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.Shentong Zhuyutang Regulates SIRT1/Nrf2 Pathway to Ameliorate Intervertebral Disc Degeneration in Rats
Jiajun HUANG ; Diyou WU ; Guangyi TAO ; Yu ZHAO ; Junqing HUANG ; Bin YANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):29-39
ObjectiveTo study the effect and mechanism of Shentong Zhuyutang in treating intervertebral disc degeneration (IDD) in rats. MethodsIn the cell experiment, male rats were administrated with normal saline or low-, medium-, and high-dose (3.38, 6.75,13.5 g·kg-1, respectively) Shentong Zhuyutang by gavage, respectively, and serum samples were collected after 7 days of continuous administration. Another 10 male rats were selected for the isolation of nucleus pulposus cells. The cell model of IDD was established by treatment with interleukin (IL)-1β. The modeled cells were then treated with Shentong Zhuyutang-containing serum and the ferroptosis inhibitor ferrostatin-1 (Fer-1), respectively, to investigate the effects of Shentong Zhuyutang-containing serum on the proliferation and ferroptosis of nucleus pulposus cells. To study the role of silent information regulator 1 (SIRT1)/nuclear factor erythroid 2-related factor 2 (Nrf2) in the regulation of ferroptosis in nucleus pulposus cells by Shentong Zhuyutang-containing serum, this study treated the cells with the SIRT1 inhibitor Ex 527 and the Nrf2 inhibitor ML385, respectively, in addition to the treatment with IL-1β and high-dose Shentong Zhuyutang-containing serum. The cell-counting kit-8 (CCK-8) assay and EdU staining were employed to measure the cell viability and proliferation, respectively. The Fe2+, glutathione (GSH), and malondiadehyde (MDA) levels were measured by colorimetric assay. Western blot was employed to determine the protein levels of glutathione peroxidase 4 (GPX4), acyl-CoA synthetase long-chain family 4 (ACSL4), Collagen Ⅱ, Aggrecan, SIRT1, and Nrf2. Immunofluorescence was used detect SIRT1 expression. In the animal experiment, male rats were treated with anulus puncture for the modeling of IDD. Rats were randomly assigned into sham operation, model, Shentong Zhuyutang-containing serum (13.5 g·kg-1), and positive control (nimesulide dispersible tablets, 0.18 mg·kg-1) groups. Rats in the drug intervention groups were administrated with corresponding agents at 1 mL·kg-1, and those in the sham operation and model groups were administrated with equal volumes of normal saline, once daily for 28 consecutive days. At the end of the last administration, the histopathological changes in the intervertebral discs of rats were observed by hematoxylin-eosin staining and scored by the Masuda method. Western blot was employed to determine the protein levels of SIRT1, Nrf2, GPX4, and Collagen Ⅱ in the nucleus pulposus tissue. ResultsCompared with the control group, the IL-1β group of nucleus pulposus cells showed elevated levels of Fe2+, MDA, and ACSL4 (P<0.05), decreased cell viability, lowered GSH level, and down-regulated protein levels of GPX4, Collagen Ⅱ, and Aggrecan (P<0.05). Shentong Zhuyutang-containing serum and Fer-1 reversed the effects of IL-1β on the viability and ferroptosis of nucleus pulposus cells and up-regulated the protein levels of Collagen Ⅱ and Aggrecan in nucleus pulposus cells (P<0.05). Compared with the control group, the IL-1β group showcased down-regulated expression of Sirt1 and Nrf2 in nucleus pulposus cells (P<0.05). Compared with the IL-1β group, the high-dose Shentong Zhuyutang-containing serum+IL-1β group showed up-regulated expression of SIRT1 and Nrf2 in nucleus pulposus cells (P<0.05). Compared with the high-dose Shentong Zhuyutang-containing serum+IL-1β group, the ML385 group showed down-regulated protein levels of Nrf2 and GPX4, lowered GSH level, and elevated Fe2+ and MDA levels (P<0.05). In addition, the Ex 527 group showed down-regulated protein levels of SIRT1, Nrf2, and GPX4 (P<0.05). The results of the animal experiment showed that compared with the sham operation group, the model group had severe degeneration of the intervertebral disc tissue with increased pathological score, up-regulated protein level of ACSL4 (P<0.05), and down-regulated protein levels of SIRT1, Nrf2, GPX4, and Collagen Ⅱ (P<0.05). Compared with the model group, the Shentong Zhuyutang group showed alleviated IDD with declined pathological score, down-regulated protein level of ACSL4 (P<0.05), and up-regulated protein levels of SIRT1, Nrf2, GPX4, and Collagen Ⅱ (P<0.05). ConclusionShentong Zhuyutang may activate the SIRT1/Nrf2 signaling pathway to inhibit the ferroptosis of nucleus pulposus cells, thereby delaying the process of IDD in rats.
