Constructing an efficient and easy-to-operate multigene expression system is currently a crucial part of plant genetic engineering. In this study, a fragment carrying three independent gene expression cassettes and the expression unit of the gene-silencing suppressor protein(RNA silencing suppressor 19 kDa protein, P19) simultaneously was designed and constructed. This fragment was cloned into the commonly used plant expression vector pCAMBIA300, and the plasmid pC1300-TP2-P19 was obtained. Each gene expression cassette consists of different promoters, fusion tags, and terminators. The target gene can be flexibly inserted into the corresponding site through enzymatic digestion and ligation or recombination and fused with different protein tags, which provides great convenience for subsequent detection. The enhanced green fluorescent protein(eGFP) reporter gene was individually constructed into each expression cassette to verify the feasibility of this vector system. The results of tobacco transient expression and laser-confocal microscopy showed that each expression cassette presented independent and normal expression. Meanwhile, the three key enzyme genes in the betanin synthesis pathway, BvCYP76AD, BvDODA1, and DbDOPA5GT, were constructed into the three expression cassettes. The results of tobacco transient expression phenotype, protein immunoblotting(Western blot), and chemical detection of product demonstrated that the three exogenous genes were highly expressed, and the target compound betanin was successfully produced. The above results indicated that the constructed multigene expression system for plants in this study was efficient and reliable and can achieve the co-transformation of multiple plant genes. It can provide a reliable vector platform for the analysis of plant natural product synthesis pathways, functional verification, and plant metabolic engineering.
Nicotiana/metabolism* ; Genetic Vectors/metabolism* ; Gene Expression Regulation, Plant ; Plant Proteins/metabolism* ; Plants, Genetically Modified/metabolism* ; Genetic Engineering/methods* ; Green Fluorescent Proteins/metabolism* ; Gene Expression

Debates persist regarding the efficacy and safety of azvudine, particularly its real-world outcomes. This study involved patients aged ≥60 years who were admitted to 25 hospitals in mainland China with confirmed SARS-CoV-2 infection between December 1, 2022, and February 28, 2023. Efficacy outcomes were all-cause mortality during hospitalization, the proportion of patients discharged with recovery, time to nucleic acid-negative conversion (T _NANC), time to symptom improvement (T _SI), and time of hospital stay (T _HS). Safety was also assessed. Among the 5884 participants identified, 1999 received azvudine, and 1999 matched controls were included after exclusion and propensity score matching. Azvudine recipients exhibited lower all-cause mortality compared with controls in the overall population (13.3% vs. 17.1%, RR, 0.78; 95% CI, 0.67-0.90; P = 0.001) and in the severe subgroup (25.7% vs. 33.7%; RR, 0.76; 95% CI, 0.66-0.88; P < 0.001). A higher proportion of patients discharged with recovery, and a shorter T _NANC were associated with azvudine recipients, especially in the severe subgroup. The incidence of adverse events in azvudine recipients was comparable to that in the control group (2.3% vs. 1.7%, P = 0.170). In conclusion, azvudine showed efficacy and safety in older patients hospitalized with COVID-19 during the SARS-CoV-2 omicron wave in China.

