1.Construction and characterization of recombinant human coagulation factor Ⅶ stable transfected cell lines
Xiaoxiao LI ; Jiabin CHEN ; Jiajun LIU ; Zhifei ZHANG ; Sen ZOU ; Lihua ZHU ; Zhaoyong YANG
Acta Universitatis Medicinalis Anhui 2026;61(1):16-22
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.
2.Mechanism of Embryo Implantation Promotion via Exosomal miRNA-mediated Communication Network at Maternal-fetal Interface Based on Bushen Huoxue Therapy
Pei GUO ; Jiajun LIU ; Hang ZHOU ; Zeyi GUO ; Yili WANG ; Linwen DENG ; Qian ZENG ; Jinzhu HUANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(14):317-327
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.
3.Analysis of the efficacy of modified electroconvulsive therapy in schizophrenia patients across different genders
Jiajun REN ; Yuting LI ; Tingting WANG ; Shuangshuang FENG ; Hongcheng XIE ; Junfan LIANG ; Hongli ZHANG ; Ziyuan LIN ; Bo XIANG ; Kezhi LIU ; Xuemei LIANG
Chinese Journal of Nervous and Mental Diseases 2025;51(2):89-94
Objective To Investigate the efficacy of modified electroconvulsive therapy(MECT)in patients with schizophrenia across different genders.Methods From May 2018 to August 2022,481 patients with schizophrenia were recruited from three psychiatric hospitals in Luzhou,Zigong,and Yibin.According to gender grouping,both groups received adjunctive MECT treatment for two consecutive weeks for a total of six treatments.The differences in positive and negative syndrome scale(PANSS)scores before and after treatment,UKU adverse reaction rating scale(UKU),and gastrointestinal symptom rating scale(GSRS)scores were compared between the two groups.Results After quality control,463 cases were followed up for analysis including 246 males and 217 females.Compared with pre-treatment,the total PANSS score and scores on each subscale were significantly reduced in both genders after treatment(P<0.001).When comparing the reduction rates between the groups,the male patients showed a higher reduction rate in negative symptoms than the female patients(31.24%±30.24%vs.25.80%±33.96%,P<0.05).However,there were no significant differences between the two groups in the reduction rates of the total score,positive symptoms,and general psychopathology(P>0.05).The comparison of adverse reactions showed that the frequency of other types of adverse reactions was higher in female patients than in male patients(47.47%vs.37.80%,P<0.05).However,no significant differences were observed in the adverse reactions related to the mental,neurological,autonomic nervous system,and gastrointestinal systems(P>0.05).Correlation analysis revealed that the reduction rate of the PANSS total score was positively correlated with smoking history(r=0.135,P=0.034)and alcohol history(r=0.160,P=0.012)in male patients,while the reduction rate of the PANSS total score was negatively correlated with the disease duration(r=-0.210,P=0.002)and positively correlated with the age of onset(r=0.145,P=0.032)in female patients.Conclusion MECT is significantly effective for both male and female patients with schizophrenia.Compared to female patients,MECT shows a more pronounced effect on negative symptoms in male patients.Additionally,the factors related to the efficacy of MECT differ between genders,indicating that it is necessary to consider the clinical characteristics of patients comprehensively when selecting an MECT treatment plan.
4.Research progress on influencing factors and countermeasures of scientific research outputs among medical personnel
Xin CHEN ; Liyan LONG ; Lei ZHANG ; Jiajun DU ; Tingfang LIU
Modern Hospital 2025;25(3):476-479
As the main body of scientific research and innovation in hospitals,timely understanding of medical person-nel's scientific research outputs and its influencing factors has practical guiding significance for hospital management to identify scientific research talents,formulate targeted training programs,customize relevant science and technology policies,etc.This ar-ticle will comprehensively elaborate the concept of scientific research outputs,evaluation methods and influencing factors of scien-tific research outputs among medical personnel.Meanwhile,the article will provide suggestions for influencing factors,in order to provide references for the hospital management in the future.