3.Assessing distribution characteristics and clinical significance of vertebral fractures in patients with osteoporosis based on whole spine MRI
Jiajun ZHOU ; Fei MA ; Yebo LENG ; Shicai XU ; Baoqiang HE ; Yang LI ; Yehui LIAO ; Qiang TANG ; Chao TANG ; Qing WANG ; Dejun ZHONG
Chinese Journal of Tissue Engineering Research 2025;29(9):1883-1889
BACKGROUND:Osteoporotic vertebral fractures are the most common complication in patients with osteoporosis.As a new imaging technique,spine magnetic resonance imaging(MRI)is much more sensitive than X-ray film in the diagnosis of osteoporotic vertebral fractures.However,total spine MRI is costly and takes a long time to scan.Therefore,there is no consensus on whether all patients with osteoporotic vertebral fractures need to undergo total spine MRI scan and which patients need to undergo total spine MRI. OBJECTIVE:To analyze the distribution characteristics of vertebral fractures and explore their clinical significance by observing the whole spine MRI data of osteoporotic vertebral fractures patients. METHODS:Data of cases and MRI images of all patients diagnosed with fresh osteoporotic vertebral fractures who visited the Department of Orthopedics,Affiliated Hospital of Southwest Medical University from August 2018 to September 2022 were retrospectively analyzed.903 patients were included in the study based on inclusion and exclusion criteria.General information(age,gender,and body mass index),medical history characteristics(duration of illness,history of trauma surgery,percussion pain area,and pain score)were collected.The characteristics of vertebral fractures were analyzed through whole spine magnetic resonance imaging.Firstly,based on the number of vertebral fractures in patients,they were divided into the single vertebral fracture group(484 cases)and the multi-vertebral fracture group(419 cases),and the differences were analyzed between the two groups.Then,based on whether the farthest interval between the fractured vertebrae was greater than or equal to 5,the multi vertebral fracture group was further divided into two subgroups.Among them,Group A(the farthest interval between the fractured vertebrae was less than 5)contained 306 cases;Group B(with the farthest interval between fractured vertebral bodies greater than 5)included 113 cases.The differences were analyzed between two subgroups. RESULTS AND CONCLUSION:(1)Among 903 patients,419 patients(46.4%)had more than two fractured vertebrae.There were 654 patients(72.4%)with thoracolumbar fractures,and 54 patients(6%)with fractures in the thoracic plus lumbar region and the entire thoracic to lumbar region.In group B,96.5%of patients had multiregional percussion pain.(2)Compared with the patients in the single vertebral fracture group and the multi-vertebral fracture group,there were significant differences in bone mineral density,whether the medical history was greater than or equal to 1 month,the history of low energy injury,and the distribution and number of axial percussion pain areas in the spine during physical examination between the two groups(P<0.05).Age,gender,body mass index,whether there was underlying disease,pain visual analog scale score,whether there was a history of elderly thoracolumbar fracture,and whether there was a history of thoracolumbar surgery,and the number of fractured vertebrae had no statistical significance(P>0.05).(3)There were statistically significant differences between the Groups A and B in bone mineral density,the distribution and quantity of percussion pain area,and the history of low energy injury(P<0.05).There were no significant differences in age,gender,history of old fractures,visual analog scale score,body mass index,whether the medical history was longer than or equal to 1 month,history of underlying diseases,and history of thoracolumbar surgery between the two groups(P>0.05).(4)Patients with multiple low-energy trauma history,history of more than 1 month,multiple percussion pain,and the lower bone mineral density should be alert to the occurrence of multiple vertebral fracture and jump fracture.We recommend the whole spinal MRI for these patients.
