BACKGROUND:Formononetin demonstrates potent anti-inflammatory and antioxidant abilities.However,its protective effect on nucleus pulposus mesenchymal stem cells is not yet clear.OBJECTIVE:To investigate the effect and mechanism of formononetin on nucleus pulposus mesenchymal stem cells under an inflammatory microenvironment.METHODS:(1)Primary nucleus pulposus mesenchymal stem cells were isolated from the intervertebral discs of SD rats,and flow cytometry was performed to identify the surface markers of mesenchymal stem cells.(2)The CCK-8 assay was employed to evaluate the impact of lipopolysaccharide and formononetin on the proliferation viability of nucleus pulposus mesenchymal stem cells,aiming to determine the appropriate concentration of formononetin for subsequent cell treatments.(3)An inflammatory microenvironment was simulated by adding 5 μg/mL lipopolysaccharide to the DMEM/F-12 culture medium,and nucleus pulposus mesenchymal stem cells were treated with different concentrations of formononetin for 24 hours.Levels of inflammation markers were detected using western blot assay,real-time quantitative PCR,and immunofluorescence.Western blot assay was conducted to measure the protein levels of the nuclear factor kappa B signaling pathway.RESULTS AND CONCLUSION:(1)The nucleus pulposus mesenchymal stem cells cultured in adherent wall were shuttle-shaped with good growth status.The results of flow cytometry showed that the surface markers of mesenchymal stem cells were positive for CD29,CD44,and CD90,and the surface markers of hematopoietic stem cells were negative for CD34 and CD45.(2)The treatment with formononetin at 12.5,25,50,100,and 200 μmol/L concentrations for 24 hours had no significant proliferation inhibitory effect on nucleus pulposus mesenchymal stem cells.Compared with the lipopolysaccharide group,the cell viability of nucleus pulposus mesenchymal stem cells treated with 12.5,25,and 50 μmol/L formononetin for 24 hours was significantly increased,so formononetin at 12.5,25,and 50 μmol/L concentrations was subsequently selected as the low,medium,and high concentrations for treating nucleus pulposus mesenchymal stem cells.(3)Compared with the lipopolysaccharide group,the protein and mRNA expressions of matrix metalloproteinase-3,matrix metalloproteinase-13,and tumor necrosis factor-α in nucleus pulposus mesenchymal stem cells in the low,medium,and high concentrations of formononetin groups were significantly decreased(P＜0.05)in a dose-dependent manner.(4)Compared with the lipopolysaccharide group,the expressions of phosphorylated nuclear factor kappa B and phosphorylated nuclear factor kappa B inhibitor protein in nucleus pulposus mesenchymal stem cells in the low,medium,and high concentrations of formononetin groups were significantly decreased(P＜0.05)in a dose-dependent manner.The above results suggest that formononetin may attenuate lipopolysaccharide-induced inflammation in rat nucleus pulposus mesenchymal stem cells by inhibiting the activation of the nuclear factor kappa B signaling pathway.

Objective:To collect registered clinical research plans on TCM characteristic therapies for treating cervical spondylosis; To explore their research registration status; To provide references for future clinical trial registration and implementation.Methods:Clinical research on TCM characteristic therapies for treating cervical spondylosis was retrieved from ChiCTR, ITMCTR and Clinical Trials. gov from the establishment of the databases to July 1, 2024. Excel 2019 was used to conduct descriptive statistics on registration time, registration area and institution, funding source, research type and design scheme, research participation center and sample size, cervical spondylosis type, intervention measures, outcome indicators, reporting quality, research openness and methodological application.Results:A total of 138 clinical trials for the TCM treatment of cervical spondylosis were included, of which 136 were registered by domestic researchers in 22 provincial-level administrative regions. The top three in terms of registration numbers were Shanghai, Guangdong Province, and Beijing. Additionally, 2 were registered by foreign researchers in Egypt and Malaysia. The main sources of funding were 50 local finances, followed by 26 hospital subsidies and 18 national finances. The intervention research accounted for the largest proportion of research types, with 123 items (89.13%). The research center mainly focused on single center studies (98 projects). Most randomized controlled trials (115 trials) described randomization methods, while a small number of randomized controlled trials (50 trials) indicated blinding. The intervention measures were mostly combined with TCM therapy, and the outcome indicators were mainly efficacy indicators, with fewer safety indicators.Conclusions:At present, clinical trial registrations for TCM treatment of cervical spondylosis are increasing, but issues remain, such as poor study design, uneven distribution, and incomplete information. It is recommended to refine registration details, optimize study protocols, and promote high-quality clinical research.