5.Mechanistic study of Yigan Fupi decoction-mediated repair of the intestinal barrier and reduction of IBS sensitivity through regulation of the PKA/PKC-CREB pathway
Yu CHEN ; Jiajun SHI ; Danting FU ; Qinqin YANG ; Rui FU ; Jiajie ZHU ; Mingjin ZHU ; Xinying LIU ; Mingxian CHEN
Acta Laboratorium Animalis Scientia Sinica 2025;33(4):512-521
Objective To study the therapeutic effects of Yigan Fupi decoction(YGFP)on irritable bowel syndrome(IBS)and its mechanism of action in repairing the intestinal barrier and reducing IBS sensitivity through the PKA/PKC-CREB pathway.Methods Baby rats separated from their mother were randomly divided into a model control(M)and a YGFP group,while baby rats without maternal separation were used as a normal control(N)group.The YGFP group was given YGFP for 4 weeks.Abdominal withdrawal reflux was used to evaluate intestinal sensitivity.Liquid chromatography tandem mass spectrometry and ELISA were used to detect bile acid metabolite concentrations and serum levels of interleukin(IL)-6 and CXCL1,respectively.HE staining was used to observe pathological changes in the colon,and Western Blot and immunohistochemistry were used to analyze the relative protein expression levels of PKA,PKC,CREB,5HT2AR,5-HT7R,ZO-1,and Claudin 1.Results Compared with the normal control group,the M group showed a significantly decreased visceral pain threshold,significantly increased levels of total bile acid metabolites,IL-6,and CXCL1,significantly increased relative expression of PKA,PKC,CREB,5HT2AR,and 5-HT7R,and significantly decreased relative expression of ZO-1 and Claudin 1.Compared with the M group,the YGFP group showed a significantly increased visceral pain threshold,significantly reduced levels of total bile acid metabolites,IL-6,and CXCL1,significantly reduced relative expression of PKA,PKC,CREB,5HT2AR,and 5-HT7R,and increased relative expression of ZO-1 and Claudin 1.Conclusions YGFP effectively improved IBS through a mechanism that may involve repair of the intestinal barrier and reduced sensitivity through the PKA/PKC-CREB pathway.
6.SRSF1 inhibits HBV replication by enhancing the stability of P53 in cell models
Jiajun LIU ; Shaoyuan LONG ; Jieli HU ; Jing CUI
Journal of Army Medical University 2025;47(13):1475-1483
Objective To investigate the effect and underlying mechanism of serine and arginine rich splicing factor 1(SRSF1)on the replication of hepatitis B virus(HBV).Methods The effects of SRSF1 on HBV replication were investigated in different HBV replicating cell models by Southern blotting,Northern blotting and ELISA.Quantitative PCR,luciferase reporter assay and chromatin immunoprecipitation(ChIP)were applied to determine the effects of SRSF1 on the activities of HBV core promoter/enhancer in HepG2 cells.The relationship between SRSF1 and P53 was explored with Western blotting and ubiquitination assay.The effects of P53 on HBV replication were verified in different HBV replicating cell models,and the role of P53 in SRSF1 inhibition of HBV was clarified.Results Overexpression of SRSF1 significantly inhibited HBV DNA and HBV RNA and reduced HBsAg and HBeAg secretion levels in a variety of HBV replicative cell models(P<0.0001).SRSF1 also inhibited the activity of HBV core promoter,although this inhibition was regulated by indirect mechanisms.In addition,SRSF1 enhanced P53 stability by protecting P53 from ubiquitination and subsequent proteasomal degradation.Meanwhile,the regulatory effect of overexpression or knockdown of P53 on HBV was validated in different cell models(P<0.0001).Conclusion Overexpression of splicing factor SRSF1 significantly inhibits HBV replication in a variety of cell models,and this inhibitory effect is mediated by its enhancement of P53 stability.
7.Development of a multimodal deep learning-based risk prediction model integrating clinical and radiomic features for short-term acute kidney injury following partial nephrectomy
Jiangting CHENG ; Jiayi XU ; Chenyang SHEN ; Guanwen YANG ; Yaohui LI ; Li LIU ; Jiajun WANG ; Xiaoyi HU ; Jianming GUO ; Hang WANG
Chinese Journal of Urology 2025;46(5):349-355
Objective:To develop and validate a deep learning-based multimodal model integrating clinical and radiomic features for predicting acute kidney injury(AKI)risk after partial nephrectomy.Methods:A retrospective analysis was conducted on 416 patients who underwent partial nephrectomy at Zhongshan Hospital,Fudan University from January 2023 to January 2025. The cohort included 100 AKI patients[defined by a ≥ 25% reduction in postoperative evaluated glomerular filtration rate(eGFR)within 48 hours sustained for >24 hours]and 316 non-AKI patients(1∶3 ratio,randomly matched with 16 additional cases for redundancy). Clinical and radiomic features were extracted from preoperative contrast-enhanced CT scans using PyRadiomics. Demographics included 259 males and 158 females,with a median age of 57(49,65)years,body mass index of(24.1 ± 3.3)kg/m2,preoperative eGFR of(88.5 ± 18.3)ml/(min·1.73 