OBJECTIVE To investigate the improvement effects and mechanism of astragaloside Ⅳ （AS- Ⅳ ） on lipopolysaccharide （LPS）-induced neuroinflammation. METHODS BV2 cells were divided into control group， LPS group， AS-Ⅳ groups at concentrations of 20 and 40 μmol/L， and dexamethasone group （2 μmol/L）. Except for control group， neuroinflammation model was established with LPS （1 μg/mL） in other groups after medication. The levels of inflammatory factors [interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， and nitric oxide （NO）] in cell supernatant were measured in each group. Mice were randomly divided into normal group， model group， positive control group （Aspirin enteric-coated tablet， 20 mg/kg）， AS-Ⅳ low- and high-dose groups （10， 20 mg/kg）， with 6 mice in each group. Mice in each group were administered the corresponding drug/normal saline via gavage/intraperitoneal injection， once a day， for 14 consecutive days. Except for normal group， other groups were intraperitoneally injected with LPS （250 μg/kg） 1 hour after daily administration of the drug/normal saline to establish neuroinflammation model. Serum levels of IL-6 and TNF-α were measured 2 h after the last medication； histopathological morphology of cerebral tissue in mice were observed； the co-localization of inducible nitric oxide synthase （iNOS）/ionized calcium binding adapter molecule 1 （Iba1） and CD206/Iba1 in the cerebral cortex region of mice was observed； the expressions of proteins related to the nuclear factor-κB （NF-κB）/mitogen-activated protein kinase （MAPK） signaling pathway in brain tissue of mice were also determined， including NF-κB p65， phosphorylated NF-κB p65（p-NF-κB p65）， p38 MAPK， phosphorylated p38 MAPK （p-p38 MAPK）， extracellular signal-regulated kinase （ERK）， and phosphorylated ERK （p-ERK）. RESULTS In the cell experiments， compared with control group， the levels of IL-6， TNF- α and NO in the cell supernatant of the LPS group were increased significantly （P＜0.05）； compared with LPS group， the levels of IL-6， TNF-α and NO were decreased significantly in the administration groups （P＜0.05）. In the animal experiments， compared with the normal group， the serum levels of IL-6 and TNF- α， the number of iNOS/Iba1 co-localization positive cells in the cerebral cortex， and the phosphorylation levels of p38 MAPK， NF- κB p65 and ERK proteins in brain tissue were all significantly increased/elevated in model group （P＜0.05）； the number of CD206/ Iba1 co-localization positive cells in the cerebral cortex region significantly decreased （P＜0.05）. The neurons in the cerebral cortex and the CA3 region of the hippocampus displayed a disordered arrangement. Compared with model group， above quantitative indexes of mice were all reversed significantly in administration groups （P＜0.05）； the neuronal cells in the cerebral cortex and the CA3 region of the hippocampus exhibited a relatively orderly arrangement. CONCLUSIONS AS-Ⅳ may inhibit the activation of the NF-κB/MAPK signaling pathway， promote the M2-type polarization of microglia， and thereby suppress neuroinflammatory responses.