BACKGROUND:Studies have shown that upregulation of heme oxygenase-1 expression enhances cellular antioxidant and anti-apoptotic abilities.However,the effects of upregulating heme oxygenase-1 expression on the proliferation and differentiation of neural stem cells under oxidative stress conditions remain unclear.OBJECTIVE:To investigate the influence of heme oxygenase-1 overexpression on the survival and differentiation capacity of neural stem cells under oxidative stress conditions.METHODS:(1)Mouse primary neural stem cells were isolated and cultured from newborn Balb/c mice.Immunofluorescence was used to detect the neural stem cell marker Nestin.(2)Lentivirus was used to infect neural stem cells to induce heme oxygenase-1 overexpression.Flow cytometry was used to assess green fluorescent protein fluorescence.Western blot assay was performed to detect the expression levels of heme oxygenase-1.(3)H2O2 was added to the lentivirus-infected neural stem cell culture medium to simulate the oxidative stress microenvironment after spinal cord injury.Effects of heme oxygenase-1 overexpression on neural stem cell proliferation and apoptosis levels were analyzed using cell proliferation assay kits,cell apoptosis assay kits,and TUNEL staining kits.(4)The levels of lipid oxidation markers malondialdehyde,catalase,superoxide dismutase,and glutathione peroxidase were detected using assay kits.(5)Flow cytometry was used to measure intracellular reactive oxygen species levels,and neural stem cell differentiation into astrocytes and neurons.(6)The effect of heme oxygenase-1 overexpression on neuronal axon growth during neural stem cell differentiation was observed under optical and fluorescence microscopes.RESULTS AND CONCLUSION:(1)Mouse neural stem cells exhibited stable morphology,good growth status,and high expression of Nestin as detected by immunofluorescence.(2)Western blot analysis showed that the overexpression of heme oxygenase-1 in the overexpression group was significantly higher than that in the empty carrier control group.Flow cytometry was used to detect the expression of green fluorescent protein in the neural stem cells of the heme oxygenase-1 overexpression group and empty vector control group.(3)Overexpression of heme oxygenase-1 maintained the proliferative activity of neural stem cells and significantly reduced the number of apoptotic cells under oxidative stress conditions.(4)Overexpression of heme oxygenase-1 inhibited lipid peroxidation of neural stem cells under oxidative stress microenvironment,enhanced the expression of enzymes related to maintaining the oxidative-reductive balance,and significantly reduced intracellular reactive oxygen species levels.(5)Overexpression of heme oxygenase-1 promoted the differentiation of neural stem cells into neurons and reduced differentiation into astrocytes.(6)The heme oxygenase-1 overexpression group exhibited longer axons,and more intercellular connections.The above results indicate that overexpression of heme oxygenase-1 can alleviate oxidative damage of H2O2-induced neural stem cells,reduce neural stem cell apoptosis,promote proliferation,and facilitate differentiation of neural stem cells into neurons.