4.The correlation analysis between sacral slope and the morphological characteristics of intervertebral disc, paraspinal muscle and pedicle in patients with degenerative lumbar spondylolisthesis
Yang LI ; Yebo LENG ; Chao TANG ; Shicai XU ; Baoqiang HE ; Jiajun ZHOU ; Yehui LIAO ; Qiang TANG ; Qing WANG ; Dejun ZHONG
Chinese Journal of Surgery 2025;63(9):806-813
Objective:To analyzed the correlation between the sacral slope (SS) and the slip degree of vertebral body, the morphological parameters of intervertebral disc, paraspinal muscle and pedicle in patient with degenerative lumbar spondylolisthesis (DLS).Methods:A retrospective cohort study was conducted. One hundred and forty patients with L 4-5 degenerative spondylolisthesis who visited the Department of Orthopedics, the Affiliated Hospital of Southwest Medical University from July 2018 to October 2022 were included. There were 32 males and 108 females, with an age of (62.6±10.0) years (range: 42 to 87 years) and the body mass index was (23.8±2.8) kg/m 2(range: 14.7 to 28.7 kg/m 2). General information and imaging data (lumbar X-ray, CT three-dimensional reconstruction, MRI images) of the patients were collected. SS and the slip degree (SD) of the L 4 vertebral body were measured on lateral lumbar radiographs. Based on Roussouly classification, 140 patients with DLS were divided into a low SS group (SS<35°), a medium SS group (35°≤SS<45°), and a high SS group (SS≥45°). The morphological parameters of the L 4 pedicle of the patient were measured on the lumbar CT three-dimensional reconstruction images, including pedicle length (PL), pedicle width (PW), pedicle screw trajectory length (PSTL), pedicle camber angle(PCA) and pedicle height (PH). The morphological parameters of the paraspinal muscles (psoas major, multifidus and erector spinae) were measured on MRI images, including total cross-sectional muscle area (TCSA), functional cross-sectional muscle area (FCSA), and the ratio of functional cross-sectional muscle area (FCSA/TCSA) was calculated. The degree of degeneration of the L 4-5 disc was assessed on MRI images. The slip degree, morphological parameters of the pedicle, intervertebral disc, and paraspinal muscles among the groups were compared and their correlation was analyzed. The comparison of quantitative data was conducted using one-way analysis of variance, and the post hoc analysis for pairwise comparisons among groups was performed using the least significant difference method. The comparison of unordered categorical data was carried out using the χ2 test, and the comparison of ordered categorical data was conducted using the Kruskal-Wallis rank sum test. Pearson correlation analysis was used to test the correlation. Results:Among the 140 patients, there were 70 in the low SS group, 48 in the medium SS group, and 22 in the high SS group. The SD and PL of the low SS group were the lowest ((18.6±4.2)%, (14.0±1.0) mm), those of the medium SS group were slightly higher ((21.0±3.6)%, (15.7±0.9) mm), and those of the high SS group were the highest ((23.7±2.8)%, (16.2±0.7) mm), with statistically significant differences among the groups (all P<0.01). The PW, PH and PCA of the low SS group were the highest ((11.0±1.1) mm, (11.0±1.0) mm, 22.9°±1.1°), those of the medium SS group were slightly lower ((10.2±1.1) mm, (10.4±0.9) mm, 21.6°±1.3°), and those of the high SS group were the lowest ((9.6±0.9) mm, (9.4±0.7) mm, 18.7°±1.5°), with statistically significant differences among the groups (all P<0.05). The FSCA/TCSA of the psoas major, erector spinae and multifidus muscles in the low SS group were the highest ((93.1±4.0)% and (77.6±6.4) %), next were those in the medium SS group ((90.9±3.5)% and (74.9±6.1)%), and those in the high SS group were the lowest ((88.6±4.4)% and (71.2±7.4) %), with statistically significant differences among the different groups (all P<0.05). Pearson correlation analysis showed that SS was positively correlated with SD( r=0.557, P<0.01); SS was negatively correlated with the FSCA/TCSA of the paraspinal muscles (psoas major, erector spinae+multifidus) ( r=-0.393,-0.394,both P<0.01); SS was positively correlated with PL and PSTL ( r=0.706, P<0.01; r=0.264, P<0.01), and negatively correlated with PW, PCA and PH ( r=-0.469,-0.669,-0.549, all P<0.01). The Pfirrmann grade of intervertebral disc degeneration in the low SS group was better than that in the medium SS group and the high SS group( H=23.128, P<0.01). Conclusions:In DLS patients, as SS increases, the SD of the L 4 vertebral body gradually increases, the proportion of paraspinal muscle functional area gradually decreases, the degree of intervertebral disc degeneration gradually worsens, the pedicle gradually becomes slender, and the pedicle camber angle gradually decreases. There is a correlation between SS and the degree of vertebral slippage, as well as morphological parameters of pedicle, intervertebral disc, and paraspinal muscles.
