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.

ObjectiveTo investigate whether Bushen Huoxue prescription improves embryo implantation through regulating exosomal miRNA to enhance maternal-fetal interface communication based on Bushen Huoxue therapy. MethodsIn the animal experiment， all the rats （except for the blank group） were administered hydroxyurea （450 mg·kg^-1） via gavage for 10 d， as well as epinephrine （0.3 mg·kg^-1） and mifepristone （5.5 mg·kg^-1） via subcutaneous injection for 7 d to establish an implantation disorder model of kidney deficiency and blood stasis type. The Bushen Huoxue prescription （BSHX） groups were administered the prescription at different doses （7.30 g·kg^-1 for the high-dose group， 3.65 g·kg^-1 for the medium-dose group， and 1.83 g·kg^-1 for the low-dose group） via gavage. The dydrogesterone group was administered the corresponding medicine （2.63 mg·kg^-1） via gavage. After intervention for 10 days， uterine histopathological changes were observed via hematoxylin-eosin （HE） staining. Mucin （MUC1）， forkhead box protein O1 （FoxO1）， and homeobox A10 （HoxA10） expression levels were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. Cell experiment selected primary endometrial epithelial cells （EEC） and trophoblast cells （TC） as research subjects. Exosome-free medicated serum was prepared by ultracentrifugation and cultured in complete medium. Exosomes were isolated from cell supernatants by ultracentrifugation for cross-co-culture. After 48 h， migration and invasion abilities were assessed by scratch and Transwell assays. Sequencing was then performed on EEC-origin exosomal miRNA. ResultsThe model rats exhibited thin endometrium， along with reduced blood vessels， glandules， and pinopode numbers. BSHX improved endometrial morphology and increased pinopode numbers. MUC1， FoxO1， and HoxA10 expressions were downregulated in the model rats， while these parameters were upregulated after BSHX medium- and high-dose intervention. In the cell experiment， after exosome-free medicated serum intervention for 24 h， migration and invasion abilities were enhanced in the BSHX groups （P<0.01）. In EEC-origin exosomal miRNA sequencing， gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） analyses revealed enrichment in biological processes （gastrulation， neuronal differentiation， alongside cell development and regeneration）， involving the mitogen-activated protein kinase （MAPK）， FoxO1， Wnt， mammalian target of rapamycin （mTOR）， and tumor necrosis factor （TNF） signaling pathways. ConclusionBSHX promotes embryo implantation by improving endometrial receptivity via regulating exosomal miRNA. These findings provide potential targets for exosomal miRNA-based assisted reproductive strategies and a novel theoretical basis for infertility treatment by traditional Chinese medicine.

Abstract To promote the mental health of college students and build campus based psychological support system, the study explains that current applications of digital health interventions (DHIs) include independent forms based on a single technology or platform, as well as integrated forms that combine multiple digital technologies with traditional mental health services. These interventions have positive effects on alleviating negative emotions among college students, enhancing psychological resilience, improving quality of life, and ameliorating academic performance. However, differences still exist in the stability of intervention effects. Future development requires continuous improvement in institutional construction, ethical governance and capacity building.