Objective:To evaluate the application potential of the bimodal ultrasound/near-infrared (NIR) composite nanoscale probe Arg-Gly-Asp (RGD)/Ag 2Te/perfluoropentane (PFP) @ mesoporous silica nanoparticles (MSN) in the diagnosis and treatment of microvascular diseases. Methods:Nanoprobes loaded with RGD, PFP and Ag 2Te were prepared by ultrasound sonication and carbodiimide method. The characterization of the nanoprobes was determined. The imaging performance, photothermal response, and target-seeking ability of the nanoprobes under NIR irradiation were verified. The biosafety of the nanoprobes was examined, and the thrombolytic ability of the nanoprobes was evaluated. The mice were observed to visualize microvessels of the abdominal wall under the NIR-Ⅱ imaging, and the microvascular visualization ability of the nanoprobes was evaluated. Results:The particle size of nanoprobes was (205.3±2.9) nm and the potential was (2.05±0.58) mV. The coupling rate of the RGD was (82.27±0.36)%, the encapsulation rate of the quantum dots was (80.80±3.26)%, and the photostability of the quantum dots was good. The fluorescence intensity was enhanced with the increase of the mass concentration of RGD/Ag 2Te/PFP@MSN, and the warming effect was more obvious. After ultrasound and NIR irradiation, the thrombolysis rate was significantly increased. RGD/Ag 2Te/PFP@MSN successfully realized NIR-Ⅱ fluorescence imaging of mice microvessels. The cytotoxicity assay and hemolysis assay showed that the probe had a good biosafety. Conclusion:The RGD/Ag 2Te/PFP@MSN nanoprobe is a potential strategy for targeted therapy of thrombotic diseases, combining dual-modality therapy of ultrasound and NIR to offer new possibilities for non-invasive and visual diagnosis and treatment of microvascular embolism.

Objective: To explore the association between dietary patterns and overweight and obesity among primary and middle school students, so as to provide the guidance of diet balance and obesity prevention for students. Methods: Students from 11 primary and middle schools in Tongzhou District, Beijing Municipality, were selected using the stratified cluster sampling method. Demographic information was collected through a general questionnaire, and dietary intake types and frequencies over the past week were assessed using a food frequency questionnaire. Dietary patterns were determined using factor analysis, and the dietary pattern factor scores were divided into Q1, Q2, Q3, and Q4 groups based on quartiles. Height and weight were collected through physical examinations, and Z-scores of body mass index adjusted for gender and age were calculated to assess overweight and obesity. The association between dietary patterns and overweight and obesity was analyzed using a multivariable logistic regression model. Results: A total of 1 485 students were surveyed, including 745 males (50.17%) and 740 females (49.83%). The mean age was (12.87±2.86) years. Three dietary patterns were identified: animal and plant protein dietary pattern, high-fat and high-sugar dietary pattern, and vegetable-fruit-dairy dietary pattern. The detection rates of overweight and obesity were 42.57%, 48.42%, and 34.75%, respectively, with statistically significant differences (P<0.05). Multivariable logistic regression analysis showed that compared with the vegetable-fruit-dairy dietary pattern, the animal and plant protein dietary pattern (OR=1.406, 95%CI: 1.084-1.823) and the high-fat and high-sugar dietary pattern (OR=2.137, 95%CI: 1.643-2.779) were associated with a higher risk of overweight and obesity among primary and middle school students. Compared with the Q1 group of dietary pattern factor scores, the Q3 group (OR=1.631, 95%CI: 1.206-2.208) and Q4 group (OR=1.965, 95%CI: 1.446-2.671) of the high-fat and high-sugar dietary pattern had an increased risk of overweight and obesity, while the Q4 group of the vegetable-fruit-dairy dietary pattern (OR=0.551, 95%CI: 0.406-0.747) had a reduced risk of overweight and obesity. There was no statistical association between factor scores of animal and plant protein dietary pattern and overweight and obesity (all P>0.05). Conclusions The animal and plant protein dietary pattern and the high-fat and high-sugar dietary pattern are associated with a higher risk of overweight and obesity among primary and middle school students. Within the same dietary pattern, the more inclined students are to the high-fat and high-sugar dietary pattern, the higher their risk of overweight and obesity, while the more inclined they are to the vegetable-fruit-dairy dietary pattern, the lower their risk of overweight and obesity.

