Objective This study aimed to compare the osteogenic performance differences of titanium surface coat-ings modified by dopamine or silanized graphene oxide,and to provide a more suitable modification scheme for ti-tanium surface graphene oxide coatings.Methods Tita-nium was subjected to alkali-heat treatment and then modified with dopamine and silanization,respectively,followed by coating with graphene oxide.Control and ex-perimental groups were designed as follows:pure titanium(Ti)group;titanium after alkali-heat treatment(Ti-NaOH)group;titanium after alkali-heat treatment and silanization modification(Ti-APTES)group;titanium after alkali-heat treatment and dopamine modification(Ti-DOPA)group;titanium with silanization-modified surface decorated with gra-phene oxide(Ti-APTES/GO)group;titanium with dopamine-modified surface decorated with graphene oxide(Ti-DOPA/GO)group.The physical and chemical properties of the material surfaces were analyzed using scanning electron micros-copy(SEM),contact angle goniometer,X-ray photoelectron spectroscopy(XPS),and Raman spectrometer.The prolifera-tion and adhesion morphology of mouse embryonic osteoblast precursor cells MC3T3-E1 on the material surfaces were observed by cell viability detection and immunofluorescence staining followed by laser confocal microscopy.The effects on the osteogenic differentiation of MC3T3-E1 cells were studied by alkaline phosphatase(ALP)staining,alizarin red staining and quantification,and real-time quantitative polymerase chain reaction.Results After modification with gra-phene oxide coating,a thin-film-like structure was observed on the surface under SEM.The hydrophilicity of all experi-mental groups was improved,among which the Ti-DOPA/GO group had the best hydrophilicity.XPS and Raman spec-troscopy analysis showed that the modified materials exhibited typical D and G peaks,and XPS revealed the presence of a large number of oxygen-containing functional groups on the surface.CCK8 assay showed that all groups of materials had no cytotoxicity,and the proliferation level of the Ti-APTES/GO group was higher than that of the Ti-DOPA/GO group.Under the laser confocal microscope,the cells in the Ti-DOPA/GO and Ti-APTES/GO groups spread more fully.The Ti-DOPA/GO and Ti-APTES/GO groups had the deepest ALP staining,and the Ti-APTES/GO group had the most alizarin red-stained mineralized nodules and the highest quantitative result of alizarin red staining.In the Ti-DOPA/GO and Ti-APTES/GO groups,the expression of the early osteogenic-related gene RUNX2 reached a relatively high level,while in the expression of the late osteogenic-related genes OPN and OCN,the Ti-APTES/GO group performed better than the Ti-DOPA/GO group.Conclusion Ti-APTES/GO significantly outperformed Ti-DOPA/GO in promoting the adhesion,proliferation,and in vitro osteogenic differentiation of MC3T3-E1 cells.