m2),postoperative eGFR(48-hour)of(76.0 ± 21.9)ml/(min·1.73 m2),Zhongshan Score(ZSscore)of 7.34 ± 2.01,and R.E.N.A.L. score of 7.50 ± 1.71. All tumors were T 1a stage. Patients were divided into training(n = 312)and test(n = 104)sets(3∶1 ratio). A clinical model was constructed via multivariate logistic regression,while radiomic and combined(clinical + radiomic)models utilized an artificial neural network(ANN)with 1 input layer,5 hidden layers,1 output layer,and 10 5 training epochs. Model performance was evaluated by using receiver operating characteristic(ROC)curves and area under the curve(AUC),and was compared to the Martini model. Feature contributions were interpreted via SHapley Additive exPlanations(SHAP). Results:In the test set,the results of multivariate logistic regression showed that patient’s weight,preoperative eGFR,R.E.N.A.L. score,surgical approach,and operation time were risk factors for AKI( P < 0.05). The AUC of the clinical feature prediction model constructed based on the above factors was 0.852(95% CI 0.775?0.929). In the test set,the AUC of the Martini model was 0.725(95% CI 0.565?0.791). The radiomic model,trained on 1 315 imaging features,achieved an AUC of 0.898(95% CI 0.804?0.993)with 94.2%(98/104)accuracy. The combined clinical and radiomic model,integrating 1 315 radiomic features and clinical features,demonstrated superior performance with an AUC of 0.946(95% CI 0.887?1.000)and 96.2%(100/104)accuracy,outperforming both the clinical model( P = 0.03)and the Martini model( P < 0.01). SHAP analysis identified the top five predictors in the combined model:ZSscore(SHAP value:0.78),long-run low gray-level emphasis(SHAP value:0.61),run-length non-uniformity(SHAP value:0.58),size-zone non-uniformity(SHAP value:0.46),and gray-level co-occurrence matrix joint energy(SHAP value:0.36). Conclusions:The deep learning-based multimodal model integrating clinical and radiomic features accurately predicts AKI risk after partial nephrectomy,offering a novel strategy for preoperative risk stratification and personalized intervention.
8.Exploration on the Mechanism of Danggui Sini Decoction in Treating Knee Osteoarthritis Based on Network Pharmacology and Experimental Validation
Mingzhan LIANG ; Yongju YANG ; Heshan YU ; Xuefeng GUAN ; Yuanhe FAN ; Jiajun LIU
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(10):32-40
Objective To explore the mechanism of Danggui Sini Decoction in treating knee osteoarthritis(KOA)by using network pharmacology,combined with in vitro experimental verification.Methods The active components of Danggui Sini Decoction were retrieved and screened through TCMSP,CNKI and PubMed databases,and their corresponding targets were predicted using the SwissTarget Prediction platform.KOA related targets were retrieved from GeneCards and OMIM databases.Intersection of drug targets and KOA targets was obtained,and key components,core targets,and their related biological mechanisms were identified through network topology analysis,GO and KEGG pathway enrichment analysis.Molecular docking was used to verify the degree of binding between key components and core targets.Danggui Sini Decoction containing serum was prepared,and an in vitro KOA model was constructed by inducing rat articular chondrocytes with lipopolysaccharide.The CCK-8 method was used to screen for the optimal concentration of drug containing serum intervention,fluorescent probes were used to detect intracellular ROS content,immunofluorescence was used to detect NF-κB p65 nuclear translocation,RT-qPCR was used to detect the expressions of IL-6,IL-1β and TNF-α mRNA,Western blot was used to detect the expression of Toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88),NF-κB p65 and p-NF-κB p65 proteins.Results The results of network pharmacology analysis indicated that the treatment of KOA with Danggui Sini Decoction may involve regulating Toll like receptors,MAPK,TNF,apoptosis and other inflammatory and aging signaling pathways,compounds such as quercetin,kaempferol and glycyrrhizin may play important roles.Core targets included IL6,IL1B,TNF,TLR4,NFKB1,JUN,MAPK1,RELA.In vitro experiments showed that compared with the blank group,the ROS content in chondrocytes of the model group significantly increased(P<0.01),NF-κB p65 protein was significantly nuclear translocation(P<0.01),and mRNA expressions of IL-6,IL-1β,TNF-α mRNA and protein expressions of TLR4,MyD88 and p-NF-κB p65 significantly increased(P<0.01);compared with the model group,the intervention of Danggui Sini Decoction containing serum could significantly reduce intracellular ROS content(P<0.05),inhibit NF-κB p65 nuclear translocation(P<0.01),and reduce the expression of inflammatory factor mRNA and related proteins(P<0.05,P<0.01).Conclusion Danggui Sini Decoction can significantly reduce the inflammatory response of rat articular chondrocytes induced by lipopolysaccharide and exert therapeutic effects on KOA.Its mechanism may be related to the inhibition of TLR4/MyD88/NF-κB pathway.
