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.

Viral pneumonia in the Lancang-Mekong River Basin exhibits distinct regional epidemiological characteristics and pathogenic mechanisms. Filthy dampness constitutes the core pathogenic factor of the disease here， and the core pathogenesis lies in the yang failing to be stored in the you （酉） position. Clinically， the fundamental therapeutic principle is established as boosting qi to expel filth and guiding yang qi to descend and converge into the you position. In the early stage， the pathogenesis is characterized by the pervasive diffusion of filthy dampness， yang qi floating externally and failing to be stored， and the lung failing to diffuse qi. The treatment focuses on invigorating yang qi to dispel filth and dredging the you pathways， primarily using modified Ganlu Xiaodu Decoction （甘露消毒丹）. In the progressive stage， the pathogenesis involves the tenacious stagnation of filthy dampness， yang constraint transforming into heat， and the rampant spread of heat-toxins. The therapeutic approach shifts to acrid medicinals for opening and bitter medicinals for promoting descent， rectifying healthy qi to expel filth， utilizing modified Dayuan Beverage （达原饮）. In the recovery stage， filthy dampness gradually recedes， accompanied by the depletion of qi and yin， and insufficient kidney yang. The treatment emphasizes supplementing qi and nourishing yin， accompanied by warming yang， with modified Yunv Decoction （玉女煎）.

Objective To explore and verify the prognostic value of endothelial cells in glioblastoma. Methods Through bioinformatics analysis of the TCGA and CGGA databases, we screened endothelial cell-related markers in GBM single-cell data according to a series of criteria. Moreover, univariate Cox regression analysis was performed to obtain and screen endothelial cell prognosis-related markers and construct endothelial cell-related prognostic risk score. qPCR experiments was used to verify the differences in the expression of prognostic markers in GBM tissues and peritumoral normal brain tissues. Kaplan-Meier method was used to construct the survival curve to identify the prognostic efficacy of the prognostic risk score. Results A total of 2 115 prognostic genes of glioblastoma (GBM) were screened. Among them, 1 494 was upregulated and 621 was downregulated. Seven groups of cells were obtained after GBM single-cell sequencing analysis, including AC-like tumor cells, endothelial cells, monocytes/macrophages, NB-like tumor cells, neurons, OC-like tumor cells, and OPC-like tumor cells. According to the differential genes of endothelial cells and the corresponding screening criteria, four genes (DUSP6, STC1, VWA1, and TM4SF1) were screened for risk-score construction. The expression of the target gene in GBM tissues and normal brain tissues around the tumor was significantly up-regulated detected by qPCR. The risk score=0.171*DUSP6+0.144*STC1+0.041*VWA1−0.004*TM4SF1. Conclusion The glioblastoma endothelial cells’ risk score determined in this study can preferably predict the prognosis of patients.

Objective: To construct a hypertension risk prediction model for residents aged 18-79 years, so as to provide an assessment tool for early screening and prevention of hypertension in high-risk groups. Methods: The permanent residents aged 18-79 years from 6 townships (streets) in Fangshan District of Beijing Municipality were selected as the study subjects using a multi-stage stratified random sampling method from March to June 2023. Demographic information, lifestyle, body mass index (BMI), blood pressure, fasting blood glucose, and blood lipid were collected through questionnaire survey, physical examination, and laboratory tests. Subjects were randomly divided into training and validation sets at a 7∶3 ratio. The logistic regression model was used to screen the risk factors of hypertension, and a hypertension risk prediction nomogram was constructed. Receiver operating characteristic (ROC) curve, calibration curve, and decision curve analysis were used to verify the discrimination, fit, and clinical application value of the model. Results: A total of 4 438 subjects were included, including 2 365 males (53.29%) and 2 073 females (46.71%), with a mean age of (44.99±14.90) years. The prevalence of hypertension was 35.29% (1 566 cases), and the standardized prevalence was 24.74%. The logistic regression model screened out 9 influencing factors of hypertension. The nomogram was established as ln[p/ (1-p)]= -2.873 + 0.935×40-<50 years + 1.463×50-<60 years + 1.908×60-<70 years + 2.346×70-79 years + 0.298×male-0.675×college degree or above + 0.384×smoking + 0.227×drinking + 0.572×overweight + 1.449×obesity + 0.557×heart rate ≥80 beats/min + 0.428×diabetes + 0.484×dyslipidemia. The area under the ROC curve of the validation set was 0.821 (95%CI: 0.798-0.843), and the calibration curve results showed that the calibration curve fitted the actual curve well. Decision curve analysis showed that the threshold probability was in the range of 0.10 to 0.70, and the model had good predictive value and clinical application value. Conclusion The nomogram based on age, gender, educational level, smoking, drinking, body mass index, heart rate, diabetes, and dyslipidemia can be used to predict the risk of hypertension among residents aged 18-79 years.