OBJECTIVES: This study aimed to compare the osteogenic performance differences of titanium surface coatings modified by dopamine or silanized graphene oxide, and to provide a more suitable modification scheme for titanium surface graphene oxide coatings. METHODS: Titanium was subjected to alkali-heat treatment and then modified with dopamine and silanization, respectively, followed by coating with graphene oxide. Control and experimental groups were designed as follows: pure titanium (Ti) group; titanium after alkali-heat treatment (Ti-NaOH) group; titanium after alkali-heat treatment and silanization modification (Ti-APTES) group; titanium after alkali-heat treatment and dopamine modification (Ti-DOPA) group; titanium with silanization-modified surface decorated with graphene oxide (Ti-APTES/GO) group; titanium with dopamine-modified surface decorated with graphene oxide (Ti-DOPA/GO) group. The physical and chemical properties of the material surfaces were analyzed using scanning electron microscopy (SEM), contact angle goniometer, X-ray photoelectron spectroscopy (XPS), and Raman spectrometer. The proliferation and adhesion morphology of mouse embryonic osteoblast precursor cells MC3T3-E1 on the material surfaces were observed by cell viability detection and immunofluorescence staining followed by laser confocal microscopy. The effects on the osteogenic differentiation of MC3T3-E1 cells were studied by alkaline phosphatase (ALP) staining, alizarin red staining and quantification, and real-time quantitative polymerase chain reaction. RESULTS: After modification with graphene oxide coating, a thin-film-like structure was observed on the surface under SEM. The hydrophilicity of all experimental groups was improved, among which the Ti-DOPA/GO group had the best hydrophilicity. XPS and Raman spectroscopy analysis showed that the modified materials exhibited typical D and G peaks, and XPS revealed the presence of a large number of oxygen-containing functional groups on the surface. CCK8 assay showed that all groups of materials had no cytotoxicity, and the proliferation level of the Ti-APTES/GO group was higher than that of the Ti-DOPA/GO group. Under the laser confocal microscope, the cells in the Ti-DOPA/GO and Ti-APTES/GO groups spread more fully. The Ti-DOPA/GO and Ti-APTES/GO groups had the deepest ALP staining, and the Ti-APTES/GO group had the most alizarin red-stained mineralized nodules and the highest quantitative result of alizarin red staining. In the Ti-DOPA/GO and Ti-APTES/GO groups, the expression of the early osteogenic-related gene RUNX2 reached a relatively high level, while in the expression of the late osteogenic-related genes OPN and OCN, the Ti-APTES/GO group performed better than the Ti-DOPA/GO group. CONCLUSIONS Ti-APTES/GO significantly outperformed Ti-DOPA/GO in promoting the adhesion, proliferation, and in vitro osteogenic differentiation of MC3T3-E1 cells.
Titanium/chemistry* ; Graphite/chemistry* ; Dopamine/chemistry* ; Animals ; Mice ; Osteogenesis ; Osteoblasts/cytology* ; Surface Properties ; Cell Proliferation ; Silanes/chemistry* ; Cell Adhesion ; Coated Materials, Biocompatible/chemistry* ; Cell Differentiation ; Alkaline Phosphatase/metabolism* ; Microscopy, Electron, Scanning

Flowering and bolting are important agronomic traits in cruciferous crops such as Brassica juncea. Timely flowering can ensure the crop organ yield and quality, as well as seed propagation. The WRKY family plays an important role in regulating plant bolting and flowering, while the function and mechanism of WRKY12 in B. juncea remain unknown. To explore its function and mechanism in bolting and flowering of B. juncea, we cloned and characterized the BjuWRKY12 gene in B. juncea and found that its expression levels were significantly higher in flowers and inflorescences than in leaves. BjuWRKY12 belonged to the Ⅱc subfamily of the WRKY family, and subcellular localization indicated that the protein was located in the nucleus. Ectopic overexpression of BjuWRKY12 in transgenic lines promoted bolting and flowering, leading to significant increases in the expression levels of flowering integrators SOC1 and FUL. Furthermore, yeast one-hybrid and dual luciferase reporter system assays confirmed that BjuWRKY12 directly bound to the promoters of BjuSOC1 and BjuFUL, undergoing protein-DNA interactions. This discovery gives new insights into the regulation network and molecular mechanisms of BjuWRKY12, laying a theoretical foundation for the breeding of high-yield and high-quality varieties of B. juncea.
Mustard Plant/metabolism* ; Flowers/growth & development* ; Plant Proteins/physiology* ; Promoter Regions, Genetic/genetics* ; Gene Expression Regulation, Plant ; Plants, Genetically Modified/genetics* ; Transcription Factors/metabolism* ; MADS Domain Proteins/metabolism*

