Alzheimer’s disease (AD) is a chronic neurodegenerative disorder that severely affects the health of the elderly, marked by its incurability, high prevalence, and extended latency period. The current approach to AD prevention and treatment emphasizes early detection and intervention, particularly during the pre-AD stage of mild cognitive impairment (MCI), which provides an optimal “window of opportunity” for intervention. Clinical detection methods for MCI, such as cerebrospinal fluid monitoring, genetic testing, and imaging diagnostics, are invasive and costly, limiting their broad clinical application. Speech, as a vital cognitive output, offers a new perspective and tool for computer-assisted analysis and screening of cognitive decline. This is because elderly individuals with cognitive decline exhibit distinct characteristics in semantic and audio information, such as reduced lexical richness, decreased speech coherence and conciseness, and declines in speech rate, voice rhythm, and hesitation rates. The objective presence of these semantic and audio characteristics lays the groundwork for computer-based screening of cognitive decline. Speech information is primarily sourced from databases or collected through tasks involving spontaneous speech, semantic fluency, and reading, followed by analysis using computer models. Spontaneous language tasks include dialogues/interviews, event descriptions, narrative recall, and picture descriptions. Semantic fluency tasks assess controlled retrieval of vocabulary items, requiring participants to extract information at the word level during lexical search. Reading tasks involve participants reading a passage aloud. Summarizing past research, the speech characteristics of the elderly can be divided into two major categories: semantic information and audio information. Semantic information focuses on the meaning of speech across different tasks, highlighting differences in vocabulary and text content in cognitive impairment. Overall, discourse pragmatic disorders in AD can be studied along three dimensions: cohesion, coherence, and conciseness. Cohesion mainly examines the use of vocabulary by participants, with a reduction in the use of nouns, pronouns, verbs, and adjectives in AD patients. Coherence assesses the ability of participants to maintain topics, with a decrease in the number of subordinate clauses in AD patients. Conciseness evaluates the information density of participants, with AD patients producing shorter texts with less information compared to normal elderly individuals. Audio information focuses on acoustic features that are difficult for the human ear to detect. There is a significant degradation in temporal parameters in the later stages of cognitive impairment; AD patients require more time to read the same paragraph, have longer vocalization times, and produce more pauses or silent parts in their spontaneous speech signals compared to normal individuals. Researchers have extracted audio and speech features, developing independent systems for each set of features, achieving an accuracy rate of 82% for both, which increases to 86% when both types of features are combined, demonstrating the advantage of integrating audio and speech information. Currently, deep learning and machine learning are the main methods used for information analysis. The overall diagnostic accuracy rate for AD exceeds 80%, and the diagnostic accuracy rate for MCI also exceeds 80%, indicating significant potential. Deep learning techniques require substantial data support, necessitating future expansion of database scale and continuous algorithm upgrades to transition from laboratory research to practical product implementation.

