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

Objective To investigate the effect of pneumoperitoneum pressure during robotic-assisted kidney transplantation (RAKT) on the function of the transplant kidney. Methods The data of 243 kidney transplant recipients were retrospectively analyzed and divided into open kidney transplantation (OKT) group (n=105) and RAKT group (n=138). The RAKT group was further divided into 13 mmHg group (n=67) and 7 mmHg group (n=71) based on pneumoperitoneum pressure. The donor information, recipient's preoperative general data, intraoperative data, and postoperative recovery of the three groups were compared. In the RAKT group, the renal artery, segmental artery, interlobar artery, and venous flow velocity of the transplant kidney were measured using laparoscopic ultrasound. Results There was a statistically significant difference in donor types among the groups (P<0.05), while other donor information and recipient's preoperative general data showed no statistically significant differences (all P>0.05). There were no statistically significant differences in serum creatinine and complications at 30 days and 1 year postoperatively among the groups (all P>0.05). The OKT group and 7 mmHg group had more intraoperative urine output than the 13 mmHg group. Both RAKT groups had less intraoperative blood loss and shorter hospital stays than the OKT group, and longer operation times than the OKT group (all P<0.05). There were no statistically significant differences in operation time, intraoperative blood loss, and hospital stay between the two RAKT groups (all P>0.05). The vascular flow velocity of the transplant kidney decreased at 13 mmHg compared to 7 mmHg pneumoperitoneum pressure, but the differences were not statistically significant (all P>0.05). Conclusions Controllable pneumoperitoneum pressure has a limited impact on the vascular flow velocity of the transplanted kidney. RAKT is a safe and effective surgical method under appropriate pneumoperitoneum pressure, and choosing a lower pneumoperitoneum pressure is more conducive to the early recovery of renal function postoperatively.

The traditional Chinese medicine （TCM） myocardial infarction （MI） unit is a standardized， regulated， and continuous integrated care unit guided by TCM theory and built upon existing chest pain centers or emergency care units. This unit emphasizes multidisciplinary collaboration and forms a restructured clinical entity without altering current departmental settings， offering comprehensive diagnostic and therapeutic services with full participation of TCM in the treatment of MI. Its core medical model is patient-centered and disease-focused， providing horizontally integrated TCM-based care across multiple specialties and vertically constructing a full-cycle treatment unit for MI， delivering prevention， treatment， and rehabilitation during the acute， stable， and recovery phases. Additionally， the unit establishes a TCM-featured education and prevention mechanism for MI to guide patients in proactive health management， reduce the incidence of myocardial infarction， and improve quality of life.

Objective:The aim of this study was to analyze the registration status of osteoporosis drug clinical trials in China, providing a reference for the research and development of therapeutic drugs for osteoporosis.Methods:Using the Drug Clinical Trial Registration and Information Publication Platform on the website of the State Drug Administration as the search platform, we retrieved the information on the public announcement of drug clinical trials with osteoporosis as an indication, and the search period was limited from 2013 to February 2025.Researchers extracted the key information and performed statistics.Results:A total of 182 registered osteoporosis drug clinical trial projects were collected.The main target indications were predominantly osteoporosis in postmenopausal women, and there were relatively few trials targeting the elderly subjects.56.0%(102/182)of the registered trials were bioequivalence studies, and 21.4%(39/182)were phase I clinical studies.Conclusions:The number of registered trials and the research and development process of domestic osteoporosis drug clinical trials in China have shown a stable trend in general, and the types of trials are mainly concentrated in bioequivalence studies and early clinical studies.In order to further meet the medication needs of domestic patients, China still needs to increase the research and development efforts of innovative osteoporosis drugs.

