ObjectiveTo investigate the mechanism by which Notoginsenoside R1 （NGR1） ameliorates hypoxia/reoxygenation （H/R）-induced injury in AC16 human cardiomyocyte cell lines through the regulation of mitophagy. MethodsCommon genes linked to hypoxia/reoxygenation injury and mitophagy were identified by intersecting data from GeneCards and MitoCarta databases. AC16 cell viability was assessed via CCK-8 assay under varying NGR1 concentrations （0， 6.25， 12.5， 25， 50， 100， 200， 300， 400， 500 μmol/L）. AC16 cells were divided into the following groups： control group （Control）， model group （H/R）， and treatment groups （H/R + NGR1 at 100， 200 and 300 μmol/L）. Mitochondrial membrane potential （ΔΨm） was measured using 5，5'，6，6'-tetrachloro-1，1'，3，3'-tetraethylbenzimidazolylcarbocyanine iodide （JC-1） staining. Transcriptional levels of mitophagy-related genes （Parkin， Pink1， P62） were quantified by reverse transcription-quantitative PCR （RT-qPCR）. Protein expression of mitophagy-related markers （Parkin， Pink1， P62， and LC3BⅡ） was evaluated via Western blot analysis. Mitochondrial ultrastructure was visualized by transmission electron microscopy （TEM）. ResultsCompared to the control group， cell viability in the H/R group significantly decreased （P<0.01）. Treatment with NGR1 at concentrations above 100 μmol/L significantly enhanced the cell viability of AC16 cells compared to the H/R group （P<0.01）. H/R induced a significant decrease in mitochondrial membrane potential （P<0.01）， which was restored by NGR1 treatment （P<0.01）. The mRNA levels of Parkin， Pink1， and P62 in the H/R group were upregulated compared to the control group （P<0.05）， while NGR1 intervention downregulated their expression （P<0.05）. Protein expression levels of Parkin， Pink1， and LC3BⅡ in the H/R group significantly increased， while P62 expression decreased compared to the control group （P<0.01）. In contrast， different doses of NGR1 treatment significantly reduced the expression of Parkin， Pink1， and LC3BⅡ while increasing P62 expression （P<0.05）. TEM revealed that the mitochondrial structure in the H/R group was severely disrupted， with fragmented and disorganized cristae， which was alleviated by NGR1. ConclusionNGR1 ameliorates H/R-induced AC16 cell injury， and its mechanism may be associated with modulating the Pink1/Parkin pathway to suppress excessive mitophagy.

ObjectiveThe U6 promoter is an essential element for driving sgRNA expression in the clustered regularly interspaced short palindromic repeat sequences/CRISPR-associated protein 9（CRISPR/Cas9）gene editing system in dicotyledonous plants. Endogenous U6 promoters typically exhibit higher transcriptional activity， which can significantly improve gene editing efficiency. This study aims to identify endogenous U6 promoters in Artemisia annua to optimize its CRISPR/Cas9 gene editing system， which holds significant importance for its molecular breeding. MethodsOn the basis of the highly conserved U6 snRNA sequences in Arabidopsis thaliana， endogenous U6 promoters were screened in the A. annua genome. Expression vectors were constructed with candidate AaU6 promoter driving the firefly luciferase （LUC） reporter gene， and then transiently transformed into Nicotiana benthamiana. Transcriptional activities of the promoters were measured and compared by in vivo imaging and the Dual Luciferase Reporter assay. ResultsEight endogenous U6 promoters were successfully cloned from A. annua. Sequences alignment revealed that all these promoters contained the two conserved cis-acting elements， upstream sequence element （USE） and TATA-box， which affected their transcriptional activity. Dual-luciferase activity assays indicated that the transcriptional activities of AaU6-3， AaU6-1， and AaU6-5 were significantly higher than that of the Arabidopsis AtU6-26 promoter， with AaU6-3 exhibiting the highest activity. ConclusionThis study identified three endogenous AaU6 promoters with high transcriptional activity in A. annua， providing key functional elements for establishing an efficient gene editing system in A. annua. These findings will contribute to advancing precision molecular breeding and high-quality germplasm innovation in A. annua.