ObjectiveTo characterize the quality differences among different germplasm and introduced varieties of Bupleurum scorzonerifolium roots（BSR）， and explore the underlying molecular mechanisms， providing a basis for high-quality production and quality control. MethodsWild BSR from Yulin（YLW） served as the quality reference， we conducted comparative analysis among YLW， locally domesticated wild germplasm in Yulin（YLC3）， Daqing germplasm introduced and cultivated in Yulin（YLDQC3）， and locally cultivated germplasm in Daqing（DQC3）. A combination of traditional pharmacognostic methods and modern multi-omics analyses was employed， including macroscopic traits（appearance， odor）， microscopic features（proportions of cork， phloem， xylem）， cell wall component contents（hemicellulose， cellulose， lignin）， carbohydrate contents（starch， water-soluble polysaccharides）， marker compound contents（ethanol-soluble extracts， total saponins， liposoluble extracts， and saikosaponins A， B₂， C， D）， metabolomics， and transcriptomics， in order to systematically characterize quality differences and investigate molecular mechanisms among these samples. ResultsMacroscopically， Yulin-produced BSR（YLW， YLC3， YLDQC3） exhibited significantly greater weight， length， and upper and middle diameters than Daqing-produced BSR（DQC3）. Odor-wise， YLW and YLC3 had a a fragrance taste， YLDQC3 had a rancid oil odor， and DQC3 had a sweet and fragrant taste. Microscopically， Yulin germplasm（YLW， YLC3） and Daqing germplasm（YLDQC3， DQC3） shared similar structural features， respectively. However， Yulin germplasm showed significantly higher proportions of cork and phloem， as well as stronger xylem vessel staining intensity compared to Daqing germplasm. Regarding various component contents， Yulin germplasm contained significantly higher levels of ethanol-soluble extracts， total saponins， and saikosaponins A， B₂， C， D， while Daqing germplasm had significantly higher levels of hemicellulose， starch， and liposoluble extracts. After introduction to Yulin， the Daqing germplasm（YLDQC3） showed increased starch， water-soluble polysaccharides and liposoluble extracts contents， decreased cell wall component content， but no significant difference in other component contents. Metabolomics revealed that saponins and terpenes accumulated significantly in Yulin germplasm， while alcohols and aldehydes accumulated predominantly in Daqing germplasm. Transcriptomics indicated similar gene expression patterns within the same germplasm but specificity between different germplasms. Integrative metabolomic-transcriptomic analysis identified 145 potential key genes associated with the saikosaponin biosynthesis pathway， including one acetyl-coenzyme A（CoA） acetyltransferase gene（ACAT）， one 3-hydroxy-3-methylglutaryl-coenzyme A synthase gene（HMGS）， two hydroxymethylglutaryl-CoA（HMG-CoA） reductase genes（HMG）， one phosphomevalonate kinase gene（PMK）， one 1-deoxy-D-xylose-5-phosphate synthase gene（CLA）， one hydroxymethylbuten-1-aldol synthase gene（HDR）， two farnesyl pyrophosphate synthase genes（FPPS）， one squalene synthase gene（SQS）， one β-amyrin synthase gene（BAS）， 102 cytochrome P450（CYP450） gene family members， and 32 uridine diphosphate-glucuronosyltransferase（UGT） gene family members. ConclusionAmong the three cultivated types， YLC3 most closely resembles YLW in appearance， microscopic features， contents of major bioactive constituents， metabolomic and transcriptomic profiles. Yulin germplasm exhibits superior saponin synthesis capability compared to Daqing germplasm， and Yulin region is more suitable for the growth of B. scorzonerifolium. Based on these findings， it is recommended that artificial cultivation in northern Shaanxi and similar regions utilize the local Yulin germplasm source cultivated for at least three years.

Oral squamous cell carcinoma(OSCC)accounts for over 90％of oral malignancies,with more than 370 000 new cases and approximately 188 000 deaths annually worldwide.In China,there are roughly 65 000 new cases and 35 000 deaths each year,showing a significant upward trend compared with 2015 statistics.Despite continuous advancements in treatment modalities,the 5-year survival rate remains stagnant at 50％-60％,where tumor heterogeneity and therapy resistance persist as fundamental barriers to precision oncology.To address these critical challenges,this study established a standardized bioban-king protocol for OSCC patient-derived organoids(PDOs)(Patent:Method for constructing an oral squa-mous cell carcinoma organoid bank,ZL202311378598.3).Through groundbreaking optimization of cul-ture media,enzymatic digestion kinetics,and stepwise cryopreservation,we achieved a biobanking suc-cess rate exceeding 95％and pioneered synchronous cultivation of matched primary tumors,lymph node metastases,and adjacent normal mucosa from individual patients,preserving spatial heterogeneity and stromal interactions.Leveraging this platform,we developed high-throughput drug screening:Quantified heterogeneity-driven differential chemoresponse using adenosine triphosphate(ATP)-based viability as-says;We discovered resistance mechanisms:Identified sialylated cancer IgG(SIA-cIgG)-mediated cis-platin resistance(primary/secondary)through PTPN13 suppression,with anti-SIA-cIgG combination therapy demonstrating synergistic efficacy.Besides,we elucidated metastatic drivers:CRISPR-Cas9-edited organoids revealed WDR54 promoted metastasis via H3K4me3/H4K16ac epigenetic reprogramming,activating epithelial-mesenchymal plasticity(EMP)and inducing partial epithelial-mesenchymal transi-tion(pEMT).This"holographic patient-mirroring"platform provided unprecedented resolution for OSCC precision therapy and had been formally incorporated into the Chinese Stomatological Association Techni-cal Guidelines(Technical guideline for establishing patient-derived oral squamous cell carcinoma or-ganoid banks,CHSA 2024-08).Future integration of immune-competent organoids,3D-bioprinted vas-culature,and multi-omics-AI systems will accelerate personalized oncology.These innovations will accelerate clinical translation of personalized therapeutic regimens,ultimately bridging the gap between bench research and bedside application.

