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.

Sanqi is first recorded in the Compendium of Materia Medica(Ben Cao Gang Mu) in the Ming Dynasty. During the Ming and Qing Dynasties, Sanqi, as a precious Dao-di herb, was successively spread and introduced for cultivation. This study verified the germplasm resources, production areas, and spread of Sanqi in the Ming and Qing Dynasties by systematically reviewing the historical materials, such as materia medica works and local chronicles, and the modern distribution of production areas. In the Ming and Qing Dynasties, the original plants of Sanqi included Panax notoginseng, P. japonicus, P. bipinnatifidus, P. zingiberensis, P. stipuleanatus, and Gynura japonica. Among them, the production area of P. notoginseng has changed. From 1578 to 1593, the main production areas of P. notoginseng were Nandan county, Hechi city in Guangxi Zhuang autonomous region and Guangnan county and Funing county, Wenshan prefecture in Yunnan province. From 1683 to 1755, the production areas of P. notoginseng additionally included Yizhou district, Tian'e county, and Huanjiang county in Hechi city, and Tianyang district and Tiandong county in Baise city, Xincheng county and Gongcheng county in Guangxi Zhuang autonomous region. From 1765 to 1892, the production areas additionally included Youjiang district, Debao county, Napo county, and Jingxi city in Baise city, and Tiandeng county in Guangxi Zhuang autonomous region, and Wenshan city, Malipo county, Yanshan county, Xichou county, and Maguan county in Wenshan prefecture, and Baoshan city, Dali prefecture, Lincang city, Honghe prefecture, Mangshi city, and Lushui city in Yunnan province. During the Wanli period of the Ming Dynasty, Sanqi was introduced to Zhejiang province. During the Qianlong period of the Qing Dynasty, it was introduced to Fujian province. During the Daoguang period of the Qing Dynasty, it was introduced to Hunan province. By comprehensively reviewing the materia medica works, local chronicles, and novel historical materials, this study restores the development history of the Sanqi industry in the Ming and Qing Dynasties. Historical data show that the introduction of Dao-di herbs should consider the biological characteristics of medicinal plants and avoid blind introduction.
China ; Drugs, Chinese Herbal/history* ; History, 17th Century ; History, 16th Century ; Plants, Medicinal/chemistry* ; Medicine, Chinese Traditional/history* ; History, 18th Century

Emodin is a hydroxyanthraquinone compound that is widely distributed and has multiple pharmacological activities, including anti-diarrheal, anti-inflammatory, and liver-protective effects. Research indicates that emodin may be one of the main components responsible for inducing hepatotoxicity. However, studies on the mechanisms of liver injury are relatively limited, particularly those related to bile acids(BAs) metabolism. This study aims to systematically investigate the effects of different dosages of emodin on BAs metabolism, providing a basis for the safe clinical use of traditional Chinese medicine(TCM)containing emodin. First, this study evaluated the safety of repeated administration of different dosages of emodin over a 5-week period, with a particular focus on its impact on the liver. Next, the composition and content of BAs in serum and liver were analyzed. Subsequently, qRT-PCR was used to detect the mRNA expression of nuclear receptors and transporters related to BAs metabolism. The results showed that 1 g·kg~(-1) emodin induced hepatic damage, with bile duct hyperplasia as the primary pathological manifestation. It significantly increased the levels of various BAs in the serum and primary BAs(including taurine-conjugated and free BAs) in the liver. Additionally, it downregulated the mRNA expression of farnesoid X receptor(FXR), retinoid X receptor(RXR), and sodium taurocholate cotransporting polypeptide(NTCP), and upregulated the mRNA expression of cholesterol 7α-hydroxylase(CYP7A1) in the liver. Although 0.01 g·kg~(-1) and 0.03 g·kg~(-1) emodin did not induce obvious liver injury, they significantly increased the level of taurine-conjugated BAs in the liver, suggesting a potential interference with BAs homeostasis. In conclusion, 1 g·kg~(-1) emodin may promote the production of primary BAs in the liver by affecting the FXR-RXR-CYP7A1 pathway, inhibit NTCP expression, and reduce BA reabsorption in the liver, resulting in BA accumulation in the peripheral blood. This disruption of BA homeostasis leads to liver injury. Even doses of emodin close to the clinical dose can also have a certain effect on the homeostasis of BAs. Therefore, when using traditional Chinese medicine or formulas containing emodin in clinical practice, it is necessary to regularly monitor liver function indicators and closely monitor the risk of drug-induced liver injury.
Emodin/administration & dosage* ; Bile Acids and Salts/metabolism* ; Animals ; Male ; Liver/injuries* ; Chemical and Drug Induced Liver Injury/genetics* ; Drugs, Chinese Herbal/adverse effects* ; Humans ; Rats, Sprague-Dawley ; Mice ; Rats