Objective To explore the application value of anhydrous ethanol as an alternative to methanol in the preparation of chromosomal specimens from peripheral blood lymphocytes, and to establish a set of quantitative analytical methods for objectively evaluating the effectiveness of specimen preparation. Methods Residual blood samples from routine laboratory slide preparation were used for lymphocyte culture. The standard slide preparation method was employed. The fixative in the control group was methanol and glacial acetic acid (3∶1). Four experimental groups were set up based on the ratio of anhydrous ethanol to glacial acetic acid in the fixative (volume ratios of 3∶1, 5∶1, 7∶1, and 9∶1 for experimental groups 1, 2, 3, and 4, respectively). A chromosomal analysis was conducted using an automated chromosome scanning/image analysis system to evaluate the morphology and dispersion of metaphase chromosomes in both control and experimental groups. Comparisons were made between the control and experimental groups regarding the dic + r aberration rate, ace aberration rate, chromosomal aberration rate, chromosome dispersion index, chromosome overlapping ratio, and dispersion index/overlapping ratio. Results Microscopic evaluation revealed that the preparation quality of experimental groups 1 and 2 was comparable to the control group. No statistically significant differences were observed in dic + r aberration rate between each of the experimental groups and the control (P > 0.05). All experimental groups except group 4 showed no significant differences in ace aberration rate and chromosome aberration rate compared with the control group (P > 0.05). Experimental groups 1 and 2 showed no significant differences in chromosome dispersion index, overlapping ratio, and dispersion index/overlapping ratio compared with the control group (P > 0.05). Conclusion A mixture of anhydrous ethanol and glacial acetic acid at a 5∶1 ratio is recommended for use as a fixative in the preparation of chromosomal specimens from peripheral blood lymphocytes. A quantitative index system for assessing the quality of chromosomal specimens was established, enabling objective evaluation of slide preparation effectiveness.

Ulcerative colitis (UC) is a chronic and non-specific inflammatory bowel disease (IBD). Huanglian Ganjiang decoction (HGD), derived from ancient book Beiji Qianjin Yao Fang, has demonstrated efficacy in treating UC patients traditionally. Previous research established that the compatibility of cold herb Coptidis Rhizoma + Phellodendri Chinensis Cortex (CP) and hot herb Angelicae Sinensis Radix + Zingiberis Rhizoma (AZ) in HGD synergistically improved colitis mice. This study investigated the compatibility mechanisms through which CP and AZ regulated inflammatory balance in colitis mice. The experimental colitis model was established by administering 3% dextran sulphate sodium (DSS) to mice for 7 days, followed by CP, AZ and CPAZ treatment for an additional 7 days. M1/M2 macrophage polarization levels, glucose metabolites levels and pyruvate dehydrogenase kinase 4 (PDK4) expression were analyzed using flow cytometry, Western blot, immunofluorescence and targeted glucose metabolomics. The findings indicated that CP inhibited M1 macrophage polarization, decreased inflammatory metabolites associated with tricarboxylic acid (TCA) cycle, and suppressed PDK4 expression and pyruvate dehydrogenase (PDH) (Ser-293) phosphorylation level. AZ enhanced M2 macrophage polarization, increased lactate axis metabolite lactate levels, and upregulated PDK4 expression and PDH (Ser-293) phosphorylation level. TCA cycle blocker AG-221 and adeno-associated virus (AAV)-PDK4 partially negated CP's inhibition of M1 macrophage polarization. Lactate axis antagonist oxamate and PDK4 inhibitor dichloroacetate (DCA) partially reduced AZ's activation of M2 macrophage polarization. In conclusion, the compatibility of CP and AZ synergistically alleviated colitis in mice through M1/M2 macrophage polarization balance via PDK4-mediated glucose metabolism reprogramming. Specifically, CP reduced M1 macrophage polarization by restoration of TCA cycle via PDK4 inhibition, while AZ increased M2 macrophage polarization through activation of PDK4/lactate axis.
Animals ; Drugs, Chinese Herbal/chemistry* ; Mice ; Macrophages/immunology* ; Glucose/metabolism* ; Pyruvate Dehydrogenase Acetyl-Transferring Kinase/genetics* ; Male ; Mice, Inbred C57BL ; Humans ; Colitis/drug therapy* ; Disease Models, Animal ; Colitis, Ulcerative/drug therapy* ; Metabolic Reprogramming

