ObjectiveThis paper aims to investigate the material basis of the anti-inflammatory efficacy and mechanism of action of Bushen Tongdu prescription （BSTDP）. MethodsThe chemical components of BSTDP and its blood-absorbed components in vivo were systematically identified by using ultra-performance liquid chromatography-linear ion trap-electrostatic field orbitrap high-resolution mass spectrometry （UPLC-LIT-Orbitrap-MS）. Network pharmacology was employed to screen blood-absorbed bioactive components and potential targets of this formula. A protein-protein interaction （PPI） network of core targets was constructed to conduct enrichment analysis. Molecular docking was further utilized to verify the binding affinity between key components and targets. The inflammatory model was established and verified in vivo by using a transgenic zebrafish Tg （mpx： GFP）. At three days post-fertilization （3 dpf）， larvae of zebrafish were randomly assigned to blank group， model group， positive drug dexamethasone acetate group （75 μmol·L^-1）， and BSTDP groups with low， medium， and high doses （500， 1 000， and 2 000 mg·L^-1）. The distribution and quantity of neutrophils in the yolk sac region were observed under a fluorescence microscope. The mRNA expression levels of key genes in the toll-like receptor 4 （TLR4）/myeloid differentiation factor 88 （MyD88）/nuclear factor kappa-B （NF-κB） signaling pathway and inflammatory factors including interleukin （IL）-1β， IL-6， and tumor necrosis factor-α （TNF-α） were detected by Real-time quantitative polymerase chain reaction （Real-time PCR）. ResultsA total of 120 chemical components were identified in BSTDP， among which 26 original components were confirmed by using serum pharmacochemical methods. A total of 227 common targets linking rheumatoid arthritis （RA） and the blood-absorbed components were screened by network pharmacology. It is suggested that pseudobrucine， vomicine， sinapine， rehmannioside， cinnamyl alcohol glycoside， and methylephedrine exert anti-inflammatory effects by acting on core targets including protein kinase B1 （Akt1）， signal transducer and activator of transcription 3 （STAT3）， tumor necrosis factor （TNF）， TLR4， mitogen-activated protein kinase 14 （MAPK14）， and phosphatidylinositol-4，5-bisphosphate 3-kinase catalytic subunit α （PIK3CA）， thereby modulating multiple signaling pathways such as TLR4 and NF-κB. In vivo verification in zebrafish demonstrates that the maximum tolerable concentration of Bushen Tongdu Formula is 2 000 mg·L^-1. Compared to those in the blank group， zebrafish in the model group showed a significantly higher number of neutrophils in the yolk sac region （P<0.01） and rising mRNA levels of TLR4， MyD88， NF-κB， TNF-α， IL-6， and IL-1β （P<0.01）. Compared to that in the model group， the number of neutrophils was significantly reduced in BSTDP groups with medium and high doses， as well as the dexamethasone acetate group （P<0.05， P<0.01）. There was no statistically significant difference in the low dose group. The mRNA expression levels of TLR4， MyD88， NF-κB， TNF-α， IL-6， and IL-1β were significantly down-regulated （P<0.05， P<0.01）. ConclusionThis paper identifies the material basis of the efficacy of BSTDP， demonstrating that the formula can exert an anti-inflammatory effect through the TLR4/MyD88/NF-κB signaling pathway. The results provide scientific experimental evidence for its further clinical application.

