ObjectiveTo investigate whether Huanglian Jiedutang （HLJD） and its major active constituents （geniposide， baicalin， and berberine） can inhibit the inflammatory response of BV2 cells under lipopolysaccharide （LPS） stimulation via the high-mobility group protein B1 （HMGB1）/Toll-like receptor 4 （TLR4）/nuclear factor-κB （NF-κB） signaling pathway， and to explore differences in therapeutic efficacy among the three monomers， their combined formula， and HLJD under equal content ratios. MethodsBV2 microglial cells were used as the primary experimental model. Cell viability was assessed using the cell counting kit-8 （CCK-8） method to examine the effects of different concentrations of dimethyl sulfoxide （DMSO， 0.8%， 0.4%， 0.2%， 0.1%， and 0.05%） on cell viability. IncuCyte was employed to monitor the growth of cells under different concentrations of HLJD （200， 100， 50， 25， 12.5， 6.25 mg·L^-1）. Nitric oxide （NO） assay was used to screen the optimal HLJD concentration. High-performance liquid chromatography （HPLC） determined the content of geniposide， baicalin， and berberine in HLJD， and experimental groups were subsequently established according to the relative proportions of these constituents. CCK-8 assay evaluated cell viability under different treatments. Enzyme-linked immunosorbent assay （ELISA） measured levels of inflammatory factors （TNF-α， IL-1β， IL-6， IL-10） in the supernatant. Flow cytometry assessed the effects of treatments on M1-type polarization of BV2 cells. Western blot determined the expression levels of HMGB1， TLR4， and NF-κB-related proteins. ResultsCompared with the blank group， DMSO at concentrations ≤0.2% did not affect cell viability within 48 h. BV2 cell growth plateaued at 24 h after treatment with 200 mg·L^-1 HLJD. Under stimulation with 2 mg·L^-1 LPS， this concentration of HLJD effectively reduced NO release， and 6 h pre-treatment had a stronger inhibitory effect on NO than direct administration. HPLC results showed that 1 mg of HLJD freeze-dried powder contained approximately 24 μg of geniposide， 15 μg of baicalin， and 30 μg of berberine. Based on these ratios， experimental groups were blank， LPS （2 mg·L^-1）， HLJD （200 mg·L^-1）， monomer combination， geniposide （4.8 mg·L^-1）， baicalin （3 mg·L^-1）， and berberine （6 mg·L^-1）. The monomer combination group consisted of all three active constituents dissolved together. LPS and HLJD or its active constituents did not affect cell viability compared with the blank group. LPS significantly increased TNF-α， IL-1β， IL-6， and IL-10 in the supernatant （P<0.01）. HLJD and its active constituents significantly reduced pro-inflammatory factors TNF-α， IL-1β， and IL-6 （P<0.05， P<0.01） while upregulating anti-inflammatory IL-10 （P<0.01）， with the monomer combination showing the strongest effect （P<0.05， P<0.01）. Compared with the blank group， LPS significantly increased the proportion of CD80⁺CD86⁺ （M1-type） BV2 cells （P<0.01）. HLJD and its constituents partially inhibited M1 polarization （P<0.05， P<0.01）， with the monomer combination exhibiting the most pronounced effect （P<0.05， P<0.01）. Compared with the blank group， LPS upregulated HMGB1， TLR4， and NF-κB-related proteins （P<0.01）， whereas HLJD and its active constituents significantly reduced their expression （P<0.05， P<0.01）， with the monomer combination having the strongest regulatory effect （P<0.05， P<0.01）. ConclusionHLJD and its major active constituents （geniposide， baicalin， berberine） can inhibit LPS-induced inflammatory responses in BV2 cells. The combination of the three active constituents demonstrates the most potent anti-inflammatory effect， significantly attenuating M1-type polarization of BV2 cells via the HMGB1/TLR4/NF-κB signaling pathway.

Diabetes mellitus, characterized by glucose metabolism disorders and marked by insulin deficiency or insulin resistance, has seen a continuous rise in prevalence. In recent years, islet transplantation has matured as a therapeutic approach for diabetes, becoming an important method for glycemic control and the reduction of diabetes-related complications. Donor selection directly influences transplant outcomes, and various research institutions worldwide have proposed multiple scoring systems to optimize donor assessment, such as the University of Alberta scoring system and the North American Islet Donor Score. This article explores the impact of key factors such as donor age, body mass index and ischemia time on islet transplantation. Combining practical experience in pancreatic donor selection from Shanghai Changzheng Hospital, it proposes screening criteria for pancreatic donors suitable for China, aiming to provide new evidence for improving the success rate of islet transplantation.

