ObjectiveTo construct a stable monoclonal human embryonic kidney 293 （HEK293） cell line expressing recombinant human coagulation factor Ⅶ （rhFⅦ） and evaluate the expression level and procoagulant bioactivity of rhFⅦ. MethodsThe plasmid pCDNA3.1-EGFP-FⅦ was transfected into HEK293 cells to verify the effectiveness of the transfection system. The plasmid pCDNA3.1-FⅦ was transfected into HEK293 cells， and monoclonal stable transfected cell lines were selected using geneticin （G418）. The transcription of the FⅦ gene was identified by reverse transcription polymerase chain reaction （RT-PCR）. The expression level of rhFⅦ in the supernatant of the monoclonal stable transfected cell line was detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot. The concentration of rhFⅦ was determined by enzyme-linked immunosorbent assay （ELISA）， and the procoagulant activity of rhFⅦ was measured by human coagulation factor Ⅶ potency assay. ResultsHEK293 cells transfected with pcDNA3.1-EGFP-FⅦ showed green fluorescence， indicating that rhFⅦ was successfully expressed in the supernatant of HEK293 cells after transient transfection with pcDNA3.1-FⅦ. The monoclonal stable transfected cell line was obtained by G418 screening. RT-PCR identified that the FⅦ gene was integrated into the genome of the monoclonal stable transfected cell line. The cell viability was good as detected by Cell Counting Kit-8， and a single band of rhFⅦ was obtained by purification of the cell supernatant. The highest rhFⅦ expression was （1.27±0.09） mg/L， and the highest procoagulant activity was （380.29±13.80）%. ConclusionThe monoclonal HEK293 cell lines which can express rhFⅦ protein efficiently and stably with excellent procoagulant bioactivity is successfully screened.

The Pharmacovigilance Guidelines for Clinical Application of Oral Chinese Patent Medicines （hereinafter referred to as the Guidelines） is first specialized in the field of drug safety for oral Chinese patent medicines （OCPMs） in China. Rooted in China's healthcare context， the Guidelines address the unique usage patterns and risk characteristics of OCPMs， filling a regulatory gap in the pharmacovigilance framework specific to this category. To facilitate accurate understanding and effective implementation of the Guidelines， and to promote the standardized development of pharmacovigilance practices for OCPMs， this study offered a systematic interpretation based on its three core components. In the domain of risk monitoring and reporting， the paper analyzed the rationale for multi-source information integration and clarified the criteria for identifying key products and target populations for intensive monitoring. Regarding risk assessment， the Guidelines were examined from three dimensions of formulation components， medication behaviors， and population to address complex safety issues arising from medicinal constituents， irrational use， and individual susceptibility. In the area of risk control， the analysis focused on context-based interventions and dynamic closed-loop management strategies， exploring practical pathways to shift from passive response to proactive risk mitigation. Furthermore， this paper evaluated the applied value of the Guidelines and identified implementation challenges， such as insufficient capacity at the primary-care level and limited digital infrastructure. In response， the study proposed optimization strategies including establishing a dynamic updating mechanism， strengthening training at the grassroots level， and incorporating artificial intelligence to enhance pharmacovigilance capacity. This interpretation aims to provide actionable insights for marketing authorization holders （including manufacturers）， pharmaceutical distributors， healthcare institutions， and research organizations， ultimately supporting the establishment and refinement of a full lifecycle pharmacovigilance system for OCPMs.

With the rapid development of the interdisciplinary area of artificial intelligence and medi-cine,large language model(LLM)has been widely used in the fields such as diagnosis and treatment,medi-cine,and healthcare.LLM has unique advantages in the field of traditional Chinese medicine(TCM),such as high consistency with the"Four Diagnostic Methods",perfect combination of natural language and self-super-vised learning in TCM,the ability to adapt to the characteristics TCM formulas,and the assistance in TCM di-agnosis and treatment.At present,various LLM models have been developed,including the"Qihuang Ask Big Model"and the Digital Traditional Chinese Medicine Big Model"GLM-130B",but they still face challenges such as value mismatch and medical abuse,increased demand for interpretability,lack of advanced technolo-gy,and domestic policy access.This article reviews the evolution of LLM,its unique advantages and applica-tions in the field of TCM,the problems and challenges,and the future development trends,in order to provide reference for the further promotion of LLM in traditional medicine.