[摘 要] 目的：探究连翘苷（PHN）调节高迁移率族蛋白B1（HMGB1）/晚期糖基化终产物受体（RAGE）信号通路对胶质瘤U251细胞增殖、侵袭及上皮间质转化（EMT）的影响。方法：将人胶质瘤U251细胞分为PHN-0组（0 µmol/L PHN处理）、PHN低、中和高剂量组（PHN-50、PHN-100、PHN-200组，分别用50、100和200 µmol/L PHN处理）、PHN + pcDNA-NC组（转染pcDNA-NC质粒后200 µmol/L PHN处理）和PHN + HMGB1组（转染过表达HMGB1质粒后200 µmol/L PHN处理）。CCK-8法和克隆形成实验检测各组细胞的增殖能力，流式细胞术检测各组细胞的凋亡水平，Transwell实验检测各组细胞的迁移和侵袭能力，ELISA检测各组细胞分泌IL-8水平，免疫荧光法检测各组细胞中神经钙黏素（N-cadherin）和上皮钙黏素（E-cadherin）阳性率，WB法检测各组细胞中Toll样受体4（TLR4）、核因子-κB（NF-κB）、HMGB1、RAGE、N-cadherin、E-cadherin、细胞周期蛋白D1（cyclin D1）、细胞周期蛋白依赖性激酶2（CDK2）、B淋巴细胞瘤-2（Bcl-2）、Bcl-2相关X蛋白（BAX）蛋白的表达水平。结果：与PHN-0组相比，PHN-50、PHN-100、PHN-200组U251细胞增殖活力、克隆形成数、迁移和侵袭数、IL-8分泌水平、N-cadherin阳性率及其蛋白表达、TLR4、NF-κB、HMGB1、RAGE、cyclin D1、CDK2蛋白表达均显著降低（均P < 0.05），细胞凋亡率、E-cadherin阳性率及其蛋白表达、BAX/Bcl-2比值均显著升高（均P < 0.05）；同时过表达HMGB1则可逆转PHN对U251细胞增殖、迁移、侵袭及EMT的抑制作用和对凋亡的促进作用（均P < 0.05）。结论：PHN通过HMGB1/RAGE信号通路抑制胶质瘤U251细胞增殖、侵袭及EMT进程。

OBJECTIVES: To investigate the effect of ecliptasaponin A (ESA) for alleviating dextran sulfate sodium (DSS)-induced inflammatory bowel disease (IBD) in mice and the underlying mechanism. METHODS: Twenty-four male C57BL/6 mice (8-10 weeks old) were equally randomized into control group, DSS-induced IBD model group, and DSS+ESA (50 mg/kg) treatment group. Disease activity index (DAI), colon length and spleen index of the mice were measured, and intestinal pathology was examined with HE staining. The expressions of inflammatory mediators (TNF-α, IL-6, and iNOS) in the colon mucosa were detected using ELISA and RT-qPCR, and intestinal barrier integrity was assessed using AB-PAS staining and by detecting ZO-1 and claudin-1 expressions using immunofluorescence staining and Western blotting. In cultured RAW264.7 macrophages, the effects of treatment with 50 μmol/L ESA, alone or in combination with 20 μmol/L RO8191 (a JAK2/STAT3 pathway activator), on M1 polarization of the cells induced by LPS and IFN-γ stimulation and expressions of JAK2/STAT3 pathway proteins were analyzed using flow cytometry and Western blotting. RESULTS: In the mouse models of DSS-induced IBD, ESA treatment significantly alleviated body weight loss and colon shortening, reduced DAI, spleen index and histological scores, and ameliorated inflammatory cell infiltration in the colon tissue. ESA treatment also suppressed TNF‑α, IL-6 and iNOS expressions, protected the goblet cells and the integrity of the mucus and mechanical barriers, and upregulated the expressions of ZO-1 and claudin-1. ESA treatment obviously decreased CD86⁺ M1 polarization in the mesenteric lymph nodes of IBD mice and in LPS and IFN-γ-induced RAW264.7 cells, and significantly reduced p-JAK2 and p-STAT3 expressions in both the mouse models and RAW264.7 cells. Treatment with RO8191 caused reactivation of JAK2/STAT3 and strongly attenuated the inhibitory effect of ESA on CD86⁺ polarization in RAW264.7 cells. CONCLUSIONS ESA alleviates DSS-induced colitis in mice by suppressing JAK2/STAT3-mediated M1 macrophage polarization and mitigating inflammation-driven intestinal barrier damage.