Bacterial infection is a major threat to global public health,and can cause serious diseases such as bacterial skin infection and foodborne diseases.It is essential to develop a new method to rapidly di-agnose clinical multiple bacterial infections and monitor food microbial contamination in production sites in real-time.In this work,we developed a 4-mercaptophenylboronic acid gold nanoparticles(4-MPBA-AuNPs)-functionalized hydrogel microneedle(MPBA-H-MN)for bacteria detection in skin inter-stitial fluid.MPBA-H-MN could conveniently capture and enrich a variety of bacteria within 5 min.Surface enhanced Raman spectroscopy(SERS)detection was then performed and combined with ma-chine learning technology to distinguish and identify a variety of bacteria.Overall,the capture efficiency of this method exceeded 50％.In the concentration range of 1 × 10 7 to 1 × 10 10 colony-forming units/mL(CFU/mL),the corresponding SERS intensity showed a certain linear relationship with the bacterial concentration.Using random forest(RF)-based machine learning,bacteria were effectively distinguished with an accuracy of 97.87％.In addition,the harmless disposal of used MNs by photothermal ablation was convenient,environmentally friendly,and inexpensive.This technique provided a potential method for rapid and real-time diagnosis of multiple clinical bacterial infections and for monitoring microbial contamination of food in production sites.

Currently,there are practical and technical difficulties in the construction of clinical ques-tions in the development of clinical practice guidelines.Clinicians or guideline developers seldom construct clin-ical questions based the actual case scenario,leading to some information loss between structured and actual clinical connotation.To overcome this challenge,we proposed a case-guided questions construction approach,and carried out case research and verification in the formulation of the guideline.We found that this method could more efficiently and scientifically assist the formulation of clinical questions,and provide reference for clinicians or guideline developers.

Objective To explore the potential causal relationship between opioids and chronic renal failure(CRF)using Mendelian randomization analysis.Methods Screening for single nucleotide polymorphisms(SNPs)strongly associated with opioids and CRF firstly,causality was analyzed by using two-sample Mendelian randomization methods,heterogeneity,horizontal pleiotropy and sensitivity analyses were then performed.Results A total of 41 opioid-related SNPs were screened in this study,inverse variance weighting analysis suggested that opioid use is a risk factor for CRF(OR=1.15,95％CI:1.04-1.27,P＜0.01).The heterogeneity test showed that there was no heterogeneity in the SNPs,and the horizontal multivariate analysis and sensitivity analysis confirmed the robustness of the results.Conclusion Genetically,the study suggests a positive causal association between opioids and CRF,opioid use is a risk factor for CRF.

Objective Under the guidance of nurses,an intelligent medical waste temporary storage equipment was developed and its application effect was evaluated,aiming to optimize the medical waste recycling process and save human resources.Methods A research and development(R&D)team was established to analyze the issues in the current medical waste temporary storage process.An intelligent medical waste temporary storage device was developed,equipped with functions such as categorized disposal,permission scanning,barcode recognition,intelligent weighing,overflow alarm,and ozone disinfection.From January 19th to February 20th,2023,an intelligent medical waste storage equipment was placed in the medical waste storage room on the 7th to 9th floors of the outpatient clinic of a tertiary hospital in Wuhan city as an experimental group;traditional medical waste storage bins were placed on the 4th and 5th floors of the outpatient department as a control group.The daily disposal indicators and environmental hygiene monitoring indicators of 2 groups of medical waste were compared.Results With the application of the intelligent medical waste temporary storage device,the disinfection rate of medical waste increased from 5.00％to 100％;the overflow rate decreased from 42.52％to 0％;the handover time reduced from 4.98±2.21 minutes to 1.07±0.35 minutes;the standardization rate of handovers rose from 15.83％to 100％.All these differences were statistically significant(P＜0.001).Additionally,the pass rate of bacterial colonies on the inner walls of bins increased from 53.33％to 67.78％,with a statistically significant difference(P=0.012).Conclusion The application of the intelligent medical waste temporary storage equipment significantly improves the efficiency and quality of medical waste management;at the same time,it also plays an active role in improving the storage environment,which can safeguard the environmental hygiene of hospitals and the safety of patients.

