ObjectiveTo investigate the effect of Yaoshu Zhuyu Fang on the regulation of the microRNA-17-5P (miR-17-5P)/murine double minute 2 (MDM2)/p53 axis in the proliferation and apoptosis of rat intervertebral disc annulus fibrosus cells, and its potential molecular mechanism. MethodsIntervertebral disc annulus fibrosus tissues were obtained from 8-week-old SPF-grade male SD rats, and annulus fibrosus cells were isolated and obtained by enzyme digestion and mechanical dispersion. Annulus fibrosus cells were divided into 6 groups: Group C was the blank control group, in which annulus fibrosus cells were not treated with interleukin-1β (IL-1β) but were cultured in RPMI 1640 complete medium. Group β was the degeneration model group constructed by treating annulus fibrosus cells with 10 ng/mL IL-1β for 24 h. Group β+B was the IL-1β + blank serum group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then treated with RPMI 1640 medium containing 5% blank serum for 24 h. Group β+W was the IL-1β + Yaoshu Zhuyu Fang-containing serum group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then treated with RPMI 1640 medium containing 5% Yaoshu Zhuyu Fang-containing serum for 24 h. Group β+I was the IL-1β + miR-17-5P inhibitor group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then transfected with miR-17-5P inhibitor. Group β+I+W was the IL-1β + miR-17-5P inhibitor + Yaoshu Zhuyu Fang-containing serum group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then transfected with miR-17-5P inhibitor, and finally treated with RPMI 1640 medium containing 5% Yaoshu Zhuyu Fang-containing serum for 24 h. CCK-8 assay was used to detect cell survival rate. Flow cytometry was used to detect cell apoptosis. Real-time quantitative PCR was used to detect the expression levels of miR-17-5P, MDM2 mRNA, and p53 mRNA in cells. Western blotting was used to detect the protein expression levels of MDM2 and p53 in cells. Dual-luciferase reporter system was used to analyze the targeting relationship between miR-17-5P and MDM2. ResultsCompared with Group C, Group β showed a significant decrease in cell survival rate (P<0.001), a significant increase in cell apoptosis rate (P<0.001), significantly increased expression of miR-17-5P, p53 mRNA, and p53 protein (P<0.001), and significantly decreased expression of MDM2 mRNA and protein (P<0.001). Compared with Group β, Group β+W, Group β+I, and Group β+I+W showed significantly increased cell survival rate, significantly decreased apoptosis rate, significantly decreased expression of miR-17-5P, p53 mRNA, and p53 protein, and significantly increased expression of MDM2 mRNA and protein (P<0.001). Moreover, changes in the above indicators were greater in Group β+I+W (P<0.001). Circular RNA Interactome predicted that miR-17-5P had specific binding sites with the 3' untranslated region (3'UTR) of MDM2. Transfection of miR-17-5P mimic significantly reduced the luciferase expression level of co-transfected luciferase reporter plasmid containing wild-type MDM2 3'UTR (P<0.05), but had no significant effect on luciferase expression in cells co-transfected with luciferase reporter plasmid containing mutant MDM2 3'UTR (P>0.05). ConclusionYaoshu Zhuyu Fang down-regulates the expression of miR-17-5P, promotes the synthesis of MDM2 protein, thereby down-regulates p53, promotes proliferation, and inhibits the apoptosis of rat intervertebral disc annulus fibrosus cells.