Photodynamic immunotherapy is a promising strategy for cancer treatment. However, the dysfunctional tumor vasculature results in tumor hypoxia and the low efficiency of drug delivery, which in turn restricts the anticancer effect of photodynamic immunotherapy. In this study, we designed photosensitive lipid nanoparticles. The synthesized PFBT@Rox Lip nanoparticles could produce type I/II reactive oxygen species (ROS) by electron or energy transfer through PFBT under light irradiation. Moreover, this nanosystem could alleviate tumor hypoxia and promote vascular normalization through Roxadustat. Upon irradiation with white light, the ROS produced by PFBT@Rox Lip nanoparticles in situ dysregulated calcium homeostasis and triggered endoplasmic reticulum stress, which further promoted the release of damage-associated molecular patterns, enhanced antigen presentation, and stimulated an effective adaptive immune response, ultimately priming the tumor microenvironment (TME) together with the hypoxia alleviation and vessel normalization by Roxadustat. Indeed, in vivo results indicated that PFBT@Rox Lip nanoparticles promoted M1 polarization of tumor-associated macrophages, recruited more natural killer cells, and augmented infiltration of T cells, thereby leading to efficient photodynamic immunotherapy and potentiating the anti-primary and metastatic tumor efficacy of PD-1 antibody. Collectively, photodynamic immunotherapy with PFBT@Rox Lip nanoparticles efficiently program TME through the induction of immunogenicity and oxygenation, and effectively suppress tumor growth through immunogenic cell death and enhanced anti-tumor immunity.

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.

Objective To investigate the correlation of serum brain natriuretic peptide(BNP)and D-dimer levels with cardiac function in elderly patients with chronic heart failure(CHF).Methods A retrospective analysis was conducted on 91 elderly CHF patients(observation group)admitted in our hospital from January 2021 to November 2024.They were then classified into grade Ⅰ-Ⅱsubgroup(66 cases)and grade Ⅲ-Ⅳ subgroup 25 cases according to NYHA cardiac function grading.Another 84 healthy individuals who underwent physical examinations in our hospital dur-ing the same period were selected as the control group.The levels of serum BNP and D-dimer were detected.Multivariate logistic regression model was used to analyze the risk factors affecting the severity of cardiac function in elderly CHF patients,and Spearman correlation analysis was adopted to analyze the correlation between the levels and cardiac function in elderly patients with CHF.Results Serum D-dimer and BNP levels were significantly higher in the observation group than the control group(P＜0.01),and in the grade Ⅲ-Ⅳ subgroup than the grade Ⅰ-Ⅱ sub-group(P＜0.01).Multivariate logistic regression analysis revealed that age(OR=2.815,95％CI:1.501-5.281,P=0.001),BNP(OR=2.901,95％CI:1.458-5.772,P=0.003)and D-dimer(OR=2.872,95％CI:1.501-5.495,P=0.002)were independent influencing factors for cardiac function in elderly CHF patients(P＜0.01).Spearman correlation analysis showed that serum BNP and D-dimer were negatively correlated with cardiac function(r=-0.324,P=0.023;r=-0.285,P=0.035).Conclusion The serum levels of D-dimer and BNP are significantly increased in elderly CHF patients,and the two levels are related to cardiac function.