Objective To investigate the association between plasma homocysteine(Hcy)level and coronary artery calcification(CAC)and its clinical predictive efficacy.Methods A total of 172 patients who underwent coronary CT angiography North China University of Science and Technology Affiliated Hospital from April 2019 to May 2021 and CAC score(CACS)＞0 were enrolled.According to the CACS value,the subjects were divided into mild calcification group(n=136)and severe calcification group(n=36),and the clinical characteristics of two groups were compared and analyzed.Multivariate Logistic regression model was used to screen the independent influencing factors of CAC severity,and a prediction model was constructed based on the Hcy detection value.The clinical diagnostic value was evaluated by the receiver operating characteristic(ROC)curve.Results There were significant differences in Hcy,white blood cell count,triglyceride and magnesium ion levels between two groups(P＜0.05).Multivariate Logistic regression analysis showed that Hcy,white blood cell count and magnesium ion level were independent risk factors for the progression of CAC.Furthermore,a regression model based on Hcy was constructed and ROC curve was fitted to evaluate its predictive efficacy.The results suggested that the predictive model had the best performance when the critical value of Hcy was set at 27.4μmol/L:the sensitivity was 55.6％,the specificity was 97.1％,and the area under the curve was 0.765.Conclusion Hcy serves as an independent risk factor for the severity of CAC and can effectively predict the progression of CAC with high accuracy.

Objective:To identify and validate the key genes of ferroptosis in phospholipase A2 receptor (PLA2R) associated membranous nephropathy through bioinformatics analysis and in vitro experiments, and to explore the potential role of ferroptosis in PLA2R associated membranous nephropathy (PMN). Methods:The GSE115857 dataset obtained by retrieving the Gene Expression Omnibus (GEO) database and the ferroptosis-related genes obtained by retrieving the FerrDb database were intersected. The intersected genes were subjected to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The key ferroptosis genes associated with PMN were identified by intersecting genes selected using support vector machines-recursive feature elimination and least absolute shrinkage and selection operator regression. The results were validate by real-time PCR, cell counting kit-8, Western blotting and immunofluorescence in human renal podocyte line AB 8/13 from both the control group and model group.Results:A total of 25 genes related to ferroptosis of PMN were obtained, and GO and KEGG analysis showed that these genes were mainly involved in cell ferroptosis metabolism. The key ferroptosis genes of PMN obtained by machine learning method were activating transcription factor 3 ( ATF3) and coiled coil domain containing 6 ( CCDC6). The results of in vitro experiments showed that the human renal podocyte line AB 8/13 in the model group was significantly deformed and retracted compared with the control group. The surface area density of foot processes was significantly reduced, and the podocyte cytoskeleton was allosteric. The morphology of F-actin was disordered and the expression of synaptopodin was decreased. The cell proliferation activity was significantly decreased ( P<0.05). The expression of PLA2R protein was increased ( P<0.05), and the expression of GPX4 protein was decreased ( P<0.01). The protein and mRNA levels of ATF3 and CCDC6 were significantly up-regulated (all P<0.05). Conclusions:Ferroptosis may be one of the key mechanisms in the occurrence and development of PMN. In vitro experiments show that ATF3 and CCDC6 are the key genes in the ferroptosis of PMN podocytes, which provides new insights and ideas for the pathogenesis of PMN.

