ObjectiveTo investigate the therapeutic effect of Astragali Radix-mediated changes in gut microbiota on treating type 2 diabetes （T2DM）. MethodsA 12-week randomized， placebo-controlled clinical trial enrolled eighty patients with newly diagnosed type 2 diabetes and poor glycemic control in the Qi deficiency type. All patients received insulin therapy. The observation group （40 cases） was administered with Astragali Radix Granules， while the control group （40 cases） received a placebo. Both treamtents were taken orally twice daily. Changes in gut microbiota were assessed by 16s rDNA sequencing. Serum glucagon-like peptide-1 （GLP-1） levels were measured using enzyme-linked immunosorbent assay （ELISA）. Glucose metabolism indicators including fasting blood glucose （FPG）， 2-hour postprandial blood glucose （2 h PG），glycated albumin（GA）， and glycated hemoglobin （HbA1c） were evaluated. Pancreatic function was evaluated using fasting C-peptide （FCP）， 2-hour postprandial C-peptide （2 h CP）， and C-peptide area under the curve （AUC_cp）. Traditional Chinese medicine （TCM） syndrome scores， clinical efficacy， and safety indicators were also observed. ResultsIn terms of glucose metabolism indicators， compared with the baseline， both groups exhibited significantly lower FPG， 2 h PG， GA and HbA1C （P<0.01），while FCP， 2 h CP and AUC_cp were significantly higher （P<0.01）. Compared with the control group after the treatment， the observation group showed significantly lower FPG， 2 h PG， GA and HbA1C（P<0.05， P<0.01），and significantly higher FCP， 2 h CP and AUC_cp （P<0.05， P<0.01）， indicating that Astragali Radix can improve glucose metabolism. In terms of the diversity of gut microbiota， no significant differences were detected in the Chao1， Shannon and Simpson indexes of the two groups compared with their respective baselines. However， compared with the post-treatment control group， the observation group demonstrated significant increases in the Chao1， Shannon and Simpson indexes （P<0.05， P<0.01）. The β-diversity analysis showed significant separation in gut microbiota composition before and after treatment in both groups， indicating that Astragali Radix can significantly alter the structure and improve the diversity of gut microbiota. At the phylum level， compared with the baseline， both groups showed a significant increase in the relative abundance of Bacteroidota（P<0.01）. The relative abundance of the potentially harmful phylum Proteobacteria was significantly lower in the observation Group after treatment （P<0.01）. Compared with the post-treatment control group， the observation group had a significantly higher relative abundance of Bacteroidota（P<0.01）. No significant difference was found in Firmicutes/Bacteroidota （F/B） ratio between the two groups after treatment， and other phyla showed no significant differences. At the genus level， compared with the baseline， the observation group exhibited a significant increase in Bacteroides （P<0.01） and a significant decrease in Escherichia-Shigella （P<0.01）， whereas no significant difference was seen in the control group . Compared with the control group after treatment， the observation group after treatment had a significantly higher relative abundance of Bacteroides （P<0.01）. No significant differences were seen in other genera. Linear discriminant analysis （LDA） identified potential characteristics taxa： in the observation group， Bacteroidota at the phylum level and Bacteroides and Dubosiella at the genus level， in the control group， Proteobacteria at the phylum level as well as Barnesiella and Staphylococcus at the genus level. Correlation analysis based on a heatmap revealed that GLP-1 levels were positively correlated with Firmicutes， F/B ratio and Fusobacterium， and negatively correlated with Bacteroidota， Proteobacteria， Bacteroides and Escherichia-Shigella. In terms of clinical efficacy， compared with the control group， the total effective rate of the observation group was significantly higher （P<0.05）. Compared with the baseline， the scores for shortness of breath， fatigue， weakness， spontaneous sweating and reluctance to speak significantly decreased in both groups （P<0.01）. Compared with the control group after treatment， the score for weakness was significantly lower in the observation group （P<0.01），indicating that Astragali Radix could improve clinical symptoms and alleviate weakness symptoms. In terms of safety， compared with the baseline， alanine aminotransferase （ALT） levels significantly decreased in both groups （P<0.05，P<0.01），indicating that Astragali Radix did not induce any significant abnormalities in liver and kidney functions. ConclusionAstragali Radix demonstrates the potential to significantly improve the gut microbiota environment in patients of newly diagnosed type 2 diabetes with Qi deficiency. The therapeutic effect may contribute to glycemic control， possibly mediated by an elevation in GLP-1 level. These findings may support its further clinical investigations and potential applications.