Animals ; Mice ; Janus Kinase 2/metabolism* ; STAT3 Transcription Factor/metabolism* ; Mice, Inbred C57BL ; Male ; Dextran Sulfate ; Macrophages/cytology* ; Colitis/metabolism* ; Saponins/pharmacology* ; Signal Transduction/drug effects* ; RAW 264.7 Cells ; Triterpenes/pharmacology* ; Interleukin-6/metabolism*

OBJECTIVES: To investigate the mechanism through which pinostrobin (PSB) alleviates dextran sulfate sodium (DSS)-induced colitis in mice. METHODS: C57BL/6 mice were randomized into control group, DSS model group, and PSB intervention (30, 60, and 120 mg/kg) groups. Colitis severity of the mice was assessed by examining body weight changes, disease activity index (DAI), colon length, and histopathology. The expressions of tight junction proteins ZO-1 and claudin-1 in the colon tissues were examined using immunofluorescence staining, and macrophage infiltration and polarization were analyzed with flow cytometry. ELISA and RT-qPCR were used for detecting the expressions of inflammatory factors (TNF‑α and IL-6) and chemokines (CCL₂, CXCL₁₀, and CX₃CL₁) in the colon tissues, and PI3K/AKT phosphorylation levels were analyzed with Western blotting. In cultured Caco-2 and RAW264.7 cells, the effect of PSB on CCL₂-mediated macrophage migration was assessed using Transwell assay. Network pharmacology analysis was performed to predict the key pathways that mediate the therapeutic effect of PSB. RESULTS: In DSS-induced mouse models, PSB at 60 mg/kg optimally alleviated colitis, shown by reduced weight loss and DAI scores and increased colon length. PSB treatment significantly upregulated ZO-1 and claudin-1 expressions in the colon tissues, inhibited colonic macrophage infiltration, and promoted the shift of macrophage polarization from M1 to M2 type. In cultured intestinal epithelial cells, PSB significantly inhibited PI3K/AKT phosphorylation and suppressed chemokine CCL₂ expression. PSB treatment obviously blocked CCL₂-mediated macrophage migration of RAW264.7 cells, which could be reversed by exogenous CCL₂. Network pharmacology analysis and rescue experiments confirmed PI3K/AKT and CCL₂ signaling as the core targets of PSB. CONCLUSIONS PSB alleviates DSS-induced colitis in mice by targeting intestinal epithelial PI3K/AKT signaling, reducing CCL₂ secretion, and blocking macrophage chemotaxis and migration, highlighting the potential of PSB as a novel natural compound for treatment of inflammatory bowel disease.
Animals ; Mice ; Mice, Inbred C57BL ; Phosphatidylinositol 3-Kinases/metabolism* ; Colitis/drug therapy* ; Dextran Sulfate ; Proto-Oncogene Proteins c-akt/metabolism* ; Macrophages ; Chemokine CCL2/metabolism* ; Humans ; Signal Transduction/drug effects* ; Caco-2 Cells ; RAW 264.7 Cells ; Epithelial Cells/drug effects* ; Intestinal Mucosa/metabolism*

OBJECTIVE: To identify prognostic genes associated with lysosome-dependent cell death (LDCD) in patients with gastric cancer (GC). METHODS: Differentially expressed genes (DEGs) were identified using The Cancer Genome Atlas - Stomach Adenocarcinoma. Weighted gene co-expression network analysis was performed to identify the key module genes associated with LDCD score. Candidate genes were identified by DEGs and key module genes. Univariate Cox regression analysis, and least absolute shrinkage and selection operator regression and multivariate Cox regression analyses were performed for the selection of prognostic genes, and risk module was established. Subsequently, key cells were identified in the single-cell dataset (GSE183904), and prognostic gene expression was analyzed. Cell proliferation and migration were assessed using the Cell Counting Kit-8 assay and the wound healing assay. RESULTS: A total of 4,465 DEGs, 95 candidate genes, and 4 prognostic genes, including C19orf59, BATF2, TNFAIP2, and TNFSF18, were identified in the analysis. Receiver operating characteristic curves indicated the excellent predictive power of the risk model. Three key cell types (B cells, chief cells, and endothelial/pericyte cells) were identified in the GSE183904 dataset. C19orf59 and TNFAIP2 exhibited predominant expression in macrophage species, whereas TNFAIP2 evolved over time in endothelial/pericyte cells and chief cells. Functional experiments confirmed that interfering with C19orf59 inhibited proliferation and migration in GC cells. CONCLUSION C19orf59, BATF2, TNFAIP2, and TNFSF18 are prognostic genes associated with LDCD in GC. Furthermore, the risk model established in this study showed robust predictive power.