Objective:To investigate the long-term efficacy and influencing factors of transcatheter adrenal ablation in patients with primary aldosteronism (PA).Methods:This cohort study retrospectively enrolled PA patients who underwent transcatheter adrenal ablation at Daping Hospital, Army Medical University between January 2021 and December 2024. According to PASO criteria, patients were categorized into groups based on clinical outcomes (complete, partial, or no remission), biochemical outcomes (complete, partial, or no remission), and composite outcomes (complete or incomplete remission). All participants underwent 1-year follow-up, with intergroup comparisons of clinical characteristics and surgical approaches. Multivariate logistic regression models were used to identify factors influencing long-term efficacy post-transcatheter adrenal ablation in PA patients.Results:A total of 122 PA patients were enrolled, aged (47.7±11.1) years, including 55 males (45.1%). Baseline aldosterone-to-renin ratio was 0.43(0.19,0.86)(pmol·L -1)/(μU·L -1). Bilateral adrenal lesions were present in 33 cases (27.1%), while 70 (57.4%) had nodules or adenomas. Adrenal venous sampling confirmed lateralized hypersecretion in 107 patients (87.7%, left or right dominance). According to PASO criteria, 93.4% (114/122) and 95.1% (116/122) of patients achieved complete or partial remission in biochemical and clinical parameters at 1-year post-ablation, respectively. For biochemical outcomes: 40 complete, 74 partial, and 8 no remission. Patients in the partial-remission group were older than those in the no-remission group ((49.4±11.2) vs. (39.6±9.8) years), while complete-remission group had higher bilateral non-lateralized secretion rates than partial remission group (27.5% vs. 4.1%, both P<0.05). For clinical outcomes: 26 complete, 90 partial, 6 no remission. Compared to complete-remission group, partial-remission group had higher male proportion (51.1% vs. 26.9%), longer hypertension duration (4.0 (0.7, 10.0) years vs. 1.5 (0.1, 5.0) years), but lower office diastolic blood pressure ((88±11) mmHg vs. (94±12 mmHg), 1 mmHg=0.133 kPa, all P<0.05). For composite outcomes: 56 complete and 66 incomplete remission. Compared with incomplete remission group, complete remission group had lower prevalence of diabetes (8.9% vs. 22.7%) and higher proportion of bilateral non-lateralized secretion (21.4% vs. 4.6%, both P<0.05). Multivariate logistic regression identified diabetes ( OR=3.635, 95% CI 1.029-12.834, P=0.045) and lateralized secretion ( OR=9.056, 95% CI 2.039-40.212, P=0.004) as independent risk factors for poor composite outcomes after transcatheter adrenal ablation in PA patients, whereas higher office diastolic blood pressure acts as a protective factor ( OR=0.957, 95% CI 0.925-0.992, P=0.015). Conclusion:One year after transcatheter adrenal ablation, the majority of patients achieved complete or partial remission in biochemical and clinical parameters.Patients with non-lateralized adrenal hypersecretion demonstrated a higher likelihood of sustained biochemical remission and superior composite outcomes compared to those with lateralized hypersecretion.

Spine fracture and dislocation are common traumatic spinal conditions that often require surgical intervention due to compromised spinal stability. Surgical approaches include anterior, posterior, and combined anterior-posterior spinal procedures. According to the specific surgical requirements, patients may be placed in the prone position or repositioned between prone and supine positions during surgery. Intraoperative repositioning has become an essential step in patient positioning. However, during repositioning, patients with spinal fracture and dislocation are at increased risk for complications such as hemodynamic instability, nerve injury, and pressure injuries to the skin and soft tissue. Notably, due to the instability of the spinal cord, even minor manipulations can further exacerbate the damage, potentially leading to severe outcomes like paraplegia. Although the current clinical guidelines provide instructive recommendations for standard position, there remains no specific protocols for intraoperative repositioning in patients with spine fracture and dislocation. With a concern for the lack of clinical studies on positioning techniques, risk prevention, and operational norms for special patients, no applicable guidelines or standards are available. A consensus was required to provide clinical reference, meet the requirements of surgical treatment, and minimize the safety risks of patients caused by improper placement of positions. Professional Committee of Operating Room Nursing of Shaanxi Nursing Association organized experts in nursing management and operating room nursing from major hospitals across China to formulate Expert consensus on intraoperative repositioning for patients with spinal fracture and dislocation ( version 2025). The consensus provides 11 recommendations covering pre-repositioning preparation, intraoperative maneuvers, and post-repositioning observation, aiming to provide references for clinical standardization of the intraoperative repositioning process and protection of patients′ safety.