Objective:To explore the risk factors for in-hospital death after complete recanalization by mechanical thrombectomy and establish a risk prediction model in patients with acute large vessel occlusion stroke of the anterior circulation.Methods:A total of 468 patients with anterior circulation acute large vessel occlusion stroke who underwent mechanical thrombectomy in Stroke Center (Second Affiliated Hospital of Dalian Medical University), Department of Interventional Therapy (First Affiliated Hospital of Dalian Medical University), and Department of Neurointervention and Neurocritical Care (Central Hospital Affiliated to Dalian University of Technology) from January 2016 to November 2023 were selected. All patients achieved complete recanalization (modified thrombolysis in cerebral infarction: grading 3) immediately after thrombectomy. The clinical data, laboratory and imaging results of the patients were collected, and these patients were divided into in-hospital death group ( n=52) and in-hospital survival group ( n=416) according to occurrence of in-hospital death (all-cause death). Univariate analysis and multivariate Logistic regression analysis were used to screen the risk factors for in-hospital death, and a risk prediction model was constructed. Receiver operating characteristic (ROC) curve was used to evaluate the predictive efficacy of the model, calibration curve and Hosmer-Lemeshow test were used to evaluate the accuracy of the model, and decision curve was used to evaluate the clinical utility of the model. Results:Univariate analysis showed that the in-hospital death group had significantly higher proportions of female patients, patients with atrial fibrillation, and patients with symptomatic intracranial hemorrhage compared with the in-hospital survival group (50.0% vs. 31.3%; 57.7% vs. 41.6%; 38.5% vs.11.8%), and significantly higher baseline blood glucose, and National Institutes of Health Stroke Scale score, neutrophil count, and neutrophil/lymphocyte ratio within 24 hours of thrombectomy (8.10 [7.05, 11.79] vs. 7.31[6.46, 9.25], 20 [16, 32] vs. 15 [10, 22], 10.09 [7.87, 13.19] vs. 8.47 [6.73, 10.32], 10.63 [5.87, 15.69] vs. 7.13 [5.16, 10.91], P<0.05). Multivariate Logistic regression analysis showed that female ( OR=2.533, 95% CI: 1.306-4.910, P=0.006), atrial fibrillation history ( OR=1.999, 95% CI: 1.044-3.827, P=0.037), neutrophil count within 24 hours of thrombectomy ( OR=1.162, 95% CI: 1.055-1.279, P=0.002), and symptomatic intracranial hemorrhage ( OR=4.066, 95% CI: 1.897-8.718, P<0.001) were independent risk factors for in-hospital death after complete recanalization; risk prediction model, accordingly, was 0.929×female+0.692×atrial fibrillation history+0.150×neutrophil count+1.403×symptomatic intracranial hemorrhage-5.349 ( P: probability of event occurrence). Area under ROC curve of the model was 0.765 (95% CI: 0.689-0.842, P<0.001); calibration curve and Hosmer-Lemeshow test of the model showed good accuracy ( χ2=7.656, P=0.468); decision curve of the model showed good clinical utility at threshold probability of 0.05-0.90. Conclusion:For patients with acute large vessel occlusion stroke at the anterior circulation complicated with atrial fibrillation, symptomatic intracranial hemorrhage or elevated neutrophil count within 24 hours of thrombectomy, or female patients with acute large vessel occlusion stroke at the anterior circulation, in-hospital death still needs to be highly alert after complete recanalization by mechanical thrombectomy.

Objective:To conduct a scope review of perinatal depression symptoms network analysis.Methods:A computer search was conducted on China National Knowledge Infrastructure (CNKI), Wanfang Data, VIP, China Biology Medicine disc, Embase, PubMed, Web of Science, EBSCO, and OVID, with the search period from the establishment of databases to June 1, 2023. Two researchers independently screened the literature and extracted data for the scope review.Results:A total of eight articles were included, of which five were cross-sectional studies and three were longitudinal studies. Regarding central symptoms, two articles identified "worry" as the central symptom, while six articles identified "sadness" as the common central symptom. As for bridging symptoms, "equality with husband" "self-harm" "anxiety" and "discrimination" were considered bridging symptoms. The influencing factors for perinatal depression symptoms included anxiety, family relationships, child emotional difficulties, biomarkers, and discrimination.Conclusions:Nurses can use network analysis methods to gain an in-depth understanding of the perinatal depression symptoms network and implement personalized nursing interventions.

In-fusion cloning technology,as a revolutionary and efficient molecular biology tool,has been applied in multiple research fields such as basic biology,biotechnology,and biomedicine.In this article,we introduce a teaching reform project suitable for undergraduate students in the course of"Molecular Bi-ology Laboratory",which utilizes in-Fusion cloning technology to construct a prokaryotic expression vector for alkaline phosphatase mutant genes.Through specific teaching cases,we systematically explored the design and implementation of experimental projects,and focused on analyzing the key and difficult points of the teaching content.Our teaching practice has found that the implementation of this educational re-form project has achieved very good results in enhancing students' core biological literacy,bioinformatics skills,research thinking,and innovation abilities.At the same time,the application of this technology can significantly improve the quality of experimental teaching,providing new ideas and practical refer-ences for promoting the reform and innovation of National First-Class Courses.

Objective:To construct a training program for nurses in shared-decision making of breast cancer surgery based on core competence, and to provide reference for clinical training.Methods:From February to May 2023, through literature review, qualitative interview, group discussion, Delphi and analytic hierarchy process, the training program were determined.Results:Among 18 experts, there were 3 males and 15 females, aged (44.67±9.24) years old. The final training program included 6 first-level indicators(admission criteria, training content (theory + practice), training format, training hours, training instructors, training evaluation), 17 second-level indicators, and 57 third-level indicators. The effective response rates of 2 rounds of expert consultations were both 18/18, and the expert authority coefficients were both 0.933. The Kendall coordination coefficients of the first and second round were 0.160, 0.213 (both P<0.05). Conclusions:The training program of decision aid nurses in breast cancer surgery is scientific, practical and clinically valuable, and can provide reference for the development of standardized training for nurses.