ObjectiveThis paper aims to investigate the potential molecular biological mechanism of Buyang Huanwu Tongfeng decoction in treating hyperuricemia and gouty arthritis by network pharmacology and molecular docking technology and preliminarily verify the mechanism through animal experiments. MethodsThe active ingredients and targets in the Buyang Huanwu Tongfeng decoction were obtained by the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and ETCM databases. The DisGeNET and GeneCards databases were utilized to acquire disease targets associated with hyperuricemia and gouty arthritis. These disease targets were then intersected with drug targets to identify key targets. The R language ClusterProfiler package and Python were employed for conducting gene ontology（GO） enrichment analysis and Kyoto encyclopedia of genes and genomes（KEGG） enrichment analysis. The regulatory network diagram of the drug-key target-function-pathway was visualized using Cytoscape 3.9.1 software， and the protein-protein interaction （PPI） network for key targets was depicted. Finally， the hub gene was determined through topological analysis. Auto Dock， PyMOL， and other software were used for molecular docking to explore the possible therapeutic mechanism of Buyang Huanwu Tongfeng decoction for hyperuricemia and gouty arthritis. In animal experiments， a composite rat model of hyperuricemia induced by intraperitoneal injection of oteracil potassium combined with gouty arthritis induced by the modified Coderre method was established. Through hematoxylin-eosin（HE） staining， uric acid test， enzyme linked immunosorbent assay（ELISA）， Western blot， and real-time polymerase chain reaction（Real-time PCR）， the molecular mechanism and key targets of Buyang Huanwu Tongfeng decoction for treating hyperuricemia and gouty arthritis were observed. ResultsAfter screening and removing duplicate values， 76 active ingredients and 15 key targets were finally obtained. GO enrichment analysis yielded that the treatment of hyperuricemia and gouty arthritis with Buyang Huanwu Tongfeng decoction was significantly associated with acute inflammatory response， astrocyte activation， regulation of interleukin （IL）-8 production， nuclear receptor activity， and binding of growth factor receptor. KEGG pathway enrichment analysis obtained that the key target genes were significantly associated with the IL-17 signaling pathway， advanced glycosylation end/receptor of advanced glycation endproducts（AGE/RAGE） signaling pathway， anti-inflammatory， and other pathways. PPI network indicated that albumin（ALB）， peroxisome proliferator-activated receptor-γ （PPAR-γ）， IL-6， IL-1β， and C-reactive protein（CRP） were the key protein targets. The molecular docking results showed that ALB had the strongest binding force with beta-carotene （β-carotene）. Biochemical results showed that blood uric acid decreased in the Buyang Huanwu Tongfeng decoction groups. HE staining results showed that the low-dose （7.76 g·kg^-1·d^-1）， medium-dose （15.53 g·kg^-1·d^-1）， and high-dose （31.05 g·kg^-1·d^-1） groups of Buyang Huanwu Tongfeng decoction had different degrees of remission， and the remission of the high-dose group was the most obvious. Fibroblastic tissue hyperplasia in synovial joints accompanied with inflammatory cell infiltration， as well as inflammatory cell infiltration in renal tissue of the high-dose group was significantly reduced， followed by the medium-dose and low-dose groups， and the expression of ALB， PPAR-γ， IL-6， IL-1β， and CRP was down-regulated to different degrees. ConclusionBy regulating the targets such as ALB， PPAR-γ， IL-6， IL-1β， and CRP， inhibiting the PPAR-γ/nuclear transcription factor （NF）-κB pathway， and reducing AGEs/RAGE-mediated inflammation， Buyang Huanwu Tongfeng decoction exerts anti-inflammatory and analgesic effects and activates blood circulation and diuresis in the treatment of hyperuricemia and gouty arthritis.

Background::B-cell maturation antigen (BCMA)-directed chimeric antigen receptor T (CAR-T) therapy yield remarkable responses in patients with relapsed/refractory multiple myeloma (R/RMM). Circulating tumor DNA (ctDNA) reportedly exhibits distinct advantages in addressing the challenges posed by tumor heterogeneity in the distribution and genetic variations in R/RMM.Methods::Herein, the ctDNA of 108 peripheral blood plasma samples from patients with R/RMM at the First Affiliated Hospital, School of Medicine, Zhejiang University was thoroughly investigated before administration of anti-BCMA CAR-T therapy to establish its predictive potential. Flow cytometry is used primarily to detect subgroups of T cells or CAR-T cells.Results::In this study, several tumor and T cell effector-mediated factors were considered to be related to treatment failure by an integrat analysis, including higher percentages of multiple myeloma (MM) cells in the bone marrow ( P = 0.0125), lower percentages of CAR-T cells in the peripheral blood at peak ( P = 0.0375), and higher percentages of CD8 + T cells ( P = 0.0340). Furthermore, there is a substantial correlation between high ctDNA level (>143 ng/mL) and shorter progression-free survival (PFS) ( P = 0.007). Multivariate Cox regression analysis showed that high levels of ctDNA (>143 ng/mL), MM-driven high-risk mutations (including IGLL5 [ P = 0.004], IRF4 [ P = 0.024], and CREBBP [ P = 0.041]), number of multisite mutations, and resistance-related mutation ( ERBB4, P = 0.040) were independent risk factors for PFS. Conclusion::Finally, a ctDNA-based risk model was built based on the above independent risk factors, which serves as an adjunct non-invasive measure of substantial tumor burden and a prognostic genetic feature that can assist in predicting the response to anti-BCMA CAR-T therapy.