Objective To investigate the population density and seasonal fluctuations of Aedes albopictus in Guangzhou City, Guangdong Province, from 2021 to 2023, so as to provide insights into A. albopictus control and management of dengue fever. Methods The surveillance of A. albopictus density was performed in all surveillance sites assigned across all streets (townships) in Guangzhou City during the period from January to December from 2021 to 2023. The surveillance frequency was twice every half month from May to September, and once every month for the rest of a year. In each surveillance period, A. albopictus mosquito larvae were captured from indoor and outdoor small water containers in residential areas, parks, medical facilities, schools, other government sectors and social organizations, construction sites, special industries and others for mosquito species identification. Adult mosquitoes were captured using electric mosquito suction apparatus for species identification and gender classification. Adult mosquitoes and mosquito eggs were collected with mosquito and egg traps at the breeding and dwelling places of Aedes mosquitoes for identification. The mosquito oviposition index (MOI), Breteau index (BI), adult mosquito density index (ADI) and standard space index (SSI) were calculated. The A. albopictus density was classified into grades 0, 1, 2 and 3 in each surveillance site, with Grade 0 density defined eligible, and the eligible rate of A. albopictus density was calculated at all surveillance sites each year from 2021 to 2023. In addition, the changing trends in MOI, SSI, BI and ADI of A. albopictus were analyzed in Guangzhou City from 2021 to 2023. Results The eligible rates of A. albopictus density were 61.69%, 68.75% and 55.15% in surveillance sites of Guangzhou City from 2021 to 2023 (χ² = 297.712, P < 0.001), and appeared a tendency towards a reduction followed by a rise each year, which gradually reduced since January, maintained at a low level during the period between May and October, and gradually increased from November to December. The MOI, SSI, BI and ADI of A. albopictus all appeared a tendency towards a rise followed by a reduction in Guangzhou City during the period between January and December from 2021 to 2023. The BI of A. albopictus peaked in the first half of June in 2021 (4.03), the first half of July in 2022 (3.89) and the last half of August in 2023 (5.02), and the SSI of A. albopictus peaked in the last half of June in 2021 (0.93), the last half of May in 2022 (0.59), and the last half of June (0.94) and the first half of September in 2023 (1.12). In addition, the MOI of A. albopictus peaked in the first half of May in 2021 (8.64), the first half of June in 2022 (8.96), and the last half of May (10.21) and the last half of June in 2023 (10.89), and the ADI of A. albopictus peaked in the first half of June in 2021 (3.41), the last half of June in 2022 (4.06), and the first half of July in 2023 (3.61). Conclusions The density of A. albopictus is high in Guangzhou City during the period from May to October, and the risk of local outbreak caused by imported dengue fever is high. Persistent intensified surveillance of the density and seasonal fluctuation of A. albopictus is recommended and timely mosquito prevention and control is required according to the fluctuation in the A. albopictus density.