Objective:To compare the effectiveness between the unilateral biportal endoscopy (UBE) and the bilateral biportal endoscopy (BBE) decompression in the treatment of two-level central lumbar spinal stenosis (LSS).Methods:From January 2022 to April 2024, the clinical data of 31 patients with two-level central LSS treated with UBE and BBE unilateral approach with bilateral decompression were retrospectively analyzed. There were 17 males and 14 females; the age ranged from 60 to 82 years, with a mean of (71.2±5.9) years. The operative segments were L 2-3 and L 3-4 in 2 cases, L 3-4 and L 4-5 in 29 cases. Among them, 15 cases were treated with UBE and the other 16 cases were treated with BBE. The age, gender, course of disease, operation time, intraoperative fluoroscopy frequency, ambulation time, hospitalization days, incision healing grade and surgical complications were compared between the two groups. Visual analogue scale (VAS) and Oswestry disability index (ODI) were used to assess the low back and leg pain degree and functional improvement situation before operation, 3 months after operation and at last follow-up. Imaging examinations were performed before and after operation to evaluate the height of intervertebral space, the rate of articular process preservation and the area of dural sac in the two groups. Measurement data with normal distribution were represented as mean±standard deviation( ± s), and the comparison between groups was conducted using the t-test; measurement data with skewed distribution were represented as (interquartile range) [ M( Q1, Q3)], inter-group comparisons were conducted using the two-sample rank sum test, and intra-group comparisons before and after surgery were conducted using the rank sum test for two related samples or the rank sum test for multiple related sample data. The count data were represented as cuses and percentages, and the comparison between groups was conducted using the Chi-square test or Fisher exact probability method. Results:Thirty-one patients were successfully operated and followed up for 6-18 months, with an average follow-up time of (11.4±3.2) months. There was no significant difference in age, gender, course of disease, ambulation time and hospitalization days between the two groups ( P>0.05). There were significant differences between UBE and BBE in fluoroscopy frequency [(4.2±0.7) vs (2.3±0.4)] and operation time [(118.2±12.8) min vs (72.3±5.6) min] ( P<0.001). Three months after operation and at last follow-up, the VAS scores and ODI were significantly lower than that befor the operation, and the dural sac area was significantly larger than that before the operation in the two groups ( P<0.001), but there was no significant difference in VAS, ODI and dural sac area before or after operation between the two groups ( P>0.05). There was no statistical difference in the intervertebral height between the two groups compared to their respective preoperative measurements( P>0.05). The rate of articular process preservation on the operated side was about 80% in both groups. There were no complications such as dural nerve injury and hemorrhage in both groups. One patient in the UBE group had incision infection, which was improved after symptomatic treatment. Conclusions:Both UBE and BBE can achieve satisfactory effectiveness in the treatment of two-level central LSS, and the clinical effectiveness is similar. BBE can improve the operation efficiency, shorten the surgical duration and reduce the fluoroscopy frequency, so it has more advantages in the treatment of two-level central LSS.

The mutual transformation and intertweaving of qi stagnation and blood stasis exist through the development and progression of pediatric pneumonia.Based on the pathogenic mechanism of qi stagnation and blood stasis obstructing the lungs and by analyzing the progression of pneumonia,this paper proposed that the traditional Chinese medicine(TCM)pathogenesis of pediatric pneumonia can be divided into three stages:early,middle,and late.In the early stage,external pathogens invade and then lead to lung qi stagnation and blockage.In the middle stage,qi stagnation and blood stasis block the lungs and then cause blood vessel stagnation.In the late stage,qi stagnation and blood stasis linger,and then result into the residual pathogens lurking in the lungs.Accordingly,the principles of treating pediatric pneumonia are proposed,i.e.,ventilating the lungs and dispelling pathogens,activating blood circulation and resolving stasis,relieving qi stagnation and unblocking blockage.The strategies for treating pediatric pneumonia in three stages are also introduced:for the early stage of pneumonia characterized by lung qi stagnation and blockage,the formula Yinqiao San combined with Wuhu Erchen Decoction are used for clearing lung-heat,resolving phlegm,and relieving wheezing;for the middle stage of pneumonia characterized by the intertweaving of qi stagnation and blood stasis,the formula Xuefu Zhuyu Decoction combined with Xuanfei Baidu Prescription are used for activating blood circulation and resolving stasis,ventilating the lungs and removing toxins,relieving qi stagnation and unblocking blockage;for the late stage of pneumonia characterized by qi stagnation and blood stasis lingering in the lungs,Qinghao Biejia Decoction and Qingfei Touxie Prescription are used by modification for clearing heat and nourishing yin,resolving blood stasis and relieving qi stagnation,nourishing yin and subduing yang.The approach to treating pediatric pneumonia from the pathogenic mechanism of qi stagnation and blood stasis obstructing the lung show significant efficacy on improving clinical symptoms in children with pediatric pneumonia and preventing the progression of pneumonia,and will provide reference to the clinical prevention and treatment of pediatric pneumonia.