OBJECTIVE To investigate and analyze the protective effect of Huayu Jiedu Decoction(HYJD)on the inflammatory injury of cardiomyocytes induced by bacterial lipopolysaccharide(LPS),and its molecular mecha-nism of inhibitory effect on inflammatory response.METHODS H9c2 cells were cultured in vitro and divided into:the blank control group(Control group),the model control group(LPS group),the drug treatment group(HYJD group)and the combined treatment group(LPS+HYJD group).H9c2 cells were treated with different concentrations of HYJD(2.5,5,10,20 and 40 mg/ml)for 24 h,and the activity of H9c2 cells was detected by MTT assay.Additionally,H9c2 cells were treated with LPS-induced myocardial inflammatory injury cell model after 24 h of HYJD intervention at each concentration gradients to detect the cell proliferation changes,as well as to detect the levels of apoptosis of cardiomyocytes and the levels of interleukin(IL)-1β,IL-6,IL-8,IL-10 and tumor necrosis factor(TNF)-α in the culture supernatant of experimental groups,the changes in the protein ex-pression of NO production and the expression changes of iNOS and TLR4/NF-κB signaling pathway protein,and Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect mRNA expression of IL-7R,P38(MAPK)and CXCR2.RESULTS Compared with the Control group,low-concentration HYJD had no significant effect on H9c2 cell viability and did not induce cytotoxic effect,and HYJD increased the survival rate of H9c2 cells in the LPS-induced myocardial inflammatory injury model,and effectively reversed the inhibitory effect of the pro-liferative activity of H9c2 cells induced by LPS.Compared with the control group,the difference in apoptosis level of H9c2 cells in the HYJD monotherapy group was not statistically significant,while the levels of inflammatory apoptosis of H9c2 cells induced by LPS was elevated(P＜0.05).Compared with the LPS group,HYJD inhibited the levels of inflammatory apoptosis in H9c2 cells induced by LPS(P＜0.05),reduced the production of pro-in-flammatory cytokines such as IL-1β,IL-6,IL-8 and TNF-α in the supernatant of the LPS-induced myocardial in-flammatory injury H9c2 cell culture,and upregulated the anti-inflammatory cytokine IL-10.Additionally,com-pared with the Control group,the LPS group showed an increased level of NO release(P＜0.05),while the difference in NO release in the low-concentration(5 mg/ml)HYJD was not statistically significant.Compared with the LPS group,the NO release levels in each HYJD intervention group showed a concentration-dependent de-crease(all P＜0.05).Furthermore,compared with the control group,whereas the expression levels of iNOS and TLR4/NF-κB signaling pathway proteins in the LPS-induced H9c2 cells were both elevated(P＜0.05).CONCLUSION HYJD exhibits protective effects against LPS-induced septic myocardial injury and can exert an in-hibitory effect on inflammatory response,and the molecular mechanisms may be related to the inhibition of the ac-tivation of the TLR4/NF-κB signaling pathway and the down-regulation of the expression of inflammatory genes,etc.,and it may have a good biological activity in the prevention and treatment of septic myocardial injury.

OBJECTIVE To investigate and analyze the protective effect of Huayu Jiedu Decoction(HYJD)on the inflammatory injury of cardiomyocytes induced by bacterial lipopolysaccharide(LPS),and its molecular mecha-nism of inhibitory effect on inflammatory response.METHODS H9c2 cells were cultured in vitro and divided into:the blank control group(Control group),the model control group(LPS group),the drug treatment group(HYJD group)and the combined treatment group(LPS+HYJD group).H9c2 cells were treated with different concentrations of HYJD(2.5,5,10,20 and 40 mg/ml)for 24 h,and the activity of H9c2 cells was detected by MTT assay.Additionally,H9c2 cells were treated with LPS-induced myocardial inflammatory injury cell model after 24 h of HYJD intervention at each concentration gradients to detect the cell proliferation changes,as well as to detect the levels of apoptosis of cardiomyocytes and the levels of interleukin(IL)-1β,IL-6,IL-8,IL-10 and tumor necrosis factor(TNF)-α in the culture supernatant of experimental groups,the changes in the protein ex-pression of NO production and the expression changes of iNOS and TLR4/NF-κB signaling pathway protein,and Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect mRNA expression of IL-7R,P38(MAPK)and CXCR2.RESULTS Compared with the Control group,low-concentration HYJD had no significant effect on H9c2 cell viability and did not induce cytotoxic effect,and HYJD increased the survival rate of H9c2 cells in the LPS-induced myocardial inflammatory injury model,and effectively reversed the inhibitory effect of the pro-liferative activity of H9c2 cells induced by LPS.Compared with the control group,the difference in apoptosis level of H9c2 cells in the HYJD monotherapy group was not statistically significant,while the levels of inflammatory apoptosis of H9c2 cells induced by LPS was elevated(P＜0.05).Compared with the LPS group,HYJD inhibited the levels of inflammatory apoptosis in H9c2 cells induced by LPS(P＜0.05),reduced the production of pro-in-flammatory cytokines such as IL-1β,IL-6,IL-8 and TNF-α in the supernatant of the LPS-induced myocardial in-flammatory injury H9c2 cell culture,and upregulated the anti-inflammatory cytokine IL-10.Additionally,com-pared with the Control group,the LPS group showed an increased level of NO release(P＜0.05),while the difference in NO release in the low-concentration(5 mg/ml)HYJD was not statistically significant.Compared with the LPS group,the NO release levels in each HYJD intervention group showed a concentration-dependent de-crease(all P＜0.05).Furthermore,compared with the control group,whereas the expression levels of iNOS and TLR4/NF-κB signaling pathway proteins in the LPS-induced H9c2 cells were both elevated(P＜0.05).CONCLUSION HYJD exhibits protective effects against LPS-induced septic myocardial injury and can exert an in-hibitory effect on inflammatory response,and the molecular mechanisms may be related to the inhibition of the ac-tivation of the TLR4/NF-κB signaling pathway and the down-regulation of the expression of inflammatory genes,etc.,and it may have a good biological activity in the prevention and treatment of septic myocardial injury.