This consensus was developed by the Orthodontic Society of the Chinese Stomatological Association to provide a systematic, scientific, and practical guideline for informed consent in orthodontic care. Orthodontic treatment is typically lengthy, highly individualized, and involves multiple factors such as growth and development, occlusal function, and facial esthetics. Rapid technological advances and diverse risk profiles make the traditional reliance on orthodontist experience or institutional templates insufficient to ensure patients′ full understanding and autonomous decision-making. To address this, the expert panel conducted extensive reviews of domestic and international guidelines, analyzed representative dispute cases, and performed multicenter patient-clinician surveys. Using a multi-round Delphi method, the group established a standardized informed consent framework covering the initial consultation, treatment, and retention phases. The consensus emphasizes that informed consent is not only a fundamental legal and ethical requirement but also a key step in building trust, improving patient compliance, and enhancing treatment satisfaction. Orthodontists should clearly and comprehensively explain treatment plans, potential risks, uncertainties, and associated costs, while respecting the autonomy of patients or guardians, and maintain continuous communication and dynamic evaluation throughout the treatment process. The release of this consensus provides unified and authoritative guidance for clinical orthodontics, helping to standardize informed consent, enhance its transparency, safeguard patient rights, reduce medical risks, and promote high-quality, sustainable development of orthodontic practice.

Objective To explore the predictive value of growth differentiation factor 15(GDF-15)for atrial fibrillation(AF)in patients with coronary heart disease(CHD).Methods A prospective observation cohort of 1261 elderly CHD patients was randomly sampled from the First Medical Center of Chinese PLA General Hospital from January 2012 to December 2015.ELISA was used to detect GDF-15 level in all the subjects,and their baseline data were collected.Until March 2023,93 patients were lost during the follow-up period,and finally 1168 patients completed the follow-up,with a median time of 9.4 years.According to AF occurred or not during the period,the eligible patients were divided into an AF group(197 cases)and a non-AF group(971 cases),and based on their medical history,also assigned into a stable angina pectoris(SAP,n=304)and an acute coronary syndrome group(ACS,n=864).Logistic regression analysis was used to determine whether GDF-15 is a risk factor for AF in elderly CHD patients.ROC curve was plotted to assess the predictive value of GDF-15,guideline recommended CHARGE-AF scoring model,and their combination for AF events in the patients.Results The AF group had significantly advanced age,larger proportions of smoking history,type 2 diabetes,hypertension,old myocardial infarction,stroke,ACS and administration of statins,higher ratio of angiotensin converting enzyme inhibi-tor/angiotensin receptor blocker,and elevated levels of GDF-15 and N-terminal pro-B-type natri-uretic peptide,and lower left ventricular ejection fraction,SAP incidence,and low-density lipopro-tein cholesterol,total cholesterol and triglycerides levels when compared with the non-AF group(P＜0.05,P＜0.01).Multivariate logistic regression analysis showed that GDF-15 was not a risk factor for AF in elderly CHD patients(OR=0.92,95％CI:0.74-1.16,P=0.489),but was a risk factor for AF in SAP patients(OR=1.38,95％CI:1.07-2.79,P=0.015),and in ACS patients,still not a risk factor for AF(OR=0.81,95％CI:0.63-1.05,P=0.814).ROC curve analysis showed that the AUC value of GDF-15 combined with the CHARGE-AF scoring model in predic-ting AF was 0.682 in the elderly CHD patients,0.746 in the SAP patients,and 0.680 in the ACS patients.Conclusion Elevated GDF-15 level is an independent risk factor and predictor of AF in elderly SAP patients.In SAP patients,the combination of GDF-15 and CHARGE-AF scoring model further improves the predictive performance of AF occurrence.