Objective To explore the effects and mechanism of calycosin-7-glucoside(CG)on lipopolysaccharide(LPS)-induced inflammatory injury in BV-2 cells and in a mouse model of neuroinflammation.Methods An in vitro neuroinflammation model was induced by LPS stimulation of BV-2 cells.BV-2 cells were divided into blank(CON),model(LPS),dexamethasone(DEX),and low-and high-dose CG(CG 10 μmol/L,CG 20 μmol/L,respectively)groups.The cell viability in each group was detected by Cell Counting Kit-8 assay,interleukin(IL)-6 and tumor necrosis factor(TNF)-α levels in the supernatant were detected by enzyme-linked immunosorbent assay(ELISA),and nitric oxide levels were detected using the Griess method.LPS was also used to induce neuroinflammation in mice in vivo.The mice were then divided randomly into blank(CON),model(LPS),aspirin,and low-and high-dose CG(CG 5 mg/kg,CG 10 mg/kg,respectively)groups.Pathological changes in the hippocampus were detected by hematoxylin/eosin staining.Serum levels of IL-6 and TNF-α were detected by ELISA,polarization of microglia was detected by immunofluorescence staining,and protein expression levels of Toll-like receptor 4(TLR4),myeloid differentiation primary response 88(MyD88),nuclear factor κB(NF-κB,P65)and phosphorylated-NF-κB(p-P65)in the cortex were detected by Western blot.Results CG alone or in combination with LPS in the concentration range of 2.5～160 μmol/L had no significant toxicity in BV-2 cells in vitro,compared with the CON group(P＞0.05).IL-6,TNF-α,and NO levels in the cell supernatant were increased in the LPS group compared with the CON group(P＜0.01),but were significantly reduced by CG(P＜0.05,P＜0.01).Hippocampal neurons were arranged loosely and disordered in the LPS group in vivo,compared with the CON group,and nuclear pyknosis was observed.Serum levels of IL-6 and TNF-α were increased(P＜0.05,P＜0.01).The number of ionized calcium binding adaptor molecule 1(Iba1)/inducible nitric oxide synthase(iNOS)cells was increased(P＜0.01),the number of CD206/Iba1 cells was decreased(P＜0.01),and expression levels of TLR4,MyD88,and p-P65 protein in the cortex were increased(P＜0.05).Compared with the LPS group,CG improved the pathological damage to the hippocampus and inhibited serum levels of IL-6 and TNF-α(P＜0.01).CG also decreased the number of iNOS/Iba1 cells,increased the number of CD206/Iba1 cells(P＜0.05,P＜0.01),and significantly down-regulated TLR4,MyD88,and p-P65 protein levels in the cortex(P＜0.05).Conclusions CG can ameliorate neuroinflammation in mice by suppressing the TLR4/MyD88/NF-κB pathway.