9.Clinical efficacy of intensive conservative treatment for acute aortic syndrome
Yinfan ZHU ; Lu DAI ; Haotian WU ; Yamin LI ; Dongjie LI ; Shipan WANG ; Jiajun LIANG ; Yan YAN ; Jianjun GAO ; Yeting LOU ; Zhenze TAO ; Yifan LU ; Zhiran YANG ; Jia LI ; Siji CHEN ; Chuang LIU ; Yazhe ZHANG ; Yuhong MI ; Haiyang LI ; Wenjian JIANG ; Hongjia ZHANG
Chinese Journal of Thoracic and Cardiovascular Surgery 2025;41(3):143-150
Objective:To evaluate the outcomes of intensive conservative treatment compared to conventional conservative treatment in patients with acute aortic syndrome(AAS).Methods:The study prospectively enrolled consecutive patients with AAS who were admitted to Beijing Anzhen Hospital, affiliated with Capital Medical University, and Beijing Dawanglu Emergency Rescue Hospital from January 2024 to December 2024. These patients with surgical contraindications or refused surgery for various reasons opted for conservative treatment. A total of 282 patients were included, and 15 patients with missing data or those who died without any treatment were excluded. Finally, 267 patients were enrolled, of whom 94 received intensive conservative treatment, and 173 received conventional conservative treatment, the inverse probability of treatment weighting (IPTW) was used to reduce the influence of confoundings. After adjusting of baseline datas via IPTW, the survival outcomes of the two groups were compared at 14 days, 30 days, and at the end of follow-up.Results:The results showed significant differences in acute phase survival rates between the enhanced conservative treatment group and the conventional conservative treatment group at 14 days(82.40%vs.53.20%, P<0.0001). Significant survival differences were also observed at 30 days and at 276-day mid-term follow-up (96.29% vs.51.60%, P<0.0001; 78.50% vs.48.50%, P<0.0001). In the subgroup analysis, for type A aortic dissection, the enhanced conservative treatment group had higher survival rates compared to the conventional conservative treatment group at 14, 30 and 276 days (63.46% vs.41.35%, P<0.05; 52.17% vs.37.90%, P<0.05; 50.00% vs. 31.97%, P<0.05). However, for type B aortic dissection, although the enhanced conservative treatment group had higher survival rates than the conventional conservative treatment group, no statistically significant differences were observed (96.29% vs. 80.00%, P=0.054; 95.65% vs.78.37%, P=0.067; 94.12% vs.74.20%, P=0.088). Conclusion:For patients diagnosed with AAS are forced to choose conservative treatment if emergency surgery is not possible in the first place, intensive conservative treatment strategies can significantly reduce the mortality in the acute phase compared with conventional conservative treatment. Mid-term follow-up, intensive conservative treatment still has a significant survival advantage.
10.Expert consensus on surgical treatment and rehabilitation for competitive sports athletes returning to sports after anterior cruciate ligament injury (version 2025)
Kai HUANG ; Lunhao BAI ; Qing BI ; Hong CHEN ; Jiwu CHEN ; Xuesong DAI ; Wenyong FEI ; Weili FU ; Zhizeng GAO ; Lin GUO ; Yinghui HUA ; Jingmin HUANG ; Suizhu HUANG ; Xuan HUANG ; Jian LI ; Qiang LI ; Shuzhen LI ; Yanlin LI ; Yunxia LI ; Zhong LI ; Ning LIU ; Yuqiang LIU ; Wei LU ; Hongbin LYU ; Haile PAN ; Xiaoyun PAN ; Chao QI ; Weiliang SHEN ; Luning SUN ; Jin TANG ; Zimin WANG ; Bide WANG ; Ru WANG ; Shaobai WANG ; Licheng WEI ; Weidong XU ; Yongsheng XU ; Jizhou YANG ; Liang YANG ; Rui YANG ; Hongbo YOU ; Tengbo YU ; Jiakuo YU ; Bing YUE ; Hua ZHANG ; Hui ZHANG ; Qingsong ZHANG ; Xintao ZHANG ; Jiajun ZHAO ; Lilian ZHAO ; Qichun ZHAO ; Song ZHAO ; Jiapeng ZHENG ; Jiang ZHENG ; Zhi ZHENG ; Jingbin ZHOU ; Jinzhong ZHAO
Chinese Journal of Trauma 2025;41(4):325-338
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.

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