Objective To explore the construction of heart preservation model of empty beating donor based on extracorporeal membrane oxygenation (ECMO). Methods From January 2022 to August 2023, 20 Guangxi Bama miniature pigs weighing 25-30 kg were selected, half male and half female. Under general anesthesia and heparinization, a midline thoracotomy was performed. The pericardium was cut after freeing the anterior and posterior vena cavae, and a perfusion needle was inserted near the brachiocephalic artery in the ascending aorta, connected to a blood collection bag to collect 500-600 mL of blood. The anterior and posterior vena cavae were ligated, the aorta was blocked and perfused with HTK solution to stop the heart beating. The superior and inferior vena cavae were cut off, the right pulmonary vein was decompressed, the aorta and left and right pulmonary arteries and veins were cut off, and the whole heart was removed. An ECMO device was used to continuously perfuse a cardioprotective solution mainly composed of oxygenated warm blood, maintaining the isolated pig heart beating for 8 hours, monitoring (once/hour) ECMO perfusion parameters, blood gas indicators, perfusate electrolytes, inflammatory factors, myocardial enzymes, myoglobin, and troponin levels. Myocardial tissue was taken for hematoxylin-eosin (HE) staining to observe myocardial cell damage and evaluate the quality of heart preservation. Results Among the 20 isolated beating pig hearts, 17 successfully resumed beating, 3 experienced ventricular fibrillation, resuscitated after intracardiac electrical defibrillation, and all 20 pig hearts successfully beat for 8 hours. There was no statistical difference in ECMO perfusion parameters, blood gas indicators, perfusate electrolytes, and inflammatory factors at each time point (P>0.05). There were statistical increases in myocardial enzymes, myoglobin, and troponin levels (P<0.05). HE staining results suggested that there was no severe myocardial damage. Conclusion ECMO technology can be used for pig heart preservation with good results, and this study provides experimental evidence for improving heart preservation research in clinical heart transplantation.

Objectives: . This study aimed to explore the role of the Pcdh15 gene in hearing maintenance and to examine the effects of its deficiency on cochlear structure, cochlear function, and hearing loss in mice. Methods: . CRISPR/Cas9 technology was used to generate Pcdh15 knockout (KO) mice. Auditory function was evaluated using hearing tests, while histological approaches were employed to assess morphological changes in cochlear hair cells and spiral ganglion neurons (SGNs). RNA sequencing (RNA-seq) was performed on cochlear tissue from postnatal day 18 (P18) wild-type (WT) and Pcdh15 KO mice to identify differentially expressed genes (DEGs). These DEGs were subjected to functional annotation and pathway analysis, with particular emphasis on oxidative phosphorylation (OXPHOS). Reactive oxygen species (ROS) levels were quantified using flow cytometry and DCFH-DA assays. Results: . Pcdh15 KO mice exhibited progressive sensorineural hearing loss, which worsened to profound deafness by P18. Notably, cochlear hair cells and SGNs showed substantial loss and pronounced morphological abnormalities, particularly within the basal high-frequency region. RNA-seq analysis identified 1,359 DEGs, including 1,024 downregulated and 335 upregulated genes. Pathway analysis indicated that Pcdh15 deficiency was associated with inhibition of the OXPHOS pathway, potentially disrupting mitochondrial respiratory chain complexes I, III, IV, and V and thereby impairing energy production and ATP metabolism. In addition, early-stage cochleae from KO mice demonstrated elevated ROS levels and increased oxidative stress, suggesting that redox imbalance may contribute to hearing damage. Conclusion . Pcdh15 plays a critical role in hearing maintenance. Its deficiency is associated with severe hearing loss and marked cochlear abnormalities, which are linked to inhibition of OXPHOS, disruption of energy metabolism, and exacerbation of oxidative stress. Together, these findings provide new insights into the mechanisms underlying hearing loss and suggest potential targets for therapeutic intervention.

To evaluate patient satisfaction levels in a gynecological teaching clinic and to explore its associated influencing factors, so as to provide evidence for optimizing the operational model of teaching clinics.