Objective To investigate the mechanism of Wenhe Decoction in the treatment of febrile seizures through network pharmacology based on NLRP3/Caspase-1/GSDMD signaling pathway;To conduct experimental verification.Methods The active components and targets of Wenhe Decoction were retrieved and screened through TCMSP,BATMAN-TCM,PubChem databases and SwissADME platform.The disease targets of febrile seizures were found in GenCards,OMIM and DisGeNET databases.The intersection targets of Wenhe Decoction and the disease and the active components corresponding to the intersection targets were imported into Cytoscape 3.7.2 software to construct the Chinese materia medica-active components-targets network.The intersection targets were submitted to the STRING database to construct a protein-protein interaction network,and then the intersection targets were imported into the Metascape database for GO and KEGG pathway enrichment analysis.The febrile seizures rat model was established,and Wenhe Decoction of 4.05,8.1 and 16.2 g/kg were given respectively by gavage for 21 days.The rats were placed in batches in(45±0.5)℃constant temperature water bath to induce convulsive seizures,and the convulsive latency time and convulsive duration of rats were recorded.The behavioral differences of mice were observed.The morphology of hippocampal tissue were observed by HE and Nissl staining.The ROS content of hippocampal tissue was detected by DHE fluorescent probe technology.The serum ATP,GABA,Glu,Caspase-1,GSDMD,IL-1β and IL-18 contents were detected by ELISA,and the expression of NLRP3 inflammasome-related protein in hippocampal tissue was detected by Western blot.Results Network pharmacology analysis obtained 98 active components of Wenhe Decoction and 1 838 targets.162 targets were obtained by intersecting with disease targets,the core components for the treatment of febrile seizures were β-sitosterol,quercetin,luteolin,trans-squalene,sitosterol,saponin,etc.,and the core targets were EGFR,TNF,JUN,MTOR,etc.,and mainly through the regulation of inflammatory response,apoptosis,mitochondrial function and energy metabolism,mediating anti-inflammatory pathways such as PI3K-Akt signaling pathway and calcium signaling pathway to exert anticonvulsant effects.The experimental results showed that Wenhe Decoction could prolong the convulsive latency and shorten the duration of convulsions in febrile seizures model rats,decrease the level of convulsions,and the pathological damage of hippocampal tissue was improved and damaged neurons were repaired.The serum contents of ROS,Glu,Caspase-1,GSDMD,IL-1β and IL-18 were significantly reduced,and ATP and GABA contents significantly increased.The protein expressions of NLRP3,ASC,pro-Caspase-1,pro-IL-1β and pro-IL-18 in hippocampal tissue significantly decreased.Conclusion Wenhe Decoction may intervene in febrile seizures rats through NLRP3/Caspase-1/GSDMD signaling pathway,inhibit pyroptosis to reduce the occurrence of neuroinflammation,and then affect the balance of neurotransmitters Glu and GABA to play a role in anti-febrile seizures and prevent brain tissue damage.

The incidence of delayed chemotherapy-induced nausea and vomiting is relatively high among pediatric cancer patients. Nausea and vomiting symptoms can exacerbate physical and psychological burdens, potentially leading to aversion and reduced treatment adherence. This paper analyzes and summarizes delayed chemotherapy-induced nausea and vomiting in pediatric cancer patients, covering overview, influencing factors, assessment tools, and non-pharmacological interventions, aiming to provide insights for clinical prevention and intervention strategies targeting delayed chemotherapy-induced nausea and vomiting in pediatric patients.

Objective:To explore the effects of miR-125b on the proliferation, invasion and migration of pancreatic cancer cells by targeting SMYD2 signaling pathway.Methods:The expression of miRNA-125b in Aspc-1 and BxPC-3 lines of pancreatic cancer cells were detected. miRDB, ENCORI and TargetScan databases were used to predict the potential target genes of miRNA-125b. The downstream target genes of miRNA-125b were identified by qPCR assay and double luciferase reporter gene assay. Western blot analysis was performed to detect SMYD2 protein expression after transfection with miRNA-125b inhibitor. EdU staining, Annexin V-FITC/PI assay and Annexin V-FITC/PI assay were used to detect the effects of miRNA-125b inhibitor transfection and simultaneous transfection of miRNA-125b and SMYD2 inhibitor on cell proliferation, clonogenesis and apoptosis.Results:The expression level of miRNA-125b in pancreatic cancer cell lines was higher than that in normal pancreatic duct cells ( P<0.05). The downstream target gene of miRNA-125b was identified as SMYD2 by qPCR assay and double luciferase reporter gene assay. The expression of SMYD2 protein in miR-125b inhibitor group was higher than that in NC group ( P<0.01). EdU cell proliferation assay showed that the number of miRNA-125b positive cells in inhibitor group was lower than that in NC group and Inhibitor NC group ( P<0.05). The number of clones in miR-125b inhibitor+si-SMYD2 group was more than that in miR-125b inhibitor group ( P<0.01). Annexin V-FITC/PI assay showed that the apoptosis number of cell cells in miR-125b inhibitor+si-SMYD2 group was lower than that in miR-125b inhibitor group ( P<0.01) . Conclusion:miRNA-125b is highly expressed in pancreatic cancer cells, and can directly affect the expression of SMYD2 gene, thereby promoting the proliferation and inhibiting the apoptosis of pancreatic cancer cells.