Spine fracture and dislocation are common traumatic spinal conditions that often require surgical intervention due to compromised spinal stability. Surgical approaches include anterior, posterior, and combined anterior-posterior spinal procedures. According to the specific surgical requirements, patients may be placed in the prone position or repositioned between prone and supine positions during surgery. Intraoperative repositioning has become an essential step in patient positioning. However, during repositioning, patients with spinal fracture and dislocation are at increased risk for complications such as hemodynamic instability, nerve injury, and pressure injuries to the skin and soft tissue. Notably, due to the instability of the spinal cord, even minor manipulations can further exacerbate the damage, potentially leading to severe outcomes like paraplegia. Although the current clinical guidelines provide instructive recommendations for standard position, there remains no specific protocols for intraoperative repositioning in patients with spine fracture and dislocation. With a concern for the lack of clinical studies on positioning techniques, risk prevention, and operational norms for special patients, no applicable guidelines or standards are available. A consensus was required to provide clinical reference, meet the requirements of surgical treatment, and minimize the safety risks of patients caused by improper placement of positions. Professional Committee of Operating Room Nursing of Shaanxi Nursing Association organized experts in nursing management and operating room nursing from major hospitals across China to formulate Expert consensus on intraoperative repositioning for patients with spinal fracture and dislocation ( version 2025). The consensus provides 11 recommendations covering pre-repositioning preparation, intraoperative maneuvers, and post-repositioning observation, aiming to provide references for clinical standardization of the intraoperative repositioning process and protection of patients′ safety.

Objective:To evaluate the application potential of the bimodal ultrasound/near-infrared (NIR) composite nanoscale probe Arg-Gly-Asp (RGD)/Ag 2Te/perfluoropentane (PFP) @ mesoporous silica nanoparticles (MSN) in the diagnosis and treatment of microvascular diseases. Methods:Nanoprobes loaded with RGD, PFP and Ag 2Te were prepared by ultrasound sonication and carbodiimide method. The characterization of the nanoprobes was determined. The imaging performance, photothermal response, and target-seeking ability of the nanoprobes under NIR irradiation were verified. The biosafety of the nanoprobes was examined, and the thrombolytic ability of the nanoprobes was evaluated. The mice were observed to visualize microvessels of the abdominal wall under the NIR-Ⅱ imaging, and the microvascular visualization ability of the nanoprobes was evaluated. Results:The particle size of nanoprobes was (205.3±2.9) nm and the potential was (2.05±0.58) mV. The coupling rate of the RGD was (82.27±0.36)%, the encapsulation rate of the quantum dots was (80.80±3.26)%, and the photostability of the quantum dots was good. The fluorescence intensity was enhanced with the increase of the mass concentration of RGD/Ag 2Te/PFP@MSN, and the warming effect was more obvious. After ultrasound and NIR irradiation, the thrombolysis rate was significantly increased. RGD/Ag 2Te/PFP@MSN successfully realized NIR-Ⅱ fluorescence imaging of mice microvessels. The cytotoxicity assay and hemolysis assay showed that the probe had a good biosafety. Conclusion:The RGD/Ag 2Te/PFP@MSN nanoprobe is a potential strategy for targeted therapy of thrombotic diseases, combining dual-modality therapy of ultrasound and NIR to offer new possibilities for non-invasive and visual diagnosis and treatment of microvascular embolism.