Objective:To summarize the early clinical outcomes of aortic valve replacement(AVR) using the Perceval sutureless aortic valve.Methods:This retrospective study included 50 patients who underwent AVR with the Perceval sutureless prostheses at Beijing Anzhen Hospital between June 2023 and January 2025. Surgical parameters, early clinical outcomes, valve function, and hemodynamic performance were evaluated to summarize clinical effectiveness.Results:The mean age of patients was(63.5±9.1) years, predominantly female(36/50). Severe aortic stenosis was present in 43 cases(86.0%). A preoperative aortic annulus dimension of 20.0(19.0, 21.0) mm measured in both anteroposterior and transverse diameters. Preoperative peak transvalvular gradient was(92.7±31.0)mmHg(1 mmHg=0.133 kPa), with a mean gradient of (58.0±21.2) mmHg. All procedures were successfully completed using the Perceval sutureless prostheses. Isolated AVR was performed in 20 patients(40.0%), with cardiopulmonary bypass and aortic cross-clamp times of 75.0(50.5, 99.5) min and 50.5(29.5, 71.5) min, respectively. Postoperative transesophageal echocardiography revealed an immediate reduction in the peak transvalvular gradient to 11.0(8.0, 18.0) mmHg, significantly lower compared to preoperative measurements( P<0.001). Two cases of paravalvular leakage and one case requiring permanent pacemaker implantation were reported postoperatively. All patients completed the 3-month follow-up, with one death during the follow-up period; the remaining patients exhibited normal prosthetic valve function without major adverse cardiovascular events. Significant postoperative reductions were observed in left ventricular end-diastolic diameter(45.8 mm vs. 43.2 mm, P=0.003) and left atrial diameter(53.9 mm vs. 44.6 mm, P<0.001) compared with baseline. Conclusion:AVR using the Perceval sutureless prostheses demonstrated safe and effective early clinical outcomes with excellent hemodynamic performance and low incidence of postoperative paravalvular leakage and permanent pacemaker implantation. The sutureless technique represents a viable alternative strategy, particularly advantageous for patients with small aortic annuli or complex surgical conditions, warranting broader clinical adoption.

This study aims to investigate the relationship between serum vitamin A(VA)and mar-bling grade and the effect of different levels of serum VA on slaughter performance and fatty acid composition and related gene expression in the longissimus dorsi muscle of Woking black cattle and Angus cattle.Thirty Woking black cattle and seventeen Angus cattle aged 30 months were ran-domly selected and analyzed for the linear relationship between serum VA and marbling grade af-ter slaughter.The cattle were divided into three groups:the low VA group,medium VA group and high VA group,ranked in order of VA value in both Woking black cattle and Angus cattle.Statisti-cal analysis of the effects of different types and levels of VA on marbling grade,slaughter performance,fatty acid composition,and the effects of different levels of VA on the expression of genes related to intramuscular fat deposition and other genes in Woking black cattle or Angus cat-tle were also analyzed.The results showed that the marbling grade of Woking black cattle increased numerically with increasing serum VA at slaughter(P=0.203),whereas Angus cattle showed a numerical decrease(P=0.139).Analyses of subsequent subgroups showed that Woking black cat-tle had significantly higher marbling grade and oleic acid,monounsaturated/saturated fatty acids ratio in the longissimus dorsi muscle compared to Angus cattle(P＜0.05).As serum VA levels in-creased,DHA was significantly higher and n-6/n-3 fatty acid ratio significantly lower in the longis-simus dorsi muscle of Woking black cattle(P＜0.05).Whereas VA was elevated in Angus cattle,a significant decrease in DHA and a significant increase in n-6/n-3 fatty acids(P＜0.05)were found.Furthermore,a notable up-or down-regulation(P＜0.05)of LPL,FABP4,PPARγ,C APZA2 and Villin 2 was observed in Woking black or Angus cattle,respectively,as VA levels increased.Based on these results,it was suggested that Woking black cattle require an appropriate increase in dieta-ry VA during the late fattening stage,which was found to produce a higher marbling grade and a higher percentage of beneficial fatty acids for human health when serum VA reached 80.7 IU/dL.Whereas Angus cattle still need to be restricted in ration VA content in the late fattening stage,when serum VA is elevated to 73.6 IU/dL,they produce beef that not only has a lower marbling grade but also has a corresponding reduction in fatty acids beneficial to human health.