Objective:To establish a novel in vivo and in vitro model of type 2 endoleaks (T2EL) after endovascular aneurysm repair (EVAR) of abdominal aortic aneurysm (AAA).Methods:In vitro model: Using 3Dmax (2022) software, models of AAA and T2EL were created. These models were fabricated using 3D printing technology, with quartz as the material. Fresh pig blood was injected into the models, and a circulation system was established using a peristaltic pump. The direction of blood flow was observed to confirm the establishment of the in vitro T2EL model. In vivo model: Six Bama miniature pigs were selected, each 10 months old and weighing 40-50 kg. After determining whether unilateral iliac artery occlusion could be permanent through experiments, an AAA animal model from the end of the abdominal aorta to the left common iliac artery was created using the patch formation method. EVAR was performed through the right femoral artery, completely occluding the left iliac artery. The T2EL model was established by using the retrograde blood flow from the left iliac artery. The formation of AAA and T2EL models in vivo was confirmed by color Doppler ultrasound, digital subtraction angiography (DSA), and other methods. The measurement data with normal distribution were expressed as mean±standard deviation( ± s), and comparisons between groups were performed using the independent-samples t-test. The comparison between groups of count data was conducted using the chi-square test. Results:The successful establishment of the T2EL model was confirmed by observing the blood flow in the circulating drip flask. The weight of the Bama miniature pigs in the in vivo model was (41.33±2.21) kg, the diameter of the abdominal aorta was (7.45±0.15) mm, and the maximum diameter of the aneurysm was (16.67±0.61) mm. Through experiments, it was determined that the left iliac artery and umbilical artery of the experimental pigs could be isolated. Therefore, after the aneurysm from the end of the abdominal aorta to the left common iliac artery was established, a covered stent was implanted from the abdominal aorta to the right common iliac artery for EVAR treatment. The successful establishment of the AAA model and the T2EL model with the left iliac artery as the main "culprit vessel" was confirmed by color Doppler ultrasound and DSA.Conclusion:The in vitro models established through 3D printing technology and the T2EL animal model with the left iliac artery as the main "culprit vessel" have a high success rate and can serve as a model basis for the mechanism and treatment research of T2EL after EVAR for AAA.

Objective To investigate the therapeutic mechanism of Dingkun Dan(DKD)in polycystic ovary syndrome(PCOS)by examining the effects of microRNA-30d-5p on ovarian granulosa cells(GCs).Methods A PCOS rat model was established using dehydroepiandrosterone(DHEA).Normal rat GCs and PCOS rat GCs were cultured in vitro and divided into four groups:blank control group,model group,low-dose DKD group,and high-dose DKD group.After grouping,GCs viability was assessed using the methyl thiazolyl tetrazolium(MTT)assay,GCs apoptosis was analyzed by flow cytometry,and the gene expression of transforming growth factor β1(TGF-β1),Smad2,Smad3,and microRNA-30d-5p in GCs was measured by real-time polymerase chain reaction(RT-PCR),protein expression of TGF-β1,Smad2,and Smad3 in GCs was detected by Western Blot.Results Compared with the blank control group,the model group exhibited significantly decreased GCs viability,increased GCs apoptosis,upregulated mRNA and protein expression of TGF-β1,Smad2,and Smad3,and downregulated microRNA-30d-5p expression,the differences were statistically significant(P＜0.01).Compared with the model group,both low-and high-dose DKD groups showed increased GCs viability,reduced GCs apoptosis,downregulated mRNA and protein levels of TGF-β1 Smad2 and Smad3,elevated microRNA-30d-5p expression,and the differences were statistically significant(P＜0.05 or P＜0.01).Conclusion DKD promotes GCs proliferation by targeting Smad2 via microRNA-30d-5p,suggesting a potential therapeutic role in PCOS-related ovulatory dysfunction.

Objective:To investigate the predictive value of microRNA-1(miR-1),brain natriuretic peptide(BNP)and ischemia-modified albumin(IMA)for unstable angina pectoris(UAP).Methods:We enrolled 237 UAP patients admitted to Xining Second People's Hospital between June 2018 and December 2020 as UAP group.Another 86 healthy subjects undergoing physical examination simultaneously were enrolled as control group.MiR-1 expression,BNP and IMA levels were measured.General data between UAP group and control group,serum miR-1 expression,BNP and IMA levels among different Braunwald class and prognosis were compared.Receiver operat-ing characteristic(ROC)curve was employed to analyze predictive value of miR-1,BNP,IMA and their combina-tion for prognosis in UAP patients.Results:Compared with participants in the control group,those in UAP group had significant higher serum miR-1 expression[(1.80±0.59)vs.(0.93±0.11)],BNP[(107.34±37.46)pg/ml vs.(52.31±10.64)pg/ml]and IMA[(79.76±19.29)g/L vs.(53.16±6.43)g/L](P＜0.001 all).As Braun-wald class increased(class Ⅰ～Ⅲ),serum miR-1 expression,BNP and IMA levels elevated(P＜0.001 all).Com-pared with patients in favorable outcome group,those in unfavorable outcome group had significant higher serum miR-1 expression[(2.31±0.54)vs.(1.53±0.41)],BNP[(147.03±29.63)pg/ml vs.(85.95±19.46)pg/ml]and IMA[(97.24±15.35)g/L vs.(70.35±13.88)g/L](P＜0.001 all).ROC curve indicated that AUC of com-bined detection for predicting unfavorable outcome in UAP patients was 0.925(95％CI 0.884～0.955),which was significantly higher than miR-1(AUC=0.880,95％CI 0.831～0.918),BNP(AUC=0.863,95％CI 0.813～0.904)and IMA(AUC=0.900,95％CI 0.854～0.935)alone(Z=2.884,3.130,2.090,P＜0.05 or＜0.01).Conclusion:MiR-1 expression,BNP and IMA levels significantly increase in UAP patients,and they are associated with the severity of disease.Combined detection has good predictive value for unfavorable outcome in UAP pa-tients.