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.Transcutaneous bilirubin curves in healthy neonates based on multicenter remote monitoring data
Bi ZE ; Xiaoyue DONG ; Jin WANG ; Chuan NIE ; Jiajun ZHU ; Fang GUO ; Falin XU ; Chunhui YANG ; Bizhen SHI ; Zhankui LI ; Xinhua ZHANG ; Jing LI ; Bin YI ; Xiuying TIAN ; Lejia ZHANG ; Jun TANG ; Xinlin HOU ; Jiahua XU ; Guoying HUANG ; Shuping HAN ; Wenhao ZHOU
Chinese Journal of Pediatrics 2025;63(12):1318-1324
Objective:To establish 30-day of age transcutaneous bilirubin (TcB) reference curves for healthy neonates, and to investigate regional variations in bilirubin dynamics.Methods:A multicenter retrospective cohort study was conducted. A total of 220 950 healthy neonates born at a gestational age of 35-<42 weeks, with a birth weight ≥2 000 g, who did not receive phototherapy within 60 h after birth were recruited. All of them underwent remote TcB monitoring using the Bilibaby remote jaundice monitoring system between August 1 st, 2020 and December 31 st, 2024 in 426 hospitals. TcB data were collected within the period from birth to 30-day of age. The P40, P75, and P95 of TcB values were calculated, and dynamic TcB curves for 30-day of age were constructed. Patterns of bilirubin change, rates of change, and transition outcomes were described. Regional comparisons between South and North were conducted using linear mixed-effects models for TcB trajectories and Pearson′s chi-square test for outcome differences. Results:A total of 220 950 neonates were included, of whom 101 711 (46.03%) were female. Gestational age at birth was (38.75±1.12) weeks, and birth weight was (3 272±417) g. TcB levels increased rapidly within 3-day of age, peaked at 4-6-day of age, with peak values at P40, P75, and P95 of 200.6, 239.7 and 275.4 μmol/L (11.8, 14.1 and 16.2 mg/dl), respectively. TcB levels gradually declined thereafter and stabilized after 13-day of age, with values at P40, P75, and P95 fluctuating between 147.9-159.8, 190.4-200.6, and 231.2-239.7 μmol/L (8.7-9.4, 11.2-11.8, 13.6-14.1 mg/dl), respectively. Notably, among neonates categorized as low-or low-intermediate-risk within 3-day of age, 6 700 (12.76%) progressed to intermediate-high or high risk between 4 and 30 days of age. Before 13-day of age, TcB levels in the southern regions were consistently higher than those in the northern regions ( P=0.039); from 14 to 30 days of age, the overall TcB levels had no statistically difference, but the temporal changes in TcB still showed regional differences (degrees of freedom=3, all interaction P<0.05). Among neonates classified as low-or low-intermediate risk within 3-day of age, 25 326 were from southern regions, of whom 4 254 (16.80%) progressed to intermediate-high or high risk between 4 and 30 days of age. In northern regions, 27 193 neonates were classified as low-or low-intermediate risk within 3-day of age, among whom 2 446 (8.99%) progressed to intermediate-high or high risk. The risk progression between the 2 regions had statistically difference ( χ2=716.49, P<0.001). Conclusions:A TcB percentile curve for neonates within 30-day of age was established, revealing that both the overall TcB level and its temporal trend were higher in southern than in northern newborns. These findings provide baseline data to support continuous management of neonatal jaundice.
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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