Objective:To analyze and compare the reporting data of adverse events following immunization (AEFI) of influenza vaccines in Fujian Province from 2019 to 2023.Methods:Using the National Immunization Program Information Management System, the AEFI reports and vaccination data of influenza vaccines in Fujian Province from 2019 to 2023 were collected, and the reporting rates and clinical characteristics of AEFI of 6 types of influenza vaccines were compared. The 6 types of vaccines in the analysis were as follows: trivalent inactivated influenza vaccines (IIV3) for 6-35 months old people, IIV3 for ≥3 years old people, trivalent live attenuated nasal spray vaccine (LAIV3) for 3-17 years old people, quadrivalent inactivated influenza vaccines (IIV4) for 6-35 months old people, IIV4 for ≥6 months old people, and IIV4 for ≥3 years old people.Results:From 2019 to 2023, a total of 87 687.21 million doses of influenza vaccine were vaccinated in Fujian Province, and 510 cases of AEFI were reported, with a reporting rates of 5.82 per 100 000 doses. Among the 510 cases, 443 (86.86%) were general reactions, 56 (10.98%) were abnormal reactions, 1 (0.20%) was psychogenic reactions, and 10 (1.96%) were coincidence. There were no reports of vaccination accidents and vaccine quality accidents. The reporting rates of AEFI were relatively higher in 2019 and 2020 (18.38 and 18.00 per 100 000 doses, respectively), and lower in 2021, 2022 and 2023 (8.91, 10.68 and 2.30 per 100 000 doses, respectively); the differences were statistically significant (all P<0.05). The differences of reporting rates of AEFI between IIV3 for 6-35 months old people and IIV4 for 6-35 months old people, the injectable vaccines and nasal spray vaccines were not statistically significant. However, the reporting rates of overall AEFI, general reactions and abnormal reactions of IIV3 for ≥3 years old people were all higher than those of IIV4 for ≥3 years old people (7.77 per 100 000 doses vs. 3.88 per 100 000 doses, 6.18 per 100 000 doses vs. 3.59 per 100 000 doses, 1.41 per 100 000 doses vs. 0.19 per 100 000 doses). The reporting rates of overall AEFI and general reaction of IIV3 for 6-35 months old people were both higher than those of IIV3 for ≥3 years old (16.47 per 100 000 doses vs. 7.77 per 100 000 doses, 13.05 per 100 000 doses vs. 6.18 per 100 000 doses), and the differences were statistially significant (all P<0.05). The reporting rates of general abnormal reactions of IIV4 for 6-35 months old and ≥ 6 months old people were both higher than those of IIV4 for ≥3 years old people (14.73 per 100 000 doses and 9.52 per 100 000 doses vs. 3.88 per 100 000 doses); the reporting rates of general reactions and abnormal reactions of IIV4 for ≥6 months old people were both higher than those of IIV4 for ≥3 years old people (12.94 per 100 000 doses vs. 3.59 per 100 000 doses, 1.34 per 100 000 doses vs. 0.19 per 100 000 doses), the differences were statistcially significant (all P<0.05). In terms of clinical features, the reporting rates of fever (37.6-38.5 ℃ and ≥ 38.5 ℃), local redness and swelling (diameter 2.6-5.0 cm), and local induration (diameter ≤2.5 cm and 2.6-5.0 cm) after vaccination of IIV3 for ≥3 years old people were higher than those of IIV4 for ≥ 3 years old people (1.41 per 100 000 doses vs. 0.64 per 100 000 doses, 3.00 per 100 000 doses vs. 1.16 per 100 000 doses); the reporting rates of allergic rash and angioedema of IIV3 for ≥ 3 years old people were higher than those of IIV4 for ≥3 years old people (0.53 per 100 000 doses vs. 0.12 per 100 000 doses, 0.35 per 100 000 doses vs. 0); the differences were statistically significant (all P<0.016 7). Conclusions:The reporting rates of AEFI for influenza vaccines in Fujian Province from 2019 to 2023 was showing a downward trend. The AEFI was mainly general reactions. The reporting rates of AEFI were different among different influenza vaccines, but the overall safety was good.

Objective:To develop an electronic information platform-based assessment tool for entrustable professional activities (EPAs) of master of nursing specialist (MNS) students.Methods:The study used the content analysis method to construct the draft of the assessment tool, which was finalized using the expert meeting method. An electronic information platform was developed based on the assessment tool. The functions of the platform was optimized using the focus group interview method.Results:Based on previous research on the EPA framework for MNS students, an EPA assessment tool was developed, including an assessment form and an assessment protocol. The assessment form consisted of three parts: description of the EPAs, supervision level, and narrative feedback. The assessment protocol covered the assessment objects, assessment initiation strategy, assessment implementation-entrustment decision making, assessment cycle, and quality control. The electronic information platform includes both web-based and mobile app interfaces, with user roles for clinical instructors, graduate students, and project administrators, and with functions such as assessment application, assessment initiation, historical assessment review, and assessment information management.Conclusions:The study developed an EPA assessment tool for MNS students. The tool relies on the convenience and efficiency of the electronic information platform, facilitates clinical instructors in observing the comprehensive performance of MNS students in multiple competencies, and promotes postgraduate students' professional development. Future research will further explore the effectiveness of this tool in assessing clinical practice competencies of MNS students, with the aim of advancing the practice of EPAs in nursing masters' education.