Objective To explore the clinical value of models based on clinical features and enhanced MRI radiomics for predicting the occurrence of post-acute pancreatitis diabetes mellitus(PPDM-A).Methods A retrospective selection of 161 acute pancreatitis(AP)patients was conducted,comprising 99 in the non-PPDM-A group and 62 in the PPDM-A group.They were randomly divided into training set and test set in a ratio of 7∶3.Region of interest(ROI)were delineated and radiomics features were extracted on the late arterial phase MRI images.Optimal radiomics features were selected by maximum relevance and minimum redundancy(mRMR)and least absolute shrinkage and selection operator(LASSO).Support vector machine(SVM)was used to develop three predictive models.The efficacy of the models in predicting PPDM-A was evaluated,the receiver operating characteristic(ROC)curve was drawn,and the DeLong test was employed to assess the difference in predictive capability among the models.Results In the training set,the area under the curve(AUC)of the clinical model,radiomics model,and combined model were 0.702,0.810 and 0.901,respectively,and in the test set were 0.678,0.797 and 0.830,respectively.The DeLong test revealed a statistically significant difference in the predictive capability of the combined model compared to the clinical model both in the training and test sets(training set:P＜0.001;test set:P=0.019).Conclusion The combined model based on clinical features and enhanced MRI radiomics features demonstrates good predictive effi-cacy and can provide valuable insights for clinical interventions aimed at preventing PPDM-A.

Objective To explore the value of dual-phase enhanced CT radiomics in predicting post-acute pancreatitis diabetes mellitus(PPDM-A).Methods A total of 145 patients with acute pancreatitis(AP)were retrospectively collected,including 62 patients in PPDM-A group and 83 patients in non-PPDM-A group.The patients were randomly divided into training set and test set at a ratio of 7︰3,the pancreatic parenchyma in arterial phase and venous phase was delineated and the radiomics features were extracted.Vari-ance threshold method,univariate selection method and least absolute shrinkage and selection operator(LASSO)were used to screen radiomics features.The prediction performance of the model was evaluated by the area under the curve(AUC).The DeLong test was used to compare the prediction efficiency between the models,and the calibration curve and decision curve were used to evaluate the prediction efficiency of the model.Results The AUC of arterial phase model,venous phase model,combined arterial venous phase model,clinical model and radiomics combined clinical model in the training set were 0.845,0.792,0.829,0.656 and 0.862,respec-tively.The DeLong test results showed that only the difference between the radiomics combined clinical model and the clinical model in the training set and the test set was statistically significant(P<0.05).The decision curve showed that the radiomics combined clinical model had high clinical practicability in a certain range,and the calibration curve showed that the radiomics combined clinical model had the best fitting degree with the actual observation value.Conclusion Based on the dual-phase enhanced CT radiomics combined clinical model,PPDM-A can be predicted more accurately,and it can provide a certain reference value for the clinical development of per-sonalized treatment programs.