Objective:To investigate the long-term efficacy and influencing factors of transcatheter adrenal ablation in patients with primary aldosteronism (PA).Methods:This cohort study retrospectively enrolled PA patients who underwent transcatheter adrenal ablation at Daping Hospital, Army Medical University between January 2021 and December 2024. According to PASO criteria, patients were categorized into groups based on clinical outcomes (complete, partial, or no remission), biochemical outcomes (complete, partial, or no remission), and composite outcomes (complete or incomplete remission). All participants underwent 1-year follow-up, with intergroup comparisons of clinical characteristics and surgical approaches. Multivariate logistic regression models were used to identify factors influencing long-term efficacy post-transcatheter adrenal ablation in PA patients.Results:A total of 122 PA patients were enrolled, aged (47.7±11.1) years, including 55 males (45.1%). Baseline aldosterone-to-renin ratio was 0.43(0.19,0.86)(pmol·L -1)/(μU·L -1). Bilateral adrenal lesions were present in 33 cases (27.1%), while 70 (57.4%) had nodules or adenomas. Adrenal venous sampling confirmed lateralized hypersecretion in 107 patients (87.7%, left or right dominance). According to PASO criteria, 93.4% (114/122) and 95.1% (116/122) of patients achieved complete or partial remission in biochemical and clinical parameters at 1-year post-ablation, respectively. For biochemical outcomes: 40 complete, 74 partial, and 8 no remission. Patients in the partial-remission group were older than those in the no-remission group ((49.4±11.2) vs. (39.6±9.8) years), while complete-remission group had higher bilateral non-lateralized secretion rates than partial remission group (27.5% vs. 4.1%, both P<0.05). For clinical outcomes: 26 complete, 90 partial, 6 no remission. Compared to complete-remission group, partial-remission group had higher male proportion (51.1% vs. 26.9%), longer hypertension duration (4.0 (0.7, 10.0) years vs. 1.5 (0.1, 5.0) years), but lower office diastolic blood pressure ((88±11) mmHg vs. (94±12 mmHg), 1 mmHg=0.133 kPa, all P<0.05). For composite outcomes: 56 complete and 66 incomplete remission. Compared with incomplete remission group, complete remission group had lower prevalence of diabetes (8.9% vs. 22.7%) and higher proportion of bilateral non-lateralized secretion (21.4% vs. 4.6%, both P<0.05). Multivariate logistic regression identified diabetes ( OR=3.635, 95% CI 1.029-12.834, P=0.045) and lateralized secretion ( OR=9.056, 95% CI 2.039-40.212, P=0.004) as independent risk factors for poor composite outcomes after transcatheter adrenal ablation in PA patients, whereas higher office diastolic blood pressure acts as a protective factor ( OR=0.957, 95% CI 0.925-0.992, P=0.015). Conclusion:One year after transcatheter adrenal ablation, the majority of patients achieved complete or partial remission in biochemical and clinical parameters.Patients with non-lateralized adrenal hypersecretion demonstrated a higher likelihood of sustained biochemical remission and superior composite outcomes compared to those with lateralized hypersecretion.

Knowledge about the neuronal dynamics and the projectome are both essential for understanding how the neuronal network functions in concert. However, it remains challenging to obtain the neural activity and the brain-wide projectome for the same neurons, especially for neurons in subcortical brain regions. Here, by combining in vivo microscopy and high-definition fluorescence micro-optical sectioning tomography, we have developed strategies for mapping the brain-wide projectome of functionally relevant neurons in the somatosensory cortex, the dorsal hippocampus, and the substantia nigra pars compacta. More importantly, we also developed a strategy to achieve acquiring the neural dynamic and brain-wide projectome of the molecularly defined neuronal subtype. The strategies developed in this study solved the essential problem of linking brain-wide projectome to neuronal dynamics for neurons in subcortical structures and provided valuable approaches for understanding how the brain is functionally organized via intricate connectivity patterns.
Animals ; Neurons/physiology* ; Mice ; Brain/physiology* ; Mice, Inbred C57BL ; Somatosensory Cortex/physiology* ; Neural Pathways/physiology* ; Hippocampus/physiology* ; Mice, Transgenic ; Male ; Brain Mapping ; Nerve Net/physiology* ; Substantia Nigra/physiology* ; Tomography, Optical/methods*