ObjectiveThis paper aims to investigate the material basis of the anti-inflammatory efficacy and mechanism of action of Bushen Tongdu prescription （BSTDP）. MethodsThe chemical components of BSTDP and its blood-absorbed components in vivo were systematically identified by using ultra-performance liquid chromatography-linear ion trap-electrostatic field orbitrap high-resolution mass spectrometry （UPLC-LIT-Orbitrap-MS）. Network pharmacology was employed to screen blood-absorbed bioactive components and potential targets of this formula. A protein-protein interaction （PPI） network of core targets was constructed to conduct enrichment analysis. Molecular docking was further utilized to verify the binding affinity between key components and targets. The inflammatory model was established and verified in vivo by using a transgenic zebrafish Tg （mpx： GFP）. At three days post-fertilization （3 dpf）， larvae of zebrafish were randomly assigned to blank group， model group， positive drug dexamethasone acetate group （75 μmol·L^-1）， and BSTDP groups with low， medium， and high doses （500， 1 000， and 2 000 mg·L^-1）. The distribution and quantity of neutrophils in the yolk sac region were observed under a fluorescence microscope. The mRNA expression levels of key genes in the toll-like receptor 4 （TLR4）/myeloid differentiation factor 88 （MyD88）/nuclear factor kappa-B （NF-κB） signaling pathway and inflammatory factors including interleukin （IL）-1β， IL-6， and tumor necrosis factor-α （TNF-α） were detected by Real-time quantitative polymerase chain reaction （Real-time PCR）. ResultsA total of 120 chemical components were identified in BSTDP， among which 26 original components were confirmed by using serum pharmacochemical methods. A total of 227 common targets linking rheumatoid arthritis （RA） and the blood-absorbed components were screened by network pharmacology. It is suggested that pseudobrucine， vomicine， sinapine， rehmannioside， cinnamyl alcohol glycoside， and methylephedrine exert anti-inflammatory effects by acting on core targets including protein kinase B1 （Akt1）， signal transducer and activator of transcription 3 （STAT3）， tumor necrosis factor （TNF）， TLR4， mitogen-activated protein kinase 14 （MAPK14）， and phosphatidylinositol-4，5-bisphosphate 3-kinase catalytic subunit α （PIK3CA）， thereby modulating multiple signaling pathways such as TLR4 and NF-κB. In vivo verification in zebrafish demonstrates that the maximum tolerable concentration of Bushen Tongdu Formula is 2 000 mg·L^-1. Compared to those in the blank group， zebrafish in the model group showed a significantly higher number of neutrophils in the yolk sac region （P<0.01） and rising mRNA levels of TLR4， MyD88， NF-κB， TNF-α， IL-6， and IL-1β （P<0.01）. Compared to that in the model group， the number of neutrophils was significantly reduced in BSTDP groups with medium and high doses， as well as the dexamethasone acetate group （P<0.05， P<0.01）. There was no statistically significant difference in the low dose group. The mRNA expression levels of TLR4， MyD88， NF-κB， TNF-α， IL-6， and IL-1β were significantly down-regulated （P<0.05， P<0.01）. ConclusionThis paper identifies the material basis of the efficacy of BSTDP， demonstrating that the formula can exert an anti-inflammatory effect through the TLR4/MyD88/NF-κB signaling pathway. The results provide scientific experimental evidence for its further clinical application.

ObjectiveTo explore the clinical significance of artemin （ARTN） expression in prostate cancer （PCa） tissues and its impact on the malignant behavior of PCa cell lines. MethodsImmunohistochemistry was used to detect the expression of ARTN protein in 40 benign prostate tissues and 91 PCa tissues， and its relationship with the clinical and pathological characteristics of PCa was analyzed. PCa stable cell lines with ARTN knockdown were constructed， and the effects of ARTN on the proliferation， migration， and invasion ability of PCa cells were detected via CCK-8 cell proliferation assay and Transwell assay. Western blot assay was used to detect the effect of ARTN on the expression of epithelial-mesenchymal transition （EMT） related markers E-cadherin， N-cadherin， Vimentin， and Snail family transcription inhibitory factor 1 （Snail-1）. ResultsARTN was highly expressed in PCa and correlated with Gleason score， local lymph node metastasis， and local nerve invasion （P0.05）. Survival analysis showed a statistically significant difference in survival rates between ARTN positive and negative patients （P=0.027）. The results of CCK-8 and Transwell assay showed that the knockdown of ARTN could inhibit the proliferation， migration， and invasion ability of PCa cells （all P0.05）. Western blot results showed that the knockdown of ARTN upregulated the epithelial marker E-cadherin in PCa cells， while the mesenchymal markers N-cadherin， Snail-1， and Vimentin were downregulated. ConclusionARTN is highly expressed in PCa and can promote the proliferation， migration， invasion ability of PCa cells， as well as increasing EMT levels in PCa cells， suggesting it may be a potential target for the diagnosis and treatment of PCa.