Stomach Neoplasms/pathology* ; Humans ; Prognosis ; Lysosomes/physiology* ; RNA-Seq ; Cell Death ; Single-Cell Analysis ; Gene Expression Regulation, Neoplastic ; Cell Proliferation ; Single-Cell Gene Expression Analysis

Pyran- and furanocoumarins are key representatives of tetrahydropyrans and tetrahydrofurans, respectively, exhibiting diverse physiological and medical bioactivities. However, the biosynthetic mechanisms for their core structures remain poorly understood. Here we combined multiomics analyses of biosynthetic enzymes in Peucedanum praeruptorum and in vitro functional verification and identified two types of key enzymes critical for pyran and furan ring biosynthesis in plants. These included three distinct P. praeruptorum prenyltransferases (PpPT1-3) responsible for the prenylation of the simple coumarin skeleton 7 into linear or angular precursors, and two novel CYP450 cyclases (PpDC and PpOC) crucial for the cyclization of the linear/angular precursors into either tetrahydropyran or tetrahydrofuran scaffolds. Biochemical analyses of cyclases indicated that acid/base-assisted epoxide ring opening contributed to the enzyme-catalyzed tetrahydropyran and tetrahydrofuran ring refactoring. The possible acid/base-assisted catalytic mechanisms of the identified cyclases were theoretically investigated and assessed using site-specific mutagenesis. We identified two possible acidic amino acids Glu303 in PpDC and Asp301 in PpOC as vital in the catalytic process. This study provides new enzymatic tools in the epoxide formation/epoxide-opening mediated cascade reaction and exemplifies how plants become chemically diverse in terms of enzyme function and catalytic process.

Objectives:To examine the proportion and trends of cervical adenocarcinoma in cervical cancer (ICC), mainly including cervical adenocarcinoma (CADC) and squamous cervical cancer (SCC) in China, and to analyze the distribution of human papilloma virus (HPV) in CADC and SCC.Methods:Published studies reporting HPVs distribution in various histological types or relative proportions of CADC in ICC in China were identified manually and searched systematically in Medline, Embase, Cochrane Library databases, CNKI and Wanfang since the databases were established until October 2022. Meta-analysis was performed using Stata 16.0 software. And we applied the random-effects models to estimate the combined effect values due to the high heterogeneity.Results:Twenty-three studies were eligible. The relative prevalence of CADC was 9.0% (95% CI, 7.7%－10.3%). According to the diagnosis time of ICC, the patients were divided into three time periods, which is 1979-2005, 2006-2011, 2012-2022 respectively. The prevalence of CADC by time was: 6.0% in 1979-2005, 8.1% in 2006-2011, and 9.5% in 2012-2022, respectively, with no statistically significant trend in proportions over time (χ 2=5.03, P=0.081). Meanwhile, the percentage of CADC also varies by regions, and the highest percentage of CADC was found in the eastern region (11.2%), followed by the western region (7.3%) and the central region (5.9%). The total prevalence of HPV infection in CADC was 72.3%, which was lower than 92.0% in SCC, and the difference was statistically significant (χ 2=300.89, P＜0.01). To be specific, the top three HPV types prevalent in CADC were HPV18 (45.0%), HPV16 (22.0%), and HPV52 (7.3%), and those prevalent in the SCC were HPV16 (64.2%), HPV52 (5.6%), HPV18 (5.4%). The results of the Egger's test, and Begg's test showed that there was no publication bias in this study and sensitivity analysis showed that the results of this study were fairly stable. Conclusions:The proportion of CADC in China has increased in a limited way in the past decades, and there are regional differences in the proportion of CADC. The predominant type is HPV18 in CADC and HPV16 in SCC. To eliminate the limitations of the secondary literature, a multicenter study with consistent diagnostic levels and identical HPV genotyping tests is still needed in the future to better characterize the relative proportion of cervical adenocarcinoma and the trend of HPV changes, which will provide a basis for the improvement of HPV vaccine and screening policies.