Objective To investigate the relationship between type 2 diabetes mellitus(T2DM)and cognitive decline.Methods Data were obtained from the cognitive impairment cohort of middle-aged and elderly population in rural areas of Xi'an City.The cohort consisted of residents aged 40 years and older in two villages of Huyi District,Xi'an.The baseline survey was completed between October 2014 and March 2015,with two follow-up visits in 2016 and 2018.The present study was conducted on cognitively normal people at baseline.Individual characteristics,lifestyle,and medical history were collected;physical and biochemical examinations were completed.According to medical history of T2DM and fasting blood glucose,the study population was divided into non-T2DM group,pre-existing T2DM group,and new-onset T2DM group.The Mini-Mental State Examination(MMSE)was used to assess global cognitive function.Participants with a drop of≥2 points in MMSE score from baseline after 4 years were defined as having cognitive decline.Chi-square test and multivariate Logistic regression analysis were employed to analyze the effect of T2DM status on the risk of cognitive decline.Results A total of 1 350 subjects completed the follow-up.In the follow-up population,1 096(81.2％)were free of T2DM,158(11.7％)already had T2DM at baseline,and 96(7.1％)developed new-onset T2DM during the follow-up.Cognitive decline was observed in 230 individuals after 4 years,representing 17.0％of the study population.The new-onset T2DM group had the highest 4-year incidence of cognitive decline(non-T2DM group vs.pre-existing T2DM group vs.new-onset T2DM group:15.7％vs.20.9％vs.26.0％,P=0.014),and the incidence of cognitive decline in the newly-onset T2DM group was significantly higher than that in the non-T2DM group(P=0.009).Multivariate Logistic regression analysis showed that the new-onset T2DM group had an increased risk of cognitive decline compared with the non-T2DM group within 4 years(OR=1.726,95％CI:1.029-2.896,P=0.039).However,no significant difference in 4-year risk of cognitive decline in the pre-existing T2DM group was observed(OR=1.402,95％CI:0.890-2.210,P=0.145).Conclusion Through the 4-year follow-up study of cognitively normal adults aged 40 and above in rural Xi'an,it was found that new-onset T2DM patients face a significantly elevated risk of cognitive decline,suggesting that cognitive decline may occur in the early stage of T2DM.

Objective:To investigate the role and mechanism of negative pressure wound therapy (NPWT) in a rabbit full-thickness wound model using non-targeted metabolomics.Methods:Eighteen male New Zealand rabbits (11-12 weeks old) were used. Two symmetrical circular full-thickness skin defects were created on the back of each rabbit. The animals were randomly divided into three groups: Control group (no treatment), Saline group (debridement with saline irrigation), and NPWT+ Saline group (saline debridement followed by 2 h of NPWT at -125 mm Hg once daily for two weeks). Wound healing was documented on days 0, 3, 7, 10, and 14. The wound healing rate was calculated as (original area-unhealed area)/original area × 100%. Histopathological changes were evaluated via hematoxylin and eosin (HE) staining. Metabolomic profiling of wound tissues was performed using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Differential metabolites were identified, and pathway enrichment analysis was conducted. Oxidative stress markers, including superoxide dismutase (SOD) and catalase (CAT) activities and malondialdehyde (MDA) content, were measured using commercial kits. Data were analyzed using SPSS 20.0. One-way ANOVA with Tukey’s HSD test or Welch’s ANOVA with Games-Howell test was applied as appropriate.Results:On days 3, 10, and 14, the wound healing rate in the NPWT+ Saline group was significantly higher than that in the Control and Saline groups ( P<0.05). On day 7, the NPWT+ Saline group showed a significantly higher healing rate than the Saline group ( P<0.01), but no significant difference compared with the Control group ( P>0.05). HE staining on day 7 revealed enhanced epithelialization, thicker granulation tissue, higher microvessel density, and more abundant, well-organized collagen in the NPWT+ Saline group. By day 14, all groups had formed relatively continuous epithelial structures. Non-targeted metabolomics identified riboflavin and spermidine as differential metabolites. Pathway analysis highlighted riboflavin metabolism and glutathione metabolism as the most significantly enriched pathways. Compared with the Control and Saline groups, the NPWT+ Saline group exhibited significantly increased CAT and SOD activities ( P<0.05) and decreased MDA content ( P<0.01), indicating reduced oxidative stress. Conclusion:NPWT may promote wound healing by elevating riboflavin and spermidine levels, thereby modulating riboflavin and glutathione metabolism and regulating local redox reactions.