Objective:To evaluate the safety and efficacy of combined inflatable mediastinoscopy with laparoscopy guided by the concept of “reduced field and port” during esophagectomy for esophageal cancer.Methods:This is a retrospective cohort study. The clinical data of 497 patients with esophageal squamous cell carcinoma who underwent minimally invasive esophagectomy at the Center of Minimally Invasive Thoracic Surgery, the Second Affiliated Hospital of Naval Medical University, between January 2017 and December 2024 were retrospectively analyzed. There were 416 male and 81 female patients, with an age of (68.3±8.0) years (range: 44 to 89 years). Patients were divided into the traditional video-assisted thoracoscopic surgery group (Group A, n=354) and the combined inflatable mediastinoscopy with laparoscopic surgery group(Group B, n=143) based on the surgical approach. Furthermore, Group B was subdivided into the multiport laparoscopic group (Group B1, n=81) and the single-incision laparoscopic surgery plus one port group (Group B2, n=62). Perioperative indicators and postoperative survival differences were compared between the groups. Inter-group comparisons were performed using the independent sample t-test, χ2 test, or Fisher′s exact probability test. Survival curves were plotted using the Kaplan-Meier method, and the Log-rank test was used to analyze the survival differences between groups. Results:Compared with Group A, Group B demonstrated a significantly shorter operative time ((181.8±11.4) minutes vs. (196.7±8.1)minutes, t=16.09, P<0.01), a lower incidence of postoperative pulmonary complications (8.4% (12/143) vs. 17.8% (63/354), χ2=6.27, P=0.012), lower perioperative mortality (0 vs. 3.1%(11/354), P=0.039), and a shorter postoperative hospital stay ((16.2±2.2)days vs. (18.9±4.1)days, t=8.56, P<0.01). There was no significant difference in the anastomotic leak rate, number of lymph nodes dissected, or intraoperative blood loss between the two groups (all P>0.05). Overall survival time and recurrence-free survival time showed no significant difference between the two groups (all P>0.05). Subgroup analysis revealed no significant differences in perioperative indicators or postoperative complication rates between Group B1 and Group B2. Conclusions:Compared with traditional thoracoscopic combined with laparoscopic surgery, inflatable mediastinoscopy offered advantages in terms of lower postoperative pulmonary complication rates, shorter operative time, reduced postoperative hospital stay, and lower perioperative mortality. The “reduced field and port” concept could further minimize surgical trauma during the transmediastinal approach for esophagectomy while ensuring surgical safety and efficacy.

Objective:To evaluate the clinical safety of Fufang E'jiao syrup and provide reference for its rational and safe clinical use. Methods:The literature involving Fufang E'jiao syrup in domestic and international databases, as well as the relevant clinical trials on ClinicalTrials.gov and the Chinese Clinical Trial Registry website were searched up to June 1, 2024. Those literature and clinical trials reporting drug adverse events were included, and the basic information about literature/clinical trials (title, publication year, study design, etc.), patients (age, gender, primary diseases, and dosage of Fufang E'jiao syrup), and adverse events (time of occurrence, clinical manifestations, and outcomes) was extracted. The adverse events were standardized and classified using the Medical Dictionary for Regulatory Activities version 25.0, and were also analyzed based on traditional Chinese medicine theory. Results:A total of 19 literature were included in the analysis, including 16 observational/experimental clinical studies, and 3 case reports. The 19 literature reported a total of 430 adverse events involving 398 patients, and the patients were mainly with malignant tumors and anemia. The 430 adverse events involved 11 system organ classes, which mainly included gastrointestinal disorders (260 events, 60.47%, with the most common symptom being dry mouth), respiratory, thoracic, and mediastinal disorders (119 events, 27.67%, with the most common symptom being dry throat), and skin and subcutaneous tissue disorders (16 events, 3.72%, with the most common symptom being mucosal ulcers). Based on traditional Chinese medicine theory, the 430 adverse events were mainly manifested as symptoms of indigestion (nausea, epigastric discomfort, and decreased appetite) and symptoms of "heat" (dry mouth and dry throat).Conclusions:Fufang E'jiao syrup has a relatively good overall safety profile, with the most common adverse events being symptoms of "heat" and gastrointestinal reactions. Patients should not use it blindly, and it should be used with syndrome differentiation in clinical practice.

