m2)and an overweight group(24～<28 kg/m2),and the differences in drug metabolism parameters were statistically analyze between the 2 groups.After routine general anesthesia induction(sufentanil 0.5 μg/kg),venous blood samples were collected to detect the changes in its concentration using high performance liquid chromatography-mass spectrometry(HPLC-MS).The pharmacokinetic data of sufentanil were calculated between the normal BMI group and overweight group in all participants and between the 2 body weight groups among those with different genotype combinations.Results Among the 90 participants completing the blood drug concentration test,8 patients had their blood samples contaminated(including 1 case with an anesthesia duration of<2 h),and 3 were excluded due to low weight or overweight.Eventually,79 participants were included in the pharmacokinetic analysis on the normal body weight group and the overweight group.Compared with the normal body weight group,the central compartment volume of distribution in the overweight group was significantly reduced(P<0.05),while no obvious differences were observed between the 2 groups in terms of peripheral compartment volume of distribution,total clearance rate,peripheral compartment clearance rate,distribution half-life,clearance half-life,and area under the blood concentration-time curve.In group Ⅰ(n=26),the overweight patients(n=13)had significantly reduced central compartment volume of distribution,peripheral compartment volume of distribution,and peripheral compartment clearance rate when compared with the normal body weight patients(n=13)(P<0.05),while no differences were observed in other pharmacokinetic parameters.In groups Ⅱ(n=25)and Ⅲ(n=28),the overweight patients and normal body weight patients had no statistical differences in all pharmacokinetic parameters.Conclusion Among the patients with the same genotype combination of CYP3A4/5 mutations,there was no difference in the metabolism of sufentanil between the overweight and normal weight patients.Additionally,in the population of 3A4 homozygous mutation or 3A4 heterozygous mutation+3A5 homozygous mutation,the overweight patients have smaller peripheral distribution range of sufentanil,and weakened metabolic process.