Metastasis remains the primary cause of cancer-related mortality worldwide. Circulating tumor cells (CTCs) represent critical targets for metastasis prevention and treatment. Traditional Chinese medicine may prevent lung cancer metastasis through long-term intervention in CTC activity. Tiao-Shen-Zhi-Ai Formular (TSZAF) represents a Chinese medicine compound prescription utilized clinically for lung cancer treatment. This study combined three principal active ingredients from TSZAF into a novel TSZAF monomer combination (TSZAF mc) to investigate its anti-metastatic effects and mechanisms. TSZAF mc demonstrated significant inhibition of proliferation, migration, and invasion in CTC-TJH-01 and LLC cells, while inducing cellular apoptosis in vitro. Moreover, TSZAF mc substantially inhibited LLC cell growth and metastasis in vivo. Mechanistically, TAZSF mc significantly suppressed the Wnt/β-catenin signaling pathway and CXCL5 expression in lung cancer cells and tissues. Additionally, TAZSF mc notably reduced neutrophil infiltration in metastatic lesions. These findings indicate that TSZAF mc inhibits lung cancer growth and metastasis by suppressing the Wnt/β-catenin signaling pathway and reducing CXCL5 secretion, thereby decreasing neutrophil recruitment and infiltration. TSZAF mc demonstrates potential as an effective therapeutic agent for lung cancer metastasis.
Lung Neoplasms/genetics* ; Wnt Signaling Pathway/drug effects* ; Animals ; Humans ; Drugs, Chinese Herbal/pharmacology* ; Mice ; Neoplasm Metastasis/prevention & control* ; Cell Proliferation/drug effects* ; Cell Line, Tumor ; Neutrophil Infiltration/drug effects* ; Down-Regulation/drug effects* ; Cell Movement/drug effects* ; beta Catenin/genetics* ; Apoptosis/drug effects* ; Mice, Inbred C57BL ; Male ; Neoplastic Cells, Circulating/drug effects*

With the rapid development of competitive sports, the incidence of anterior cruciate ligament (ACL) injury is on the rise. Such injuries may shorten athletes′ career and lead to other long-term adverse consequences. Although athletes generally recover well after ACL reconstruction, many still struggle to return to their pre-injury performance levels. Advances in the understanding of ACL anatomy and injury mechanisms, along with the evolution of surgical techniques and rehabilitation methods, have provided more individualized and tailored options for athletes following ACL injuries. However, there is currently no consensus in China regarding surgical and rehabilitation strategies for competitive athletes aiming to return to sports after ACL injuries. To this end, the Sports Medicine Committee of the Chinese Research Hospital Association and the Editorial Board of the Chinese Journal of Trauma jointly formulated the Expert consensus on surgical treatment and rehabilitation for competitive sports athletes returning to sports after anterior cruciate ligament injury ( version 2025), and presented 14 recommendations covering surgical indications, preoperative rehabilitation, surgical timing, surgical strategies and postoperative rehabilitation strategies, aiming to improve the surgical treatment and rehabilitation system for ACL injuries in competitive athletes and facilitate their return to high-level sports performance after injury.

Pulmonary anthrax is an acute zoonotic infectious disease caused by Bacillus anthracis infecting the human body. Humans can be infected by coming into contact with diseased animals and their products or by consuming diseased animals. According to the different infection routes, it can be classified into cutaneous anthrax, pulmonary anthrax and intestinal anthrax, etc. Among them, cutaneous anthrax is the most common, while case reports of pulmonary anthrax are rare. This article analyzes the clinical data of a case of occupational pulmonary anthrax diagnosed by metagenomic next-generation sequencing (mNGS) and successfully treated, so as to provide a reference for the clinical treatment of pulmonary anthrax.