Objective:To investigate the clinical efficacy and safety of a modified injection technique for improving temporal concavity deformities.Methods:A prospective study was conducted on 157 female patients who underwent temporal concavity augmentation at the Department of Plastic Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, from June 2019 to June 2022. The patients were aged from 32 to 48 (38.2±3.1) years. Granular fat was obtained through liposuction and purification. Using the midpoint of the frontotemporal line as the entry point, the granular fat tissue was injected evenly at multiple points under the skin and superficial temporal fascia. Additional granular fat was injected in the temporal edge area to ensure a smooth appearance. The visual analogue scale (VAS) was used to assess the patients' intraoperative pain levels. Follow-up visits were conducted for one year postoperatively to assess patient satisfaction with the improvement in temporal concavity, local pigmentation, and fine lines in the lateral orbital area based on preoperative and postoperative photographs. Complications, including swelling, ecchymosis, hematoma, fat embolism, infection, fat liquefaction, and subcutaneous induration, were recorded immediately after surgery and at the one-year follow-up.Results:All the 157 patients experienced mild swelling in the injection area postoperatively, which resolved within 3 to 6 days. Ecchymosis occurred in 15 patients and spontaneously resolved within 10 days. Eleven patients had asymmetry in the filled area, with 3 recovering after local massage and observation, and the remaining 8 undergoing a second fat grafting procedure. A total of 25 patients underwent more than two fat grafting procedures, including 4 who underwent three procedures. Mild local hematoma occurred in 3 patients postoperatively. The intraoperative pain VAS was (1.8±0.5) score. After treatment, the improvement scores for temporal concavity, local pigmentation, and fine lines in the lateral orbital area were (7.4±0.7), (6.8±0.7), and (7.9±0.7) scores, respectively. After one year of follow-up, no complications such as fat embolism, infection, fat liquefaction, or subcutaneous induration were observed in the 157 patients.Conclusion:The modified injection technique for improving temporal concavity deformities is safe and effective, with high patient satisfaction.

Gene regulation relies on the precise binding of transcription factors (TFs) at regulatory elements, but simultaneously detecting hundreds of TFs on chromatin is challenging. We developed cFOOT-seq, a cytosine deaminase-based TF footprinting assay, for high-resolution, quantitative genome-wide assessment of TF binding in both open and closed chromatin regions, even with small cell numbers. By utilizing the dsDNA deaminase SsdAtox, cFOOT-seq converts accessible cytosines to uracil while preserving genomic integrity, making it compatible with techniques like ATAC-seq for sensitive and cost-effective detection of TF occupancy at the single-molecule and single-cell level. Our approach enables the delineation of TF footprints, quantification of occupancy, and examination of chromatin influences on TF binding. Notably, cFOOT-seq, combined with FootTrack analysis, enables de novo prediction of TF binding sites and tracking of TF occupancy dynamics. We demonstrate its application in capturing cell type-specific TFs, analyzing TF dynamics during reprogramming, and revealing TF dependencies on chromatin remodelers. Overall, cFOOT-seq represents a robust approach for investigating the genome-wide dynamics of TF occupancy and elucidating the cis-regulatory architecture underlying gene regulation.
Transcription Factors/genetics* ; Humans ; Chromatin/genetics* ; Animals ; Binding Sites ; Mice ; DNA Footprinting/methods*