Objective:To explore the application effect of ADDIE model (analysis, design, development, implementation and evaluation) in the training of maxillofacial trauma first-aid ability of dental nurses, in order to optimize the training process of dental nurses.Methods:A self-controlled before and after study was conducted. Fifty-one dental nurses in Qingdao Stomatology Hospital Affiliated to Qingda University were selected as the research objects by convenient sampling method in March 2022, and the first aid ability training for maxillofacial trauma was carried out according to the five stages of ADDIE model. Before and after the training, the evaluation was made from three aspects: theory and skill test results, first-aid ability and training satisfaction scores.Results:Among 51 dental nurses, there were 4 males and 47 females, aged (30.69 ± 5.85) years. After the training, the theoretical assessment score of dental nursing staff was (88.87 ± 6.20) points, higher than that before the training (80.51 ± 7.21) points, and the technical assessment score was (91.61 ± 4.08) points, higher than that before the training (82.03 ± 7.56) points. The differences were statistically significant ( t = - 14.38, - 10.93, both P<0.01). The total score of first aid ability of nurses was (137.38 ± 11.30) points, higher than that before training (123.40 ± 13.73) points, and the difference was statistically significant ( t = - 17.30, P<0.01). The satisfaction score of dental nursing staff was (4.58 ± 0.50) points, higher than that before training (3.96 ± 0.46) points, and the difference was statistically significant ( t = - 11.51, P<0.01). Conclusions:The training program based on ADDIE model, through systematic teaching design, is helpful to improve the emergency treatment ability and training satisfaction of dental nursing staff with maxillofacial trauma.

Objective To explore the incidence and its influencing factors of transient severe motion(TSM)in the arterial phase of gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid(Gd-EOB-DTPA)enhanced MRI.Methods The databases of China National Knowledge Network(CNKI),VIP,Wanfang,PubMed,and Embase were searched for studies on the incidence and influencing factors of TSM,and the search time was from the establishment of the databases to October 2024.Meta-analysis was performed via Stata 17.0 software.Results A total of 30 papers(33 studies)were finally included,totaling 12 565 patients.Meta-analysis results showed that the incidence of TSM in the arterial phase of Gd-EOB-DTPA enhanced MRI was 13.0％.The risk factors for TSM included age[odds ratio(OR)=1.03;95％confidence interval(CI)1.02-1.05;P＜0.001),chronic obstructive pulmonary disease(COPD)(OR=4.21;95％CI 1.76-10.09;P=0.001),and moderate-to-severe pleural effusion(OR=3.34;95％CI 1.69-6.63;P=0.001),while a previous usage history of Gd-EOB-DTPA(OR=0.56;95％CI 0.39-0.81;P=0.002)was a protective factor of TSM.Conclusion The incidence of TSM in the arterial phase of Gd-EOB-DTPA enhanced MRI is relatively high.Age,COPD,moderate-to-severe pleural effusion are risk factors for TSM,while the previous usage history of Gd-EOB-DTPA is a protective factor for TSM.

Lentiviral vectors(LVs)are key gene delivery tools for integrating target genes into the host genome,but they may also pose risks of insertional mutagenesis.The vector copy number(VCN)in cells is critical for determining the safety of gene modification.However,the reliability and accuracy of its quantification process are influenced by multiple factors.Developing cell reference materials with specific vector copy numbers represents a viable approach to enhance the reliability and consistency of measurement results,enabling quality control of the quantification process and traceability of outcomes.However,the preparation of such reference materials faces challenges in cell sample design,preparation protocols,and advanced quantification techniques.In this study,T lymphocyte cell line Jurkat-based reference materials with LV gene copy numbers of 1 and 2 copy/cell were developed.A high-precision duplex digital polymerase chain reaction(dPCR)method was established to quantify the LV gene and endogenous genes simultaneously.Additionally,the results of dPCR were cross-validated through next-generation sequencing and flow cytometric analysis.Ultimately,confocal microscopy characterization results showed that the developed cell reference materials had intact morphology.The quantification result of VCN-1 was(1.07±0.11)copy/cell,and that of VCN-2 was(2.09±0.21)copy/cell.These cell reference materials demonstrated compliance with stability and homogeneity requirements,and could be applied for quality control throughout the VCN measurement workflow and metrological traceability,improving the accuracy,comparability,and validity of copy number measurements.