BACKGROUND:Formononetin demonstrates potent anti-inflammatory and antioxidant abilities.However,its protective effect on nucleus pulposus mesenchymal stem cells is not yet clear.OBJECTIVE:To investigate the effect and mechanism of formononetin on nucleus pulposus mesenchymal stem cells under an inflammatory microenvironment.METHODS:(1)Primary nucleus pulposus mesenchymal stem cells were isolated from the intervertebral discs of SD rats,and flow cytometry was performed to identify the surface markers of mesenchymal stem cells.(2)The CCK-8 assay was employed to evaluate the impact of lipopolysaccharide and formononetin on the proliferation viability of nucleus pulposus mesenchymal stem cells,aiming to determine the appropriate concentration of formononetin for subsequent cell treatments.(3)An inflammatory microenvironment was simulated by adding 5 μg/mL lipopolysaccharide to the DMEM/F-12 culture medium,and nucleus pulposus mesenchymal stem cells were treated with different concentrations of formononetin for 24 hours.Levels of inflammation markers were detected using western blot assay,real-time quantitative PCR,and immunofluorescence.Western blot assay was conducted to measure the protein levels of the nuclear factor kappa B signaling pathway.RESULTS AND CONCLUSION:(1)The nucleus pulposus mesenchymal stem cells cultured in adherent wall were shuttle-shaped with good growth status.The results of flow cytometry showed that the surface markers of mesenchymal stem cells were positive for CD29,CD44,and CD90,and the surface markers of hematopoietic stem cells were negative for CD34 and CD45.(2)The treatment with formononetin at 12.5,25,50,100,and 200 μmol/L concentrations for 24 hours had no significant proliferation inhibitory effect on nucleus pulposus mesenchymal stem cells.Compared with the lipopolysaccharide group,the cell viability of nucleus pulposus mesenchymal stem cells treated with 12.5,25,and 50 μmol/L formononetin for 24 hours was significantly increased,so formononetin at 12.5,25,and 50 μmol/L concentrations was subsequently selected as the low,medium,and high concentrations for treating nucleus pulposus mesenchymal stem cells.(3)Compared with the lipopolysaccharide group,the protein and mRNA expressions of matrix metalloproteinase-3,matrix metalloproteinase-13,and tumor necrosis factor-α in nucleus pulposus mesenchymal stem cells in the low,medium,and high concentrations of formononetin groups were significantly decreased(P＜0.05)in a dose-dependent manner.(4)Compared with the lipopolysaccharide group,the expressions of phosphorylated nuclear factor kappa B and phosphorylated nuclear factor kappa B inhibitor protein in nucleus pulposus mesenchymal stem cells in the low,medium,and high concentrations of formononetin groups were significantly decreased(P＜0.05)in a dose-dependent manner.The above results suggest that formononetin may attenuate lipopolysaccharide-induced inflammation in rat nucleus pulposus mesenchymal stem cells by inhibiting the activation of the nuclear factor kappa B signaling pathway.

BACKGROUND:Studies have shown that upregulation of heme oxygenase-1 expression enhances cellular antioxidant and anti-apoptotic abilities.However,the effects of upregulating heme oxygenase-1 expression on the proliferation and differentiation of neural stem cells under oxidative stress conditions remain unclear.OBJECTIVE:To investigate the influence of heme oxygenase-1 overexpression on the survival and differentiation capacity of neural stem cells under oxidative stress conditions.METHODS:(1)Mouse primary neural stem cells were isolated and cultured from newborn Balb/c mice.Immunofluorescence was used to detect the neural stem cell marker Nestin.(2)Lentivirus was used to infect neural stem cells to induce heme oxygenase-1 overexpression.Flow cytometry was used to assess green fluorescent protein fluorescence.Western blot assay was performed to detect the expression levels of heme oxygenase-1.(3)H2O2 was added to the lentivirus-infected neural stem cell culture medium to simulate the oxidative stress microenvironment after spinal cord injury.Effects of heme oxygenase-1 overexpression on neural stem cell proliferation and apoptosis levels were analyzed using cell proliferation assay kits,cell apoptosis assay kits,and TUNEL staining kits.(4)The levels of lipid oxidation markers malondialdehyde,catalase,superoxide dismutase,and glutathione peroxidase were detected using assay kits.(5)Flow cytometry was used to measure intracellular reactive oxygen species levels,and neural stem cell differentiation into astrocytes and neurons.(6)The effect of heme oxygenase-1 overexpression on neuronal axon growth during neural stem cell differentiation was observed under optical and fluorescence microscopes.RESULTS AND CONCLUSION:(1)Mouse neural stem cells exhibited stable morphology,good growth status,and high expression of Nestin as detected by immunofluorescence.(2)Western blot analysis showed that the overexpression of heme oxygenase-1 in the overexpression group was significantly higher than that in the empty carrier control group.Flow cytometry was used to detect the expression of green fluorescent protein in the neural stem cells of the heme oxygenase-1 overexpression group and empty vector control group.(3)Overexpression of heme oxygenase-1 maintained the proliferative activity of neural stem cells and significantly reduced the number of apoptotic cells under oxidative stress conditions.(4)Overexpression of heme oxygenase-1 inhibited lipid peroxidation of neural stem cells under oxidative stress microenvironment,enhanced the expression of enzymes related to maintaining the oxidative-reductive balance,and significantly reduced intracellular reactive oxygen species levels.(5)Overexpression of heme oxygenase-1 promoted the differentiation of neural stem cells into neurons and reduced differentiation into astrocytes.(6)The heme oxygenase-1 overexpression group exhibited longer axons,and more intercellular connections.The above results indicate that overexpression of heme oxygenase-1 can alleviate oxidative damage of H2O2-induced neural stem cells,reduce neural stem cell apoptosis,promote proliferation,and facilitate differentiation of neural stem cells into neurons.