Objective:To report the results of national surveillance on the distribution and antimicrobial resistance profile of clinical Gram-negative bacteria isolates from bloodstream infections in China in 2022.Methods:The clinical isolates of Gram-negative bacteria from blood cultures in member hospitals of national bloodstream infection Bacterial Resistant Investigation Collaborative System（BRICS）were collected during January 2022 to December 2022. Antibiotic susceptibility tests were conducted by agar dilution or broth dilution methods recommended by Clinical and Laboratory Standards Institute（CLSI）. WHONET 5.6 and SPSS 25.0 software were used to analyze the data.Results:During the study period，9 035 strains of Gram-negative bacteria were collected from 51 hospitals，of which 7 895（87.4%）were Enterobacteriaceae and 1 140（12.6%）were non-fermenting bacteria. The top 5 bacterial species were Escherichia coli（ n=4 510，49.9%）， Klebsiella pneumoniae（ n=2 340，25.9%）， Pseudomonas aeruginosa（ n=534，5.9%）， Acinetobacter baumannii complex（ n=405，4.5%）and Enterobacter cloacae（ n=327，3.6%）. The ESBLs-producing rates in Escherichia coli， Klebsiella pneumoniae and Proteus spp. were 47.1%（2 095/4 452），21.0%（427/2 033）and 41.1%（58/141），respectively. The prevalence of carbapenem-resistant Escherichia coli（CREC）and carbapenem-resistant Klebsiella pneumoniae（CRKP）were 1.3%（58/4 510）and 13.1%（307/2 340）；62.1%（36/58）and 9.8%（30/307）of CREC and CRKP were resistant to ceftazidime/avibactam combination，respectively. The prevalence of carbapenem-resistant Acinetobacter baumannii（CRAB）complex was 59.5%（241/405），while less than 5% of Acinetobacter baumannii complex was resistant to tigecycline and polymyxin B. The prevalence of carbapenem-resistant Pseudomonas aeruginosa（CRPA）was 18.4%（98/534）. There were differences in the composition ratio of Gram-negative bacteria in bloodstream infections and the prevalence of main Gram-negative bacteria resistance among different regions，with statistically significant differences in the prevalence of CRKP and CRPA（ χ2=20.489 and 20.252， P<0.001）. The prevalence of CREC，CRKP，CRPA，CRAB，ESBLs-producing Escherichia coli and Klebsiella pneumoniae were higher in provinicial hospitals than those in municipal hospitals（ χ2=11.953，81.183，10.404，5.915，12.415 and 6.459， P<0.01 or <0.05），while the prevalence of CRPA was higher in economically developed regions（per capita GDP ≥ 92 059 Yuan）than that in economically less-developed regions（per capita GDP <92 059 Yuan）（ χ2=6.240， P=0.012）. Conclusions:The proportion of Gram-negative bacteria in bloodstream infections shows an increasing trend，and Escherichia coli is ranked in the top，while the trend of CRKP decreases continuously with time. Decreasing trends are noted in ESBLs-producing Escherichia coli and Klebsiella pneumoniae. Low prevalence of carbapenem resistance in Escherichia coli and high prevalence in CRAB complex have been observed. The composition ratio and antibacterial spectrum of bloodstream infections in different regions of China are slightly different，and the proportion of main drug resistant bacteria in provincial hospitals is higher than those in municipal hospitals.

Objective:To investigate the predictive value of preoperative D-dimer level for futile recanalization (FR) after mechanical thrombectomy (MT) in patients with acute ischemic stroke (AIS).Methods:It was a nested case-control study. A total of 116 patients with large vessel occlusion (LVO) stroke, who underwent successful recanalization (modified Thrombolysis in Cerebral Infarction, mTICI≥2b) after MT at the Stroke Unit of Beijing Hospital from August 2018 to January 2022,were consecutively enrolled, including 72 males (62.1%) with the age of (72.8±13.1) years. According to the 3-month modified Rankin Scale (mRS) score after MT, patients were divided into the meaningful recanalization group (mRS 0-2, n=41) and the futile recanalization group (mRS 3-6, n=75). The baseline clinical data of enrolled patients was collected. Logistic regression analysis was used to identify the independent risk factors for FR after MT in patients with AIS. Receiver operating characteristic (ROC) curve was used to evaluate the predictive value of D-dimer for FR. Results:Multivariate logistic regression analysis showed that high baseline systolic blood pressure (SBP) ( OR=1.038, 95% CI: 1.012-1.065, P=0.004), baseline National Institutes of Health Stroke Scale (NIHSS) score≥12 ( OR=10.157, 95% CI: 3.624-28.470, P<0.001) and high preoperative D-dimer level ( OR=4.536, 95% CI: 1.379-14.922, P=0.013) were independent predictors of FR after MT in AIS patients with LVO. ROC curve analysis indicated a good predictive value of preoperative D-dimer for the occurrence of FR ( AUC=0.733, 95% CI: 0.638-0.829, P<0.05), the optimal cut-off value of D-dimer was 2.65 μg/L(Lg), with the Youden index, sensitivity, specificity and accuracy of 0.435, 53.3%, 90.2% and 66.4%, respectively. Conclusion:High preoperative D-dimer level is an independent predictor of futile recanalization after MT in AIS patients with LVO, which shows good predictive ability for futile recanalization.