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.

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.

Objective:To compare the effects of propofol and dexmedetomidine for sedation on the outcome in mechanically ventilated patients with severe pulmonary infection.Methods:Patients with severe pulmonary infection (Clinical Pulmonary Infection Score >7) requiring mechanical ventilation from the Medical Information Mart for Intensive Care Ⅳ database between 2008 and 2020 were selected and divided into propofol group and dexmedetomidine group based on the sedative agent used. The primary outcome was all-cause in-hospital mortality, and secondary outcomes included 90-day all-cause mortality and duration of mechanical ventilation. Inverse probability of treatment weighting (IPTW) was used to adjust for baseline confounders, and logistic regression and linear regression were applied to analyze the effects of the two sedative drugs on the outcomes of mechanically ventilated patients with pulmonary infection. Kaplan-Meier survival curves were used to analyze survival outcomes.Results:A total of 6 204 critically ill patients with pulmonary infection requiring mechanical ventilation were included, with 3 439 cases in propofol group and 2 765 cases in dexmedetomidine group. The baseline characteristics between the two groups were well balanced (standardized mean difference < 0.1) after IPTW adjustment. In the IPTW-adjusted cohort, the in-hospital all-cause mortality and 90-day all-cause mortality were significantly lower in dexmedetomidine group than in propofol group (439.2[18.7%] vs 563.6[24.1%], 618.0[26.3%] vs 733.6[31.3%], P<0.001), the results of Further Kaplan-Meier survival curve analysis showed that the 90-day all-cause mortality was significantly lower in dexmedetomidine group than in propofol group ( P<0.01). Conclusions:Compared with propofol, dexmedetomidine can decrease the mortality rate and improve the prognosis in mechanically ventilated patients with severe pulmonary infection when used for sedation.

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.

T-cell acute lymphoblastic leukemia (T-ALL) is a highly aggressive hematologic malignancy with a poor prognosis, despite advancements in treatment. Many patients struggle with relapse or refractory disease. Investigating the role of the super-enhancer (SE) regulated gene ubiquitin-specific protease 20 (USP20) in T-ALL could enhance targeted therapies and improve clinical outcomes. Analysis of histone H3 lysine 27 acetylation (H3K27ac) chromatin immunoprecipitation sequencing (ChIP-seq) data from six T-ALL cell lines and seven pediatric samples identified USP20 as an SE-regulated driver gene. Utilizing the Cancer Cell Line Encyclopedia (CCLE) and BloodSpot databases, it was found that USP20 is specifically highly expressed in T-ALL. Knocking down USP20 with short hairpin RNA (shRNA) increased apoptosis and inhibited proliferation in T-ALL cells. In vivo studies showed that USP20 knockdown reduced tumor growth and improved survival. The USP20 inhibitor GSK2643943A demonstrated similar anti-tumor effects. Mass spectrometry, RNA-Seq, and immunoprecipitation revealed that USP20 interacted with hypoxia-inducible factor 1 subunit alpha (HIF1A) and stabilized it by deubiquitination. Cleavage under targets and tagmentation (CUT&Tag) results indicated that USP20 co-localized with HIF1A, jointly modulating target genes in T-ALL. This study identifies USP20 as a therapeutic target in T-ALL and suggests GSK2643943A as a potential treatment strategy.

Astragalus membranaceus possesses the function of enhancing immunity, protecting the liver, diuretic, anti-aging, anti-stress, anti-hypertensive, and more extensive antibacterial effects. Polysaccharides, one kind of the major active ingredients of A. membranaceus, are considered to be responsible for their versatile use. Now, a systematic summary of research progress and prospects of polysaccharides from A. membranaceus polysaccharides (AMPs) is necessary to facilitate their further study and application. In this review, the optimal extraction methods, structural features, biological activities, and applications of AMPs were emphasized. The structure-activity relationships are also analyzed and elucidated. Solvent, ultrasonic, microwave, enzyme-assisted, ultra-high pressure, and combined methods have been used to extract AMPs. Among them, solvent extraction is the most commonly used method because it is simple and easy to operate, but the efficiency needs to be improved further. The ultra-high pressure method is the most efficient but has a low economic return. AMPs exhibited various bioactivities, including immunomodulation, antitumor, and antidiabete. The structure-activity relationships revealed that different structure configurations, chain conformations, and physical properties would have different bioactivities. However, the new method for purification of certain polysaccharides, detailed structure-activity relationships (SAR), mechanisms of bioactivities, and quality control of AMPs need to be extensively investigated.

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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 epidemiological characteristics of respiratory human adenovirus(H AdV)infections in hospitalized children in Ningbo,and to provide data for formulating infection prevention and control strategies for HAdV.METHODS A total of 65 022 children hospitalized with respiratory infections at the Women and Children's Hospital of Ningbo University from Jul.2019 to Dec.2024 were selected as the study sub-jects.Multiple reverse transcription-polymerase chain reaction(RT-PCR)and capillary electrophoresis were used to detect 11 non-bacterial pathogens.Basic and clinical information of the children was collected for analysis.RESULTS A total of 65 022 specimens were tested,with 4 292 cases tested positive for HAdV positivity,yielding a positive rate of 6.60％.The lowest positive rate was observed in 2023(3.22％),while the highest was in 2024(13.97％).Compared to the years 2019-2023,the overall HAdV positive rate was high in 2024,peaking at 26.80％in Jun.,indicating an outbreak.The total HAdV positive rate was higher in boys(6.82％)than in girls(6.32％)(P=0.006).The highest HAdV positive rate was observed in the 2 to＜6 age group(9.00％),while the lowest was in the 0 to＜1 age group(2.33％).Among the HAdV-positive specimens,2 658 cases(61.93％)were single infections,and 1 634 cases(38.07％)were co-infections.The non-bacterial respiratory pathogens with the highest co-infection rates were human rhinovirus(34.09％),Mycoplasma pneumoniae(20.44％)and human parainfluenza virus(5.75％).CONCLUSIONS A n outbreak of HAdV infections is observed among hospitalized children in Ningbo in 2024.Higher positive rates are found in boys aged 2 to＜6 years,with a certain proportion of co-infections.