Objective:To compare the types and contents of trace elements in compound trace element preparations used in parenteral nutrition in Beijing against the doses recommended by the guidelines, so as to facilitate optimal choice of preparations for different populations in clinical practice.Methods:The available compound trace element preparations were determined according to the drug list from the Medical Insurance Recruitment and Procurement System of Beijing and the recommended doses of trace elements in the guidelines released by Chinese domestic and foreign nutrition societies were collected and compared.Results:The compound trace element preparations used in Beijing comprised 4 varieties and 6 specifications, with the daily cost ranging from 11.32 to 471 Chinese Yuan. There were 3 varieties of trace element preparations for adults, including multi-trace elements injection (Ⅱ), multi-trace elements injection (Ⅲ), and multi-trace elements injection. Among those, multi-trace elements injection (Ⅲ) met the guideline recommendations best and the price was moderate. For pediatric populations, multi-trace elements injections (Ⅰ) satisfied better the trace element requirements for preterm infants and infants from term to 3 months, while multi-trace elements injections (Ⅲ) outperformed in meeting the trace element requirements for 3-to-12-month infants and children. However, there were still some differences in the trace element contents between the two preparations and the guideline recommendations.Conclusion:The currently available varieties and specifications of trace element preparations basically meet the recommendations of evidence-based guidelines, but there is still the need to improve the instructions for and evidence on the pediatric application of certain preparations.

Objective:An inceasing number of commercial multi-chambered bag products for parenteral nutrition (PN) have been launched in China, with some being included in the National Medical Insurance Catalogue (NMIC). In this study, the commercially available triple-chambered bag (3CB) products for PN in China were comprehensively analyzed in terms of basic characteristics, applicable population, and costs to guide the rational use of these products in clinical settings.Methods:The marketing information of 3CB products was collected. The inclusion of these products in NMIC and the Catalogue of Selected Drugs for National Centralized Procurement was investigated, along with the collection of manufacturer's instructions and pricing information in Beijing. Two researchers extracted and calculated the information of 3CB products, including energy density, types and levels of each component, glycolipid ratio, osmotic pressure, and price. The applicable population and economy of these products were also compared.Results:As of April 2024, 11 3CB products, including 23 specifications, had been approved in China. Eight products were included by NMIC and 2 by the Catalogue of Selected Drugs for National Centralized Procurement. The energy density, protein content, and lipid content dramatically differed among 3CB products (energy density, 254.0–493.7 kJ/100 ml; protein, 2.2–5.6 g/100ml; lipid, 2–4 g/100 ml; and glycolipid ratio, 62∶38–40∶60). Five products could only be infused via the central veins. The sources of fat emulsions included soybean oil-based long-chain fat-milk, medium-long-chain fat-milk, olive oil-based fat-milk, and fish oil-based fat-milk. All the 3CB products contained 8 essential amino acids and some non-essential amino acids, with or without tyrosine, taurine, aspartic acid, and glutamic acid. The average daily total cost of 3CB PN was 74.78–585.82 yuan. Since Some 3CB products had been listed in NMIC, they were affordable for urban patients covered by Urban Employee Basic Medical Insurance but might be economically unacceptable for rural residents who are insured by the Basic Medical Insurance for Urban and Rural Residents and uninsured patients. Conclusions:A variety of 3CB products have been commercially available in China and can meet the individual PN needs of most patients. Although some products have been listed by NMIC, the economic burden can be heavy for certain patient populations.