Objective To specifically extract and analyze nascent proteins synthesized by bone marrow mesenchymal stem cells(BMSCs)after transplantation into ischemic hearts using a technique employing mutant methionyl-tRNA synthetase(MetRSL247G)for nascent protein labeling,in order to explore the potential mechanisms of action in BMSCs post-transplantation.Methods Point mutation at position 274 of the MetRS gene in BMSCs was induced via lentiviral infection to enable azidonorleucine(ANL)-mediated labeling of nascent proteins in BMSCs.The labeling efficiency was verified by means of fluorescent non-canonical amino-acid tagging(FUNCAT).Thirty healthy female C57BL/6J mice(8-10 weeks old)were divided into control and experimental groups,with 15 mice in each group.The acute myocardial infarction model was constructed by ligating the left anterior descending coronary artery in experimental group,while control mice underwent only thoracotomy without coronary ligation.After modeling,both groups received intramyocardial injections of MetRSL247G-modified BMSCs(MetRSL247G-BMSCs)at 3 different sites in the peri-infarct ischemic region.Mice were intraperitoneally injected with ANL every 6 hours for 4 times on postoperative days 0,2,and 6(n=5 for each time point)respectively,euthanized 24 h after the last injection,and cardiac tissues were isolated.The newly synthesized and labeled proteins produced by BMSCs after transplantation into the myocardium of experimental and control groups were collected,using an enrichment technique for ANL-tagged proteins and liquid chromatography-tandem mass spectrometry(LC-MS)analysis.Gene ontology(GO)analysis,Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis,protein-protein interaction(PPI)analysis,and heatmap visualization analysis were performed to identify differentially expressed proteins at the 3 time points and screen key pathways and genes.Results Under fluorescence microscopy,the MetRSL247G lentivirus-infected BMSCs were observed to be labelled with mCherry signals,confirming the successful construction of the MetRSL247G-BMSCs cell line.Green fluorescent signals were detected only in nascent proteins in culture medium containing both MetRSL247G-BMSCs and ANL,validating the sensitivity and specificity of the labeling method.GO analysis revealed that differentially expressed proteins were primarily involved in basic cellular biological processes such as extracellular exosome formation,extracellular matrix organization,and focal adhesion.KEGG and PPI analyses indicated that the differential proteins were mainly involved in complement and coagulation cascade pathway,actin cytoskeleton regulation pathway,and apoptosis pathway.Heatmap analysis showed significantly upregulated expression of anti-apoptosis and cell adhesion-related factors in experimental group on day 1(P＜0.05),upregulated anti-apoptotic factors,pro-apoptotic factors,and cell adhesion-related factors on day 3(P＜0.05),and upregulated anti-apoptotic factors,cell differentiation-related factors,and cell adhesion-related factors on day 7(P＜0.05)compared with control group.Expression of apoptosis-inducing factor 1 was significantly downregulated on days 1 and 7(P＜0.05).On day 3,most differentially expressed proteins,including anti-apoptosis factors(Protein S100-A11,Clusterin,Gelsolin),pro-apoptosis factor(Cathepsin B),cell differentiation-related factor(Transgelin-2),and cell adhesion-related factors(Cofilin-1,Periostin,Fibronectin)were significantly upregulated(P＜0.05).Conclusions The MetRSL247G mutation enables BMSCs to incorporate ANL and synthesize labeled proteins,confirming the feasibility of this nascent protein labeling technique.Nascent proteins of BMSCs in ischemic myocardium primarily contribute to extracellular exosome secretion and extracellular matrix organization.BMSCs may adapt to and respond to ischemic and hypoxic environments by influencing complement and coagulation cascades,activating inflammatory factors,regulating actin cytoskeleton structure,and modulating apoptosis,thereby maintaining the survival of BMSCs.