Objective To investigate the influencing factors associated with bleeding complications in patients with obstructive jaundice treated with percutaneous transhepatic cholangial drainage(PTCD).Methods Clinical data of 1 042 patients with obstructive jaundice,who received PTCD at the Affiliated Hospital of Guizhou Medical University,the Xiangya Second Hospital of Central South University,and the Affiliated Cancer Hospital of Guizhou Medical University of China between January 2015 and January 2021,were collected.The risk factors related to PTCD bleeding complications were retrospective analyzed.Results The location where the drainage tube forming loop had a statistically significant effect on PTCD bleeding complications(P＜0.01).Compared with the loop-forming within the common bile duct,the loop-forming within the left and right hepatic duct would increase the risk of postoperative bleeding by 155.6％(OR=2.556,95％CI:1.251-5.225),the loop-forming within the lower order branch of the left and right hepatic duct would increase the risk of postoperative bleeding by 414.4％(OR=5.144,95％CI:2.618-10.106).The difference in the risk degree of postoperative bleeding between different drainage ways after successful puncturing was statistically significant(P＜0.05).Compared with the external drainage method,internal-external joint drainage method would increase the risk degree of postoperative bleeding by 159.1％(OR=2.591,95％CI:1.102-6.091).Preoperative platelet count and preoperative total bilirubin level were the independent risk factors for bleeding complications of PTCD(P＜0.05).For each unit increase in preoperative platelet count,the probability of developing postoperative bleeding complications would decrease by 0.2％(OR=0.998,95％CI:0.995-1.000),and a preoperative platelet count level＜228 ×109/L would have an impact on the postoperative bleeding.For each unit increase in preoperative total bilirubin,the probability of developing postoperative bleeding complications would increase by 0.3％(OR=1.003,95％CI:1.001-1.004),and a preoperative total bilirubin＞264.4 μmol/L would have an impact on the postoperative bleeding.Conclusion The loop-forming location of draining tube and the drainage method are the independent risk factors for PTCD bleeding complications.The closer the loop-forming location to the tertiary branches is,the greater the risk of bleeding would be.The bleeding risk of internal-external joint drainage method is higher than that of external drainage method.The preoperative total bilirubin and preoperative platelet count are the independent risk factors for bleeding complications of PTCD.The preoperative total bilirubin level is positively correlated with bleeding risk,while the preoperative platelet count level is negatively correlated with the bleeding risk.(J Intervent Radiol,2024,33:500-506)

Objective:To analyze the influencing factors of urinary infection after radical cystectomy in elderly patients with bladder cancer.Methods:A retrospective study was conducted to collect the clinical data of 56 elderly patients with bladder cancer who developed urinary infection after radical cystectomy in the Second Affiliated Hospital of Xi′an Jiaotong University from March 2020 to February 2023. All patients were included in the case group. A total of 168 patients without urinary infection after radical cystectomy were collected at a ratio of 1∶3 into the control group. Baseline data and perioperative data were collected, and Logistic regression was used to analyze the influencing factors of urinary infection after radical cystectomy in elderly patients with bladder cancer.Results:Single factor analysis showed that the diabetes proportion, hydronephrosis proportion, nutrition risk proportion, inhalation anesthesia proportion, senile weakness proportion, stress hyperglycemia proportion, albumin (ALB) <35 g/L proportion and urine bag replacement time in the case group were significantly higher than those in the control group: 66.07% (37/56) vs. 44.64% (75/168), 69.64% (39/56) vs. 46.43% (78/168), 66.07% (37/56) vs. 41.67% (70/168), 73.21% (41/56) vs. 48.81% (82/168), 55.36% (31/56) vs. 35.12% (59/168), 41.07% (23/56) vs. 20.24% (34/168), 55.36% (31/56) vs. 36.90% (62/168), (7.52 ± 1.65) d vs. (6.62 ± 1.44) d, and there were statistical differences ( P<0.05). Logistic regression analysis showed that nutrition risk, inhalation anesthesia, stress hyperglycemia and senile weakness were independent factors of urinary infection after radical cystectomy in elderly patients with bladder cancer ( P<0.05). Conclusions:Urinary infection after radical cystectomy in the elderly patients with bladder cancer is closely related to nutritional risk, inhalation anesthesia, stress hyperglycemia and senile weakness. Attention to the above factors and symptomatic intervention are important for prevention and treatment of urinary infection.