Objective:To investigate the clinical efficacy of open arthrolysis in the treatment of posttraumatic elbow stiffness.Methods:A retrospective analysis was conducted on the data of 407 patients with post-traumatic elbow stiffness treated by open arthrolysis surgery in Shandong Provincial Hospital from January 2010 to January 2024. The cohort included 303 males and 104 females, with a mean age of 38.98±10.90 years (range, 18-72 years) and mean body mass index (BMI) of 24.32±3.29 kg/m 2 (range, 17.91-33.41 kg/m 2). There were 230 patients with right-sided elbow stiffness, 159 patients with left-sided elbow stiffness, and 18 patients with bilateral elbow stiffness. Initial injuries included 21 patients of isolated elbow dislocation; 25 patients of soft tissue injury; and 361 patients of initial intra-articular elbow fractures, among which there were 200 patients of multiple fractures, 87 patients of single distal humerus fracture, 43 patients of single proximal ulna fracture, and 31 patients of single radial head fracture. Initial injuries were treated non-surgically in 69 cases and surgically in 338 cases, among which 177 cases were retained with internal fixation. There were 334 preoperative patients complicated with heterotopic ossification and 73 patients without heterotopic ossification, with 99 patients undergoing early release (stiffness duration <6 months) and 308 patients undergoing late release (stiffness duration ≥6 months). Record the range of motion (ROM) of the elbow joint, forearm rotational range (FRR), visual analogue scale (VAS), Mayo elbow performance score (MEPS), modified Broberg-Morrey score (MBS), Oxford elbow score (OES), and disability of arm, shoulder and hand (DASH) score before and after surgery, and conduct comparative analysis. Results:All patients were followed up for an average of 41.86±10.27 months (range, 13-119 months). At 12 months postoperatively, elbow ROM improved from preoperative 33.7°±26.5° to 101.2°±24.0°, elbow FRR improved from preoperative 101.4°±53.5° to 138.9°±38.7°, the MEPS increased from 60.1±14.7 to 91.5±10.1, the BMS increased from 57.5±12.8 to 83.7±11.0, the OES decreased from 31.6±7.3 to 16.0± 4.6, the DASH score decreased from 38.8±13.9 to 10.1±9.5, and the VAS decreased from 3.0±2.3 to 0.9±1.1, with all changes showing statistical significance ( P<0.05). In patients with preoperative heterotopic ossification, postoperative mean flexion range was 120.1°±15.5° and elbow ROM was 102.6°±23.4°. In patients without preoperative heterotopic ossification, postoperative mean flexion range was 113.9°±15.6° and elbow ROM was 93.4°±26.4°. Statistically significant differences were observed between the two groups in postoperative flexion range and flexion-extension ROM. There were no statistically significant differences in the postoperative above-mentioned indicators between early and late release patients ( P>0.05). The supination range and elbow FRR in patients with multiple fractures were lower than those in patients with distal humerus fractures and proximal ulna fractures; the DASH score in patients with multiple fractures was higher than that in patients with proximal ulna fractures and radial head fractures; the OES score in patients with multiple fractures was higher than that in patients with proximal ulna fractures, and all differences were statistically significant ( P<0.05). Among 407 patients, complications included new-onset postoperative ulnar neuropathy in 61 cases, new heterotopic ossification in 11 cases, recurrent heterotopic ossification in 96 cases, elbow instability in 6 cases, and superficial surgical site infection in 2 cases. Conclusions:Open arthrolysis is an effective treatment option for post-traumatic elbow stiffness. Patients with preoperative heterotopic ossification have a greater postoperative flexion range and elbow flexion-extension range of motion. The surgical timing exerts no significant influence on the ultimate functional outcome of treatment in patients with post-traumatic elbow stiffness. Patients with different initial fracture sites exhibited significant differences in postoperative functional outcomes, including supination, DASH scores, and OES.

The aim of this updated guideline is to provide comprehensive recommendations for the management of gout in patients with common comorbidities, such as chronic kidney disease(CKD), cardiovascular disease(CVD), diabetes, osteoarthritis(OA), and gastrointestinal disorders. This guideline was developed by a multidisciplinary expert panel consisting of specialists in endocrinology, rheumatology, nephrology, cardiology, gastroenterology, and methodology. The development process adhered to standard methodologies, including PICO(population, intervention, comparator, and outcomes) question deconstruction, systematic literature review, the Grading of Recommendations Assessment, Development and Evaluation(GRADE) for evidence and recommendation evaluation, Delphi voting, and expert consensus. The guideline presents 26 evidence-based recommendations addressing 7 clinical questions for patients with hyperuricemia and gout in the context of comorbidities. Key recommendations include the maintenance of strict serum urate targets, particularly for patients with CKD stage≥3, chronic gouty arthritis, and OA, in order to prevent disease progression. In patients with CVD or diabetes, intra-articular triamcinolone is preferred over systemic glucocorticoids. Prioritized anti-inflammatory treatments for patients with CKD, gastrointestinal diseases and OA are recommended. The guideline also introduces emerging therapies, such as interleukin-1 inhibitors and selective urate transport inhibitors, as potential treatment options for refractory cases. The update offers a comprehensive, patient-centered approach to managing gout, particularly in individuals with associated comorbidities. Multidisciplinary collaboration and emerging new treatments and evidence ensure the optimization of the recommendations.