INTRODUCTION: Lecanemab has shown promise in treating early Alzheimer's disease (AD), but its safety and efficacy in Chinese populations remain unexplored. This study aimed to evaluate the safety and 6-month clinical outcomes of lecanemab in Chinese patients with mild cognitive impairment (MCI) or mild AD. METHODS: In this single-arm, real-world study, participants with MCI due to AD or mild AD received biweekly intravenous lecanemab (10 mg/kg). The study was conducted at Hainan Branch, Ruijin Hospital Shanghai Jiao Tong University School of Medicine. Patient enrollment and baseline assessments commenced in November 2023. Safety assessments included monitoring for amyloid-related imaging abnormalities (ARIA) and other adverse events. Clinical and biomarker changes from baseline to 6 months were evaluated using cognitive scales (mini-mental state examination [MMSE], montreal cognitive assessment [MoCA], clinical dementia rating-sum of boxes [CDR-SB]), plasma biomarker analysis, and advanced neuroimaging. RESULTS: A total of 64 patients were enrolled in this ongoing real-world study. Safety analysis revealed predominantly mild adverse events, with infusion-related reactions (20.3%, 13/64) being the most common. Of these, 69.2% (9/13) occurred during the initial infusion and 84.6% (11/13) did not recur. ARIA-H (microhemorrhages/superficial siderosis) and ARIA-E (edema/effusion) were observed in 9.4% (6/64) and 3.1% (2/64) of participants, respectively, with only two symptomatic cases (one ARIA-E presenting with headache and one ARIA-H with visual disturbances). After 6 months of treatment, cognitive scores remained stable compared to baseline (MMSE: 22.33 ± 5.58 vs . 21.27 ± 4.30, P = 0.733; MoCA: 16.38 ± 6.67 vs . 15.90 ± 4.78, P = 0.785; CDR-SB: 2.30 ± 1.65 vs . 3.16 ± 1.72, P = 0.357), while significantly increasing plasma amyloid-β 42 (Aβ42) (+21.42%) and Aβ40 (+23.53%) levels compared to baseline. CONCLUSIONS: Lecanemab demonstrated a favorable safety profile in Chinese patients with early AD. Cognitive stability and biomarker changes over 6 months suggest potential efficacy, though high dropout rates and absence of a control group warrant cautious interpretation. These findings provide preliminary real-world evidence for lecanemab's use in China, supporting further investigation in larger controlled studies. REGISTRATION ClinicalTrials.gov , NCT07034222.
Humans ; Alzheimer Disease/drug therapy* ; Male ; Female ; Aged ; Middle Aged ; Cognitive Dysfunction/drug therapy* ; Aged, 80 and over ; Amyloid beta-Peptides/metabolism* ; Biomarkers ; East Asian People

ObjectiveTo investigate the effects of foliar fertilizer of nano-calcium carbonate and calcium nitrate tetrahydrate on the agronomic traits， carbohydrate and endogenous hormone contents of Dendrobium officinale planted for 1 year under greenhouse cultivation， in order to provide scientific basis for fertilization to improve the yield and quality of D. officinale. MethodsSingle-factor experimental design was adopted. Starting from early spring， D. officinale was treated with foliar spraying according to corresponding fertilizers. Three treatment groups were established based on different fertilizers， namely， a blank group（clear water）， a nano-calcium carbonate group（0.727 g·L^-1 nano-calcium carbonate water-soluble fertilizer）， and a calcium nitrate tetrahydrate group（1.091 g·L^-1 calcium nitrate tetrahydrate water-soluble fertilizer）. The frequency of spraying was three times per month， and the entire treatment process lasted for nine months. The effects of various treatments on the traits and relative chlorophyll content of D. officinale were dynamically monitored. Sampling was conducted at three specific time points：August 2， 2023， September 8， 2023， and November 1， 2023， respectively. The contents of glucose and mannose in D. officinale stems were determined by high performance liquid chromatography（HPLC）， the content of soluble sugars in D. officinale stems and leaves was determined by phenol method， and enzyme-linked immunosorbent assay（ELISA） was used to detect the concentrations of cytokinin and auxin. ResultsCompared with the blank group， the treatments with nano-calcium carbonate and calcium nitrate tetrahydrate could significantly increase stem length， stem node number， leaf number， and tiller number. Among them， during the harvesting period in November， the stem length and tiller number， which are indicators related to the yield of D. officinale， increased by 60.85% and 19.23% after treatment with calcium nitrate tetrahydrate， and by 32.54% and 28.85% after treatment with nano-calcium carbonate， respectively. Compared with the blank group， treatments with nano-calcium carbonate and calcium nitrate tetrahydrate could promote the accumulation of sucrose in the stems and leaves of D. officinale to varying degrees， as well as the accumulation of polysaccharides， mannose， and glucose in the stems. In addition， nano-calcium carbonate treatment also facilitated the accumulation of fructose in the stems and leaves of D. officinale. Specifically， during the harvesting period in November， polysaccharides and mannose， which were the main active ingredients in D. officinale stems， increased by 28.48% and 29.36% after treatment with calcium nitrate tetrahydrate， and by 39.91% and 82.62% after treatment with nano-calcium carbonate， respectively. In addition， compared with the blank group， the concentrations of auxin in the stems and leaves of D. officinale were significantly increased after treatment with calcium nitrate tetrahydrate（P<0.05）. Similarly， the concentrations of cytokinin and auxin in the stems of D. officinale were also elevated after treatment with nano-calcium carbonate. Correlation analysis further indicated that elongation growth and tillering of D. officinale stems after foliar spraying of nano-calcium carbonate and calcium nitrate tetrahydrate might be related to the accumulation of carbohydrates in the stems and leaves and the synergistic effect of auxin and cytokinin. ConclusionIn production practice， spraying nano-calcium carbonate and calcium nitrate tetrahydrate can promote the accumulation of cytokinin， auxin， and carbohydrate contents in the stems and leaves of D. officinale， and promote tillering and elongation growth of the stems.