Purpose To investigate the incidence and risk of malignancy(ROM)of the Bethesda class Ⅰ/Ⅲ di-agnosis of thyroid nodules due to insufficient number of follicular cells,and to analyze the correlation between their in-sufficient cell volume and the characteristics of the nodules themselves from the perspective of ultrasound and histology.Methods Clinical data were collected from fine needle aspiration cytology(FNAC)of the thyroid gland.Review and statistical analysis was performed on cases with the Bethesda class Ⅰ/Ⅲ diagnosis due to insufficient cell volume.The incidence and the ROM of Bethesda class Ⅰ/Ⅲ diagnosis were calculated.BRAF V600E(+)or postoperative patho-logical indicating papillary thyroid carcinoma(PTC)was used as the criterion for malignancy.Then,we matched the Bethesda class Ⅱ/Ⅵ cases with sufficient cell volume as the control group.The ultrasound characteristics and histo-logical images of the two groups were compared and analyzed in order to reveal the correlation between the insufficient amount of penetrating cells and the objective characteristics of the nodule itself.Results There were 39 solid thyroid nodules with the Bethesda class Ⅰ diagnosis,with an incidence of 3.3％and a ROM of 38.5％,and 160 nodules with the Bethesda class Ⅲ diagnosis,with an incidence of 13.5％and a ROM of 59.4％.The incidence and ROM of nod-ules with C-TIRADS ≥4b(22.4％,67.6％)were higher than those of C-TIRADS ≤4a(12.7％,39.8％),and the differences were statistically significant(P＜0.001).Compared to the Bethesda class Ⅱ/Ⅵ nodules with sufficient cell volume,occurrence of the Bethesda class Ⅰ/Ⅲ nodules were significantly correlated with small nodules(maximal diameter＜5 mm),vertical growth(aspect ratio ≥ 1)and poor blood supply(no or little blood flow signals)(r=0.131,-0.230,0.237,P=0.008,＜0.001,＜0.001).They were also significantly correlated with the pathologic histologic structure of diffuse significant fibrosis of the interstitium and low parenchyma/interstitium composition ratio(about 1:1)(r=-0.269,-0.396,P=0.019,＜0.001).Conclusion Thyroid Bethesda class Ⅰ/Ⅲ nodules have a high ROM,and BRAF V600E detection is recommended as a tool of tiered management.Bethesda class Ⅰ/Ⅲ diagnosis of insufficient cell volume is more likely when the nodules are too small,grow vertically and lack blood sup-ply,presumably associated with extensive interstitial fibrosis and sparse parenchymal cells.

Objective:To investigate the role and mechanism of negative pressure wound therapy (NPWT) in a rabbit full-thickness wound model using non-targeted metabolomics.Methods:Eighteen male New Zealand rabbits (11-12 weeks old) were used. Two symmetrical circular full-thickness skin defects were created on the back of each rabbit. The animals were randomly divided into three groups: Control group (no treatment), Saline group (debridement with saline irrigation), and NPWT+ Saline group (saline debridement followed by 2 h of NPWT at -125 mm Hg once daily for two weeks). Wound healing was documented on days 0, 3, 7, 10, and 14. The wound healing rate was calculated as (original area-unhealed area)/original area × 100%. Histopathological changes were evaluated via hematoxylin and eosin (HE) staining. Metabolomic profiling of wound tissues was performed using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Differential metabolites were identified, and pathway enrichment analysis was conducted. Oxidative stress markers, including superoxide dismutase (SOD) and catalase (CAT) activities and malondialdehyde (MDA) content, were measured using commercial kits. Data were analyzed using SPSS 20.0. One-way ANOVA with Tukey’s HSD test or Welch’s ANOVA with Games-Howell test was applied as appropriate.Results:On days 3, 10, and 14, the wound healing rate in the NPWT+ Saline group was significantly higher than that in the Control and Saline groups ( P<0.05). On day 7, the NPWT+ Saline group showed a significantly higher healing rate than the Saline group ( P<0.01), but no significant difference compared with the Control group ( P>0.05). HE staining on day 7 revealed enhanced epithelialization, thicker granulation tissue, higher microvessel density, and more abundant, well-organized collagen in the NPWT+ Saline group. By day 14, all groups had formed relatively continuous epithelial structures. Non-targeted metabolomics identified riboflavin and spermidine as differential metabolites. Pathway analysis highlighted riboflavin metabolism and glutathione metabolism as the most significantly enriched pathways. Compared with the Control and Saline groups, the NPWT+ Saline group exhibited significantly increased CAT and SOD activities ( P<0.05) and decreased MDA content ( P<0.01), indicating reduced oxidative stress. Conclusion:NPWT may promote wound healing by elevating riboflavin and spermidine levels, thereby modulating riboflavin and glutathione metabolism and regulating local redox reactions.