Chemotherapy is currently the mainstay of systemic management for triple-negative breast cancer (TNBC), but chemoresistance significantly impacts patient outcomes. Our research indicates that Doxorubicin (Dox)-resistant TNBC cells exhibit increased glycolysis and ATP generation compared to their parental cells, with this metabolic shift contributing to chemoresistance. We discovered that ALKBH3, an m¹A demethylase enzyme, is crucial in regulating the enhanced glycolysis in Dox-resistant TNBC cells. Knocking down ALKBH3 reduced ATP generation, glucose consumption, and lactate production, implicating its involvement in mediating glycolysis. Further investigation revealed that aldolase A (ALDOA), a key enzyme in glycolysis, is a downstream target of ALKBH3. ALKBH3 regulates ALDOA mRNA stability through m¹A demethylation at the 3'-untranslated region (3'UTR). This methylation negatively affects ALDOA mRNA stability by recruiting the YTHDF2/PAN2-PAN3 complex, leading to mRNA degradation. The ALKBH3/ALDOA axis promotes Dox resistance both in vitro and in vivo. Clinical analysis demonstrated that ALKBH3 and ALDOA are upregulated in breast cancer tissues, and higher expression of these proteins is associated with reduced overall survival in TNBC patients. Our study highlights the role of the ALKBH3/ALDOA axis in contributing to Dox resistance in TNBC cells through regulation of ALDOA mRNA stability and glycolysis.

There is a large gap between production and demand of plant oil in China, which leads to the heavy reliance on imports. Diacylglycerol acyltransferase (DGAT) and phospholipid: diacylglycerol acyltransferase (PDAT) are two key enzymes responsible for the synthesis of triacylglycerol, thereby affecting the yield and quality of plant oil. This paper comprehensively reviews the research progress in DGAT and PDAT in terms of their biological functions in plant oil synthesis, the molecular mechanisms of regulating plant lipid metabolism, growth, and development under stress, and their roles in driving oil synthesis under the background of synthetic biology. Furthermore, future research and application of DGAT and PDAT are prospected. This review aims to provide a basis for deeply understanding the molecular mechanism of plant oil synthesis and improving the quality and productivity of oil crops by the utilization of DGAT and PDAT genes.
Diacylglycerol O-Acyltransferase/physiology* ; Plant Oils/metabolism* ; Acyltransferases/metabolism* ; Lipid Metabolism/genetics* ; Gene Expression Regulation, Plant ; Triglycerides/biosynthesis*

The occurrence and development of tumors are determined by both intrinsic and microenvironmental factors. The components of the tumor microenvironment exhibit great heterogeneity resulting in either pro-tumor or anti-tumor effects. The deep exploration of tumor immune microenvironment gave birth to the hypothesis of cancer immunoediting, which is supported by more and more evidence. Cancer immunoediting is accomplished through the interactions among various cells and factors in the microenvironment. While many molecular mechanisms have been studied, the cellular mechanisms of cancer immunoediting have been paid attention. This article explores the roles of cellular mechanisms in the cancer immunoediting, including immunogenic cell death, cellular senescence and neutrophil extracellular trap. The elucidation of these mechanisms not only reveals the complexity of the pathogenesis and progression of tumors, but also provides new targets and prognostic indicators for the diagnosis and treatment of tumors.

Objective·To evaluate the relationships between disc-condyle distance and anterior disc displacement,as well as between disc-condyle distance and disc morphology,in patients with temporomandibular disorders(TMD)using magnetic resonance imaging(MRI)of the temporomandibular joint(TMJ).Methods·From September 2023 to March 2024,90 patients(180 TMJs)who visited the TMJ clinic of Department of Stomatology,The Second Affiliated Hospital of Anhui Medical University,with clinical symptoms of TMD and were diagnosed via MRI with either anterior disc displacement or no significant displacement,were included.Clinical data were collected,and MRI images were used to measure the angle of disc displacement,disc-condyle distance,disc length,and thickness.The degree of disc deformation was assessed.The relationships between clinical symptoms and anterior disc displacement,between anterior disc displacement and both disc morphology and disc-condyle distance,and between disc-condyle distance and disc morphology were analyzed.Results·Among the 90 patients,there were 16 males and 74 females,with a mean age of(28.1±14.5)years.Among the 180 TMJs,175 had clinical symptoms and 5 were asymptomatic.There were 40 joints with no displacement,78 with reducible anterior disc displacement,and 62 with irreducible anterior disc displacement.In the joints with irreducible anterior disc displacement,the proportion of those with two or more symptoms was slightly higher at 62.9%,but the difference was not statistically significant compared with the joints with no displacement or reducible anterior disc displacement.MRI assessment revealed that in the joints with irreducible anterior disc displacement,the proportion of disc deformation type Ⅲ or higher was significantly higher compared with the non-displaced joints(P<0.001).The disc length was significantly shorter(P<0.001),and the intermediate zone thickness was significantly greater(P<0.001)compared with the non-displaced joints.The disc displacement angles at centric closure and maximum opening were also significantly larger(P<0.001).The disc-condyle distance was 3.10(2.70,3.70)mm for non-displaced joints,3.40(3.00,4.00)mm for joints with reducible anterior disc displacement,and 6.60(4.78,7.90)mm for joints with irreducible anterior disc displacement,with significant differences(P<0.001).The disc-condyle distance was 3.10(2.80,3.60)mm for type Ⅰ discs,3.70(3.10,4.60)mm for type Ⅱ discs,5.10(4.00,7.30)mm for type Ⅲ discs,and 6.80(4.98,8.20)mm for type Ⅳ/Ⅴ discs,with significant differences(P<0.001).The disc-condyle distance was negatively correlated with disc length(rs=-0.469,P<0.001),positively correlated with intermediate zone thickness(rs=0.319,P<0.001),and positively correlated with disc displacement angle at centric closure(rs=0.626,P<0.001).Conclusion·With increasing severity of disc deformation,intermediate zone thickness,and disc displacement angle at centric closure,as well as decreasing disc length,the disc-condyle distance increases.The disc-condyle distance is an important indicator for MRI assessment of pathological changes in TMD.