Aim To investigate the mechanisms of the ultrafiltration membrane extract of Angelica sinensis and Radix Hedysari extracts on oxidative stress in rats with diabetic kidney disease(DKD).Methods Forty-five SD rats were randomly divided into a control group(n=8)a model group(n=37).Rats in the model group were fed a high-sugar,high-fat diet for four weeks,followed by intraperitoneal injection of strepto-zotocin at a dose of 30 mg·kg-1 to induce diabetes in the rats.Three weeks later,rats with 24-hour urinary protein(24-hUP)levels more than or equal to 30 mg were injected via the tail vein with 0.05 mg·kg-1 of 10％high molecular weight dextran for three times to induce a model of blood stasis in diabetic kidney dis-ease(DKD).The rats were then evaluated for random blood glucose(GLU)levels,24-hUP,biochemical markers,histopathological staining,and the protein expression of nicotinamide adenine dinucleotide phos-phate(NADPH)oxidase(NOX)1,NOX2,NOX3,NOX4,and NOX5 in renal tissues using immunoblot a-nalysis.Results Compared to the control group,rats in the model group showed significantly increased GLU,24-hUP,SCr,BUN,TG,TC,bu markedly de-creased ALB2,HDL,LDL levels,and the relative ex-pression of NOX1,NOX2,NOX4,NOX5 proteins in-creased markedly(P＜0.01);Comparison with the model group,rats in the treatment group exhibited sig-nificantly decreased GLU,24-hUP,SCr,BUN,TG,TC at 6 weeks and 8 weeks,but markedly increased ALB2,HDL,LDL levels,and the relative expression of NOX1,NOX2,NOX4,NOX5 proteins decreased significantly(P＜0.05).Conclusions The ultrafil-tration membrane extract of Angelica sinensis and Radix Hedysari can effectively ameliorate oxidative stress and renal function in DKD rats,which may be associated with targets within the NOX family.

Objective To clarify the effect of preoperative oral carbohydrate on overweight or obese patients undergoing laparoscopic hysterectomy in Trendelenburg position.Methods A total of 72 overweight or obese patients who underwent laparoscopic hysterectomy in Trendelenburg position at Yancheng Third People's Hospital from September 2022 to August 2024 were selected,and randomly divided into the control group and the observation group,with 36 cases in each group.The control group received routine restriction of fasting and fluid starting from 22:00 on the day before surgery,while the observation group was given an additional 300 mL of carbohydrate orally 2 hours before anesthesia on the basis of the control group.The preoperative ultrasound results of gastric antrum,Perlas classification,full stomach status,visual analogue scale(VAS)scores for thirst and hunger,awakening time,tracheal extubation time,intraoperative hypothermia,gastro-intestinal function recovery time,and adverse reactions were compared between the two groups.Results Before anesthesia,there was no statistically significant difference in the cross-sectional area of the gastric antrum(CSA)in the right lateral position,gastric volume(GV),or the ratio of gastric volume to body weight(GV/W)between the two groups(P>0.05).There was also no statistically significant difference in the Perlas classification of gastric antrum ultrasound or the incidence of full stomach between the two groups(P>0.05).Immediately before anesthesia,the VAS scores for thirst and hunger in the observation group were significantly lower than those in the control group(P<0.05).The awakening time,tracheal extubation time,and incidence of intraoperative hypothermia in the observation group were significantly shorter/lower than those in the control group(P<0.05).The observation group had significantly shorter time of first exhaust and defecation after surgery,as well as lower incidence of postoperative bloating,compared with the control group(P<0.05).Conclusion For overweight or obese patients undergoing laparoscopic hysterectomy in the Trendelenburg position,oral intake of 300 mL carbohydrate 2 hours before anesthesia does not increase the risk of regurgitation or aspiration,with safety and reliability.It not only can improve comfort of patients and promote post-anesthesia awakening,but also can reduce the incidences of intraoperative hypothermia and gastrointestinal adverse reactions,which is conducive to accelerating postoperative recovery.