Objective To investigate the influencing factors associated with the risk of positive methylation levels of SHOX2/RASSF1A/PTGER4 genes in patients with pulmonary nodules with TCM susceptibility.Methods Using a cross-sectional study method,totally 165 patients with pulmonary nodules from the Respiratory Department of Wangjing Hospital,China Academy of Chinese Medical Sciences were included from May 2024 to December 2024.Clinical data such as patients'general medical history,nodule imaging reports,lung cancer gene methylation levels,and TCM constitution assessments were organized and statistically analyzed.On the basis of exploring the correlation between various variables and the risk of positive methylation of lung cancer genes through univariate correlation analysis,a binary logistic regression model was used for independent risk factor analysis.Results Univariate correlation analysis showed that age,gender,TCM susceptibility score,diameter of pulmonary nodules and upper lobe of the right lung were correlated with positive methylation levels of the SHOX2/RASSF1A/PTGER4 genes(P=0.03,P=0.02,P=0.01,P<0.01,P=0.03);and binary Logistic regression analysis showed that yang-deficiency quality,qi deficiency,qi depression and phlegm-dampness,the four pulmonary nodules susceptibility body scores were significantly and positively correlated with the risk of SHOX2/RASSF1A/PTGER4 gene methylation positivity.Compared with yang deficiency,phlegm-dampness and qi depression had a higher risk of gene methylation positivity for patients with pulmonary nodules(P=0.04,P=0.02).Conclusion Older age,males,and TCM susceptibility body correlated with the lung cancer gene methylation positive risk are correlated,in which phlegm dampness and qi depression are independent risk factors.

Objective To observe the application effect of surgical adhesive membrane combined with modified towel placement method in patients undergoing orthopedic unilateral biportal endoscopy(UBE)operation.Methods A total of 61 patients who underwent orthopedic UBE operation in our hospital from December 2022 to September 2023 were selected and divided into the control group(31 cases,conventional surgical adhesive membrane and towel placement)and the observation group(30 cases,surgical adhesive membrane combined with modified towel placement method)according to different placement methods of membrane and towel during operation.The degree of wetting in the surgical area,vital signs,intraoperative adverse events,postoperative incision infection rate and patients'satisfaction were compared between the two groups.Results The proportions of wet sterile sheets around the surgical area and moist skin of patients in the observation group were lower than those in the control group(P<0.05).The saturation of peripheral oxygen(SpO2),heart rate(HR),mean arterial pressure(MAP)at spinal canal decompression and the end of the operation of patients in the observation group were significantly better than those in the control group(P<0.05).No postoperative incision infection occurred in the two groups,and there was no statistically significant difference in the total incidence of adverse events between the two groups(P>0.05).The patients'satisfaction in the observation group was significantly higher than that in the control group(P<0.05).Conclusion Surgical adhesive membrane combined with modified towel placement method can reduce the degree of wetting in the surgical area during orthopedic UBE operation,stabilize the patients'vital signs,and improve patients'satisfaction.