Objective:To evaluate the role of YTH domain family protein 2 (YTHDF2) in myocardial ischaemia-reperfusion injury (MIRI) in diabetic rats and the relationship with the nuclear factor E2-related factor 2 (NRF2)-ferritinophagy.Methods:This experiment was performed in 2 parts. Part Ⅰ Animal experiment SPF healthy male rats, aged 6-8 weeks, weighing 200-220 g, were used. A type 1 diabetes mellitus (DM) model was established by intraperitoneal injection of 1% streptozotocin at a dose of 65 mg/kg. Thirty-six diabetic rats were divided into 3 groups ( n=12 each) using a random number table method: DM sham operation group (DS group), DM myocardial ischaemia-reperfusion group (DIR group), and YTHDF2 knockdown + DM myocardial ischaemia-reperfusion group (AAV-Y+ DIR group). Another 36 non-diabetic rats were selected and divided into 4 groups using the random number table method: sham operation group (NS group, n=12), myocardial ischaemia-reperfusion group (NIR group, n=12), adeno-associated virus control group (AAV-N group, n=6), and YTHDF2 knockdown group (AAV-Y group, n=6). The MIRI model was established by ligating the left anterior descending branch of the coronary artery for 30 min, followed by reperfusion for 2 h. Adeno-associated virus was employed to knock down YTHDF2. At the end of reperfusion, serum concentrations of creatine kinase isoenzyme MB(CK-MB) and cardiac troponin Ⅰ(cTnI) were measured using enzyme-linked immunosorbent assay. The animals were sacrificed, myocardial tissues were harvested, and the pathological changes were observed with a light microscope to assess the myocardial infarct size. The expression of YTHDF2, NRF2, and nuclear receptor coactivator 4 (NCOA4) was detected by Western blot. Part Ⅱ Cell experiment H9c2 cells were divided into 9 groups ( n=24 each) using the random number table method: control group (NC group), high-glucose group (HG group), hypoxia-reoxygenation group (HR group), high-glucose hypoxia-reoxygenation group (HHR group), transfection control group (siN group), YTHDF2 knockdown group (siY group), YTHDF2 knockdown + high-glucose hypoxia-reoxygenation group (siY + HHR group), NRF2 inhibitor ML385 + high-glucose hypoxia-reoxygenation group (M + HHR group), and YTHDF2 knockdown + NRF2 inhibitor ML385 + high-glucose hypoxia-reoxygenation group (siY + M + HHR group). The cells were transfected with siRNA to knock down YTHDF2, and a high-glucose, hypoxia and reoxygenation injury model was established by subjecting cells to 48 h of high glucose, followed by 4 h of hypoxia and 2 h of reoxygenation. The cell viability and lactic dehydrogenase(LDH) activity were determined, autophagic vesicles were counted, and the expression of YTHDF2, NRF2 and NCOA4 was detected by Western blot. Results:Part Ⅰ Animal experiment At the end of myocardial ischaemia-reperfusion, serum levels of CK-MB and cTnI and the percentage of myocardial infarct size were significantly increased, the expression of YTHDF2 and NCOA4 in myocardial tissues was up-regulated, and the expression of NRF2 was down-regulated in both diabetic and non-diabetic groups ( P<0.05). Compared with NIR group, serum levels of CK-MB and cTnI and the percentage of myocardial infarct size were significantly increased, the expression of YTHDF2 and NCOA4 in myocardial tissues was up-regulated, and the expression of NRF2 was down-regulated in DIR group ( P<0.05). Compared with DIR group, serum levels of CK-MB and cTnI and the percentage of myocardial infarct size were significantly decreased, the expression of YTHDF2 and NCOA4 in myocardial tissues was down-regulated, and the expression of NRF2 was up-regulated ( P<0.05), and the pathological damage was reduced in AAV-Y + DIR group. Part Ⅱ Cell experiment Compared with HG and HR groups, the cell viability was significantly decreased, the activity of LDH was increased, the counts of autophagic vesicle were increased, the expression of YTHDF2 and NCOA4 was up-regulated, and the expression of NRF2 was down-regulated in HHR group ( P<0.05). Compared with HHR group, the cell viability was significantly decreased, the activity of LDH was increased, the counts of autophagic vesicle were increased, the expression of YTHDF2 and NCOA4 was up-regulated, and the expression of NRF2 was down-regulated in M + HHR group, and the cell viability was significantly increased, the activity of LDH was decreased, the counts of autophagic vesicle were decreased, the expression of YTHDF2 and NCOA4 was down-regulated, and the expression of NRF2 was up-regulated in siY + HHR group ( P<0.05), and no statistically significant changes were found in the above indicators in siY + M + HHR group ( P>0.05) Conclusions:YTHDF2 can down-regulate the expression of NRF2, enhance the level of ferritinophagy, and participate in the process of MIRI in diabetic rats.