Objective To explore the effectiveness of integrating virtual simulation technology with case-based learning(CBL)in the teaching of clinical blood transfusion testing technology.Methods From January to June 2024,99 undergraduate students majoring in medical laboratory science at our university were selected and divided into an observation group(50 students)and a control group(49 students)based on their classes.The control group followed a traditional theoretical teaching model,while the observation group employed virtual simulation technology combined with CBL.Results The study compared as-sessment scores for theory learning,practical skills,case analysis,and comprehensive quality,teaching effects such as coordina-tion and cooperation abilities,self-efficacy measured by the GSES scale,and teaching satisfaction between the two groups.The results indicated that the observation group had significantly higher scores in all assessment categories,teaching effect abilities,and total GSES scores compared to the control group(P＜0.05).The total teaching satisfaction rate in the observation group reached 100%,significantly higher than the 80% in the control group(P＜0.05).Conclusion The integration of virtual simu-lation technology with CBL innovates the teaching model for clinical blood transfusion testing courses,effectively enhancing students'professional skills and comprehensive quality,boosting self-efficacy,and achieving high student satisfaction,demon-strating significant potential for broader application.

Objective To explore the relationship between hemorheology and Carotid Atherosclerosis.Methods The clinical data of 153 patients who underwent both hemorheological testing and carotid artery ultrasound were divided into a CAS group(n=96)and a non-CAS group(n=57)based on ultrasound findings.Clinical data and laboratory indicators were compared between two groups.Multivariate logistic regression analysis was used to explore the influencing factors of CAS.The ROC curves graph were drawn to observe the role of hemorheological indicators in predicting CAS and select the optimal cutoff value based on the maximum Youden index.Results The CAS group demonstrated higher levels in age,BMI,RBC aggregation index,low&high shear reduced viscosity of whole blood,plasma viscosity and fibrinogen compared to the non-CAS group(P<0.05).The multivariate logistic regression analysis showed that plasma viscosity(OR=38.270,95%CI:1.206～1214.508),age(OR=1.119,95%CI:1.065～1.176)were risk factors for the occurrence of CAS(P<0.05).The ROC curves showed that the area under the curve(AUC)of plasma viscosity and age were 0.623、0.728.Conclusion CAS patients have high levels of plasma viscosity and advanged age compared to the patient without CAS.Elevated plasma viscosity and age is a risk factor for CAS,with plasma viscosity≥1.46 mPa·s,over the age of 56.5 as a significant value for predicting CAS.

Purpose To investigate the value of intratumoral and different ranges of peritumoral radiomics features of the primary lesion of breast cancer based on ultrasound images in predicting axillary lymph node metastasis(ALNM),and to explore the best peritumoral range.Materials and Methods A total of 312 cases confirmed by pathology in breast cancer patients with preoperative ultrasound images from June 2022 to February 2024 in Binzhou Medical University Hospital were retrospectively enrolled,and were randomly divided into training set and testing set according to the 7∶3 proportion.The tumor border of the ultrasound images was manually delineated as the intratumoral region of interest,and the peritumoral region of interest was obtained by conformal automatically extended different range(1,2,3,4 and 5 mm).The radiomics features were screened.Based on the selected optimal radiomics features,random forest classifier was used to construct three types of radiomics models(intratumoral model,5 peritumoral models,and 5 intratumoral+peritumoral models).The performance and clinical practicability of the models was assessed the area under the curve(AUC)and decision curve analysis.Results The AUCs of the intratumoral+peritumoral radiomics models for predicting ALNM in the training set and test set were 0.807-0.873,0.728-0.780,respectively,which were superior to those of the single intratumoral radiomics models(0.822,0.758)and peritumoral radiomics models(0.722-0.768,0.650-0.710).The intratumoral+peritumoral 3 mm radiomics model showed the best predictive performance,with AUC of 0.873 in the training set and 0.780 in the test set,respectively,and the decision curve showed that the model had a good clinical net benefit.Conclusion The combined intratumoral and peritumoral radiomics features of the primary lesion of breast cancer based on ultrasound images can effectively predict ALNM,and 3 mm peritumoral may be the best peritumoral range for predicting ALNM.