Background:Liver cirrhosis is characterized by chronic inflammation,progressive fibrosis,and eventual liver dysfunction,and poses a major global health challenge.Aims:To assess the global burden of liver cirrhosis and its risk factors from 1990 to 2021.Methods:Using data extracted from the Global Burden of Diseases,Injuries,and Risk Factors Study(GBD)2021,the incidence,mortality,disability-adjusted life years(DALYs),and their age-standardized rates of liver cirrhosis were analyzed.Furthermore,a stratified analysis was conducted by sex,age,region,and etiology,with projections of the trends in the next 15 years.Results:Compared to 1990,the global incidence number of liver cirrhosis in 2021 was increased by 58.2%,the death number and DALYs rose by 39.5%and 27.9%,respectively.While the global age-standardized incidence rate(ASIR)showed a slight increase,the age-standardized death rate(ASDR)and DALY rate continued to decline.Both ASIR and ASDR exhibited negative correlations with the sociodemographic index(SDI).All age-standardized rates were higher in males than in females.Since 1990,the incidence rate increased in younger populations,while the mortality and DALY rates declined in most age groups.Non-alcoholic fatty liver disease(NAFLD)emerged as the leading cause of incidence,whereas chronic hepatitis B and C remained the primary contributors to deaths and DALYs.The incidence of NAFLD was prominent in high and high-middle SDI regions,while chronic hepatitis B was concentrated in low SDI regions.Projections to 2036 indicated a continuing rise in ASIR,and declines in ASDR and DALY rate.The incidence of chronic hepatitis B was projected to decrease markedly,whereas that of NAFLD was expected to continue increasing.Conclusions:Between 1990 and 2021,the global incidence of liver cirrhosis showed a modest increase;in contrast,both mortality and DALY rates demonstrated a steady decline.Burden of liver cirrhosis poses notable regional disparities.NAFLD dominates incidence in high-income regions,while viral hepatitis remains predominant in low-income areas,highlighting the need for region-specific prevention strategies.

Objective By reviewing the access policies for new antimicrobial drugs internationally,this study provides a reference for the development of China's antimicrobial drug access policy.Methods The Chinese and English databases,offi-cial websites of health agencies,and health technology assessment agencies of various countries(CNKI,PubMed,NICE in the UK,HAS in France,etc.)were searched to collect and sort out the access policies of new antibacterial drugs.Taking the antibacte-rial drug ceftazidime-avibactam and the antifungal drug esaconazole sulfate as examples,the key evidence points for the evaluation of antibacterial drugs in the market access process of medical insurance were analyzed.Finally,the author put forward suggestions for China's antibacterial drug access policy based on the opinions of experts in related fields of antibacterial drugs in China.Re-sults Access policies for antimicrobial drugs had already established in the UK,US,EU,Sweden Switzerland,and Republic of Korea.These policies included a series of measures such as evaluation approval rewards,patent extensions,additional reimburse-ment,decoupling sales revenue from sales volume,etc.,with the ultimate goal being to encourage research and development of new antibiotics while reducing resistance rates.The review found that besides evaluating the safety,economy,and cost-effectiveness,some evaluation agencies also consider the actual clinical value and social value of drugs when reviewing the evidence of two types of new antibacterial drugs in the medical insurance access link.Conclusion China can draw upon international framework principles while considering unique demands such as national antibiotic management policy or national healthcare negotiation re-quirements to provide certain policy support throughout the market access process for new antimicrobials due to their unique val-ues so that it can encourage research & development while curbing antibiotic resistance.