Objective To explore the diagnostic value of CT radiomics combined with prothrombin induced by vitamin K absence or antagonist-Ⅱ(PIVKA-Ⅱ)for hepatocellular carcinoma in the background of liver cirrhosis.Methods The clinical and CT imaging data of patients with liver cirrhosis were analyzed retrospectively.Hepatocellular carcinoma was observed in the observation group,and no hepatocellular carcinoma was found in the control group.Logistic regression was used to analyze the clinical factors of hepatocellular carcinoma and a Clinic model was constructed.Support vector machine(SVM)was used to construct the optimal feature model(Rad model).An artificial neural network model(Combine model)was built based on Softmax policy using Python3.6.Results The degree of liver cirrhosis,aspartate aminotransferase(AST),alpha-fetoprotein(AFP)and PIVKA-Ⅱ were independent factors for predicting hepatocellular carcinoma(P＜0.05).The sensitivity and specificity of the clinical logistic regression model were 73.15％and 68.34％,respectively.The SVM model was used to construct Radiomics score(Radscore)containing 7 optimal features,and there were significant differences between the two groups(P＜0.001).DeLong test showed that the area under the curve(AUC)of Combine model was significantly higher than that of Rad model and Clinic model(P＜0.05).The Hosmer-Lemeshow test showed that the Combine model agrees well.Decision curve analysis showed that the curves of Combine model were significantly higher than Clinic model,Rad model and the extreme curve.Conclusion The Combine model based on CT radiomics combined with clinical factors can accurately predict the occurrence of hepatocellular carcinoma in the background of liver cirrhosis.

The incidence and mortality rates of hepatocellular carcinoma(HCC)remain high in China,and the application of surgical resection is often limited due to the fact that most patients are in the advanced stage at the time of confirmed diagnosis.This article reviews commonly used advanced locoregional therapies for HCC and the advances in mainstream techniques such as local ablation(radiofrequency ablation,microwave ablation,irreversible electroporation,and cryoablation),intravascular intervention(transcatheter arterial chemoembolization,hepatic arterial infusion chemotherapy,and Y90 hepatic arterial infusion chemotherapy),and radiotherapy(CyberKnife,proton therapy,and heavy-ion therapy),and a multidimensional decision-making framework is constructed for HCC locoregional therapy by comparing treatment principles,indications,limitations,and clinical data of these techniques.This article aims to provide evidence-based support for persistent dilemmas in clinical decision-making,promote the role of locoregional therapies in clinical practice,and propose the directions for future research and clinical application.This article also establishes a comprehensive clinical roadmap for HCC locoregional therapy,which helps to address current challenges regarding technique selection and delineate future directions for innovation,in order to reshape the treatment of HCC through technological integration and paradigm innovation.

Objective To investigate the predictive value of edema metabolic and hematoma dynamics changes on motor and cog-nitive recovery outcomes in patients with intracerebral hemorrhage(ICH).Methods The CT data of ICH patients were collected to evaluate hematoma volume changes from admission to day 3.On day 3,multivoxel magnetic resonance spectroscopy(MRS)was per-formed with region of interest located in the edema region and contralateral normal tissue.Motor and cognitive function recovery was assessed using the simplified F-M scale and the Montreal cognitive assessment(MoCA)on day 3 and at the 3-month follow-up,respec-tively.Overall clinical outcomes were assessed using the Glasgow outcome scale(GOS),and all patients were divided into good and poor outcome groups.Clinical data and metabolic differences in the edema region between the two groups were compared,respec-tively.Logistic regression analysis and receiver operating characteristic(ROC)curves were used to identify and evaluate independent prognostic factors.Subgroup analysis were performed via stratification of hematoma location.Results The logistic regression analy-sis indicated that intraventricular extension,hematoma changes,and the ratio of N-acetyl aspartate(NAA)around the hematoma to contralateral normal brain parenchyma NAA(rNAA)were inde-pendent prognostic factors for poor outcomes(P＜0.05).The area under the curve(AUC)for each factor and the combined model were 0.69,0.73,0.79,and 0.82,respectively.In patients with ICH in the basal ganglia region,△F-M was negatively correlated with hematoma changes and positively correlated with rNAA value(P＜0.001).In patients with ICH in the thalamic and lobar regions,△MoCA was not significantly correlated with hematoma changes(P＞0.05),but was positively correlated with rNAA value(P＜0.001).Conclusion The rNAA holds predictive value for motor and cognitive recovery outcomes following standard treatment.