Objective To develop and validate a risk prediction model for infiltration/extravasation in peripheral intravenous catheter therapy.Methods This retrospective study analyzed 942 patients who completed the Infiltration/Extravasation Risk Assessment Form between January and June 2023 at the First Affiliated Hospital of Army Medical University(including Departments of Neurology,Endocrinology,Gastroenterology and Hepatobiliary Surgery).Patients were allocated to a derivation cohort(n=628)and validation cohort(n=314)in a 2∶1 ratio based on catheterization chronology.The derivation cohort served for model development and internal validation,while the validation cohort underwent external validation.Logistic scoring method constructed the risk model,with Hosmer-Lemeshow(HL)test assessing goodness-of-fit and ROC curve evaluating predictive performance.Twenty-one potential risk factors were assessed,including age,gender,chronic diseases,clinician experience,treatment compliance,and total infusion volume.Results Infiltration/extravasation occurred in 48 cases(5.10%incidence:31 derivation/17 validation).Among 21 factors,15 showed significant association(P<0.05),with 6 independent predictors:junior high school education or below(OR=5.2),chronic disease history(OR=3.1),poor compliance(OR=2.8),lower extremity venipuncture(OR=4.1),total infusion≥1 000 mL(OR=3.5),and hyperosmotic/corrosive medications(OR=6.7).The final prediction model was:Y=2×(low education)+1×(chronic disease)+1×(poor compliance)+1×(lower extremity puncture)+1×(volume≥1 000 mL)+2×(corrosive agents).For the derivation cohort,AUC was 0.967(95%CI:0.936～0.998),specificity 0.911,sensitivity 0.935,with good calibration(χ2=4.135,P=0.845).Validation cohort showed AUC 0.939(0.853～1.000),specificity 0.919,sensitivity 0.941,and acceptable calibration(χ2=8.998,P=0.085).Conclusion This model demonstrates excellent discriminative ability and calibration,providing an effective tool for identifying high-risk patients and guiding targeted preventive strategies.

Objective To explore the effects of triptolide(TPL)on the proliferation,migration and invasion of gastric cancer cells and its mechanism.Methods Human gastric cancer cell line MKN45 was cultured in vitro and treated with different concentrations of TPL for 48 hours.The cell proliferation inhibition rate was detected by CCK-8 method and the optimal concentration was selected for subsequent experiments.qRT-PCR was used to detect the expression of miR-29b and KDM2A mRNA in cells treated with different concentrations of TPL.MKN45 cells at logarithmic growth phase were randomly divided into the control group(without any treatment),the TPL group(treated with 200 μg/mL TPL),the inhibitor-NC+TPL group(transfected with inhibitor-NC and then treated with 200 μg/mL TPL),and the miR-29b inhibitor+TPL group(transfected with miR-29b inhibitor and then treated with 200 μg/mL TPL).qRT-PCR was used to detect the expression of miR-29b and KDM2A mRNA in each group of cells,and Western blot was used to detect the expression of KDM2A protein.The clone formation ability of each group of cells was detected by plate clone formation assay,and the migration and invasion abilities of each group of cells were detected by Transwell assay.Results TPL at concentrations of 25 μg/mL,50 μg/mL,100 μg/mL,and 200 μg/mL could significantly inhibit the proliferation of MKN45 cells(P<0.05),up-regulate the expression of miR-29b in cells(P<0.05),and down-regulate the expression of KDM2A mRNA(P<0.05).The effect was most obvious at the concentration of 200 μg/mL,so 200 μg/mL TPL was selected for the subsequent experiments.Compared with the control group,the expression of miR-29b in the TPL group increased(P<0.05),the expression of KDM2A mRNA and protein decreased(P<0.05),and the numbers of clone formation,migration and invasion cells reduced(P<0.05).Compared with the inhibitor-NC+TPL group,the expression of miR-29b in the miR-29b inhibitor+TPL group decreased(P<0.05),the expression of KDM2A mRNA and protein increased(P<0.05),and the numbers of clone formation,migration and invasion cells increased(P<0.05).Conclusion TPL can inhibit the proliferation,migration and invasion of gastric cancer cells,and its mechanism is related to the regulation of the miR-29b/KDM2A signaling pathway.