Objective To observe the value of deep learning(DL)models for automatic classification of echocardiographic views.Methods Totally 100 patients after heart transplantation were retrospectively enrolled and divided into training set,validation set and test set at a ratio of 7∶2∶1.ResNet18,ResNet34,Swin Transformer and Swin Transformer V2 models were established based on 2D apical two chamber view,2D apical three chamber view,2D apical four chamber view,2D subcostal view,parasternal long-axis view of left ventricle,short-axis view of great arteries,short-axis view of apex of left ventricle,short-axis view of papillary muscle of left ventricle,short-axis view of mitral valve of left ventricle,also 3D and CDFI views of echocardiography.The accuracy,precision,recall,F1 score and confusion matrix were used to evaluate the performance of each model for automatically classifying echocardiographic views.The interactive interface was designed based on Qt Designer software and deployed on the desktop.Results The performance of models for automatically classifying echocardiographic views in test set were all good,with relatively poor performance for 2D short-axis view of left ventricle and superior performance for 3D and CDFI views.Swin Transformer V2 was the optimal model for automatically classifying echocardiographic views,with high accuracy,precision,recall and F1 score was 92.56％,89.01％,89.97％and 89.31％,respectively,which also had the highest diagonal value in confusion matrix and showed the best classification effect on various views in t-SNE figure.Conclusion DL model had good performance for automatically classifying echocardiographic views,especially Swin Transformer V2 model had the best performance.Using interactive classification interface could improve the interpretability of prediction results to some extent.

3D bioprinting technology is a rapidly developing technique that employs bioinks containing biological materials and living cells to construct biomedical products. However, 3D-printed tissues are static, while human tissues are in real-time dynamic states that can change in morphology and performance. To improve the compatibility between in vitro and in vivo environments, an in vitro tissue engineering technique that simulates this dynamic process is required. The concept of 4D printing, which combines "3D printing + time" provides a new approach to achieving this complex technique. 4D printing involves applying one or more smart materials that respond to stimuli, enabling them to change their shape, performance, and function under the corresponding stimulus to meet various needs. This article focuses on the latest research progress and potential application areas of 4D printing technology in the cardiovascular system, providing a theoretical and practical reference for the development of this technology.
Humans ; Tissue Engineering/methods* ; Bioprinting/methods* ; Printing, Three-Dimensional ; Cardiovascular System ; Tissue Scaffolds

Objective To investigate the clinical prognosis of the liver transplant recipients diagnosed with hepatocellular carcinoma (HCC) complicated with microvascular invasion (MVI). Methods Clinical data of 3 447 HCC recipients undergoing liver transplantation were extracted from Surveillance, Epidemiology, and End Results (SEER) database of American National Cancer Institute. According to the incidence of MVI, all recipients were divided into MVI (n=376) and non-MVI groups (n=3 071). The clinical prognosis of liver transplant recipients was statistically compared between two groups by analyzing the 1-, 3- and 5-year overall survival (OS) and liver cancer specific survival (LCSS). Relevant clinical data including age, gender, race, pathological staging, tumor size, lymph node metastasis, distant metastasis, tumor-node-metastasis (TNM) staging and MVI were recorded in two groups. The independent risk factors of clinical prognosis of HCC recipients undergoing liver transplantation were analyzed by multivariate Cox regression model. The nomogram for predicting the clinical prognosis of the recipients was delineated. The accuracy of the prediction model was evaluated by the consistency index. Results In the non-MVI group, the 1-, 3-, 5-year OS and LCSS were 93.5%, 82.1%, 75.3% and 98.3%, 93.8%, 90.7%, significantly higher than 88.8%, 72.1%, 68.4% and 95.3%, 83.1%, 80.4% in the MVI group (all P < 0.05). Multivariate regression analysis showed that pathological staging, tumor size, lymph node metastasis, distant metastasis, TNM staging and MVI were the independent risk factors of OS and LCSS in HCC recipients undergoing liver transplantation (all P < 0.05). The nomogram consistency index was calculated as 0.624 (0.602-0.648). Conclusions MVI is an independent risk factor of the clinical prognosis of HCC recipients undergoing liver transplantation, which is significantly correlated with poor prognosis of the recipients. The nomogram based on MVI can predict the clinical prognosis of these recipients.