Objective To analyze differences of purified water,softened water,and concentrated softened water on the decontamination effects for surgical instruments,so as to provide a reliable reference for the selection of cleaning water for surgical instruments.Methods The physical and chemical indexes and components of concentrated softened water,softened water,and purified water were detected,and their cleaning effects for instrument were compared.The decontamination cleaning experiment was conducted in three hospitals with different types of cleaning water using mini cleaning machines,and the differences in the decontamination time were analyzed.The cleaning and decontamination experiments on contaminated instruments was performed using a spray cleaning and disinfection device,and the cleaning effects of instruments with different types of cleaning water were analyzed.Results Calcium content and hardness of the three types of cleaning water were all at a low level,which can avoid scale adhesion for cleaning surgical instruments.There were statistically significant differences in the pH value,conductivity,and contents of calcium,sodium and chloride among the three cleaning waters(P<0.05).The purified water was weakly acidic,and its conductivity,hardness,and contents of calcium,sodium and chloride were all at low levels.The softened water and concentrated softened water were weakly alkaline,with high levels of conductivity and sodium content and low level of chloride.There were statistically significant differences in the decontamination time of the purified water,softened water and concentrated softened water among hospitals(P<0.05).Under the same contamination condition of surgical instruments,there was a statistically significant difference in the qualified rate of instruments cleaning in the upper and lower cleaning baskets with different cleaning waters(P<0.05).Conclusion Concentrated softened water and softened water have high sodium content,and their decontamination and cleaning abilities are significantly stronger than those of the purified water.Purified water has poorer effects in instruments cleaning for its deionized property.Using concentrated softened water or softened water in the surgical instrument cleaning process can achieve effects of reducing consumption and increasing efficiency,which is conducive to improving cleaning efficiency and quality.

Aim To establish an animal model of diabetic kidney disease(DKD)integrating blood stasis syndrome and syndrome evaluation indicators.Methods Twenty-five SD rats were ran-domly divided according to body weight into a control group(8 rats)and a modeling group(17 rats).The modeling group was fed a high-sugar and high-fat diet for four weeks and induced to form diabetic rats by intraperitoneal injection of 30 mg·kg-1 streptozocin.The modeling rats were randomly divided into the DKD group and blood stasis syndrome combination group accord-ing to 24-hour urinary protein(24-hUP).The blood stasis syn-drome combination group was induced to replicate the DKD blood stasis syndrome model by injecting 10％high molecular weight D-glucoside three times at a dose of 0.05 mg·kg-1 via tail vein.The model was evaluated based on random blood glu-cose level,24-hUP level,syndrome assessment,pathological staining etc,and differential metabolites were selected using metabolomics.Results The comprehensive evaluation of syn-drome manifestations and pathological staining in the combined model of blood stasis syndrome in rats demonstrated successful replication.Utilizing the technique of liquid chromatography-mass spectrometry,22 differential metabolites were identified,with associated pathways showing a certain relevance to blood stasis syndrome in DKD.Conclusions The successful replica-tion of an animal model combining the syndrome of blood stasis in DKD has been achieved in this study.Evaluation of indicators and results from metabolomics studies consistently demonstrate a correlation with the syndrome of blood stasis in DKD.