Objective:To investigate the diagnostic value of dynamic contrast enhanced MRI (DCE-MRI) quantitative parameters for preoperative staging of gastric cancer and its relationship with prognostic factors.Methods:The clinical data of 98 patients with gastric cancer from March 2021 to March 2022 in Guangyuan First People′s Hospital were retrospectively analyzed. All patients underwent DCE-MRI examination, MRI features were observed, and the DCE-MRI quantitative parameters were recorded, including the transport constant (K trans), volume fraction (V e) and rate constant (K ep). The expression levels of human epidermal growth factor receptor 2 (HER2) and epidermal growth factor receptor (EGFR) in gastric cancer tissue were detected by immunohistochemistry methods. The correlation between DCE-MRI quantitative parameters and T stage, HER2, EGFR of gastric cancer was analyzed by Spearman method; the receiver operating characteristic (ROC) curve was used to evaluate the diagnosis value of DCE-MRI quantitative parameters in T staging of gastric cancer. Results:Among 98 patients with gastric cancer, T 1 to T 2 staging was in 50 cases, T 3 to T 4 staging was in 48 cases; HER2 positive expression in gastric cancer tissue was in 35 cases, negative expression was in 63 cases; EGFR positive expression in gastric cancer tissue was in 43 cases, negative expression was in 55 cases. The K trans and V e in patients with T 3 to T 4 staging were significantly higher than those in patients with T 1 to T 2 staging: (0.25 ± 0.04) min -1 vs. (0.19 ± 0.03) min -1 and 0.45 ± 0.11 vs. 0.39 ± 0.09, and there were statistical differences ( P<0.01); there was no statistical difference in K ep between the two ( P>0.05). The K trans and V e in patients with HER2 positive expression were significantly higher than those in patients with HER2 negative expression: (0.27 ± 0.06) min -1 vs. (0.19 ± 0.03) min -1 and 0.49 ± 0.13 vs. 0.38 ± 0.08, and there were statistical differences ( P<0.01); there was no statistical difference in K ep between the two ( P>0.05). The K trans and V e in patients with EGFR positive expression were significantly higher than those in patients with EGFR negative expression: (0.28 ± 0.07) min -1 vs. (0.17 ± 0.04) min -1 and 0.50 ± 0.14 vs. 0.36 ± 0.08, and there were statistical differences ( P<0.01); there was no statistical difference in K ep between the two ( P>0.05). Spearman analysis result showed that the K trans was positively correlated with gastric cancer T stage, and the expression of HER2, EGFR in gastric cancer tissue ( r = 0.539, 0.612 and 0.640; P<0.01), the V e was positively correlated with gastric cancer T stage, and the expression of HER2, EGFR in gastric cancer tissue ( r = 0.462, 0.551 and 0.583; P<0.01), while there was no correlated between K ep and gastric cancer T stage and the expression of HER2, EGFR in gastric cancer tissue ( P>0.05). ROC curve analysis result showed that the area under the curve of K trans combined with V e in diagnosis the T 3 to T 4 staging of gastric cancer was 0.929, with a specificity of 81.25% and a specificity of 92.00%. Conclusions:The DCE-MRI quantitative parameters K trans and V e have certain value in the diagnosis of gastric cancer T staging, and they are closely related to the expression of prognostic factors HER2 and EGFR.

Periarticular fracture of the shoulder is a common type of fractures in the elderly. Postoperative adverse events such as internal fixation failure, humeral head ischemic necrosis and upper limb dysfunction occur frequently, which seriously endangers the exercise and health of the elderly. Compared with the fracture with normal bone mass, the osteoporotic periarticular fracture of the shoulder is complicated with slow healing and poor rehabilitation, so the clinical management becomes more difficult. At present, there is no targeted guideline or consensus for this type of fracture in China. In such context, experts from Youth Osteoporosis Group of Chinese Orthopedic Association, Orthopedic Expert Committee of Geriatrics Branch of Chinese Association of Gerontology and Geriatrics, Osteoporosis Group of Youth Committee of Chinese Association of Orthopedic Surgeons and Osteoporosis Committee of Shanghai Association of Chinese Integrative Medicine developed the Chinese expert consensus on the diagnosis and treatment of osteoporotic periarticular fracture of the shoulder in the elderly ( version 2023). Nine recommendations were put forward from the aspects of diagnosis, treatment strategies and rehabilitation of osteoporotic periarticular fracture of the shoulder, hoping to promote the standardized, systematic and personalized diagnosis and treatment concept and improve functional outcomes and quality of life in elderly patients with osteoporotic periarticular fracture of the shoulder.

Background/Aims: This study aimed to investigate the biological function and regulatory mechanism of TCN1 in colorectal cancer (CRC). Methods: We studied the biological function of TCN1 by performing gain-of-function and loss-offunction analyses in HCT116 cell lines; examined the effects of TCN1 on the proliferation, apoptosis, and invasion of CRC cells; and determined potential molecular mechanisms using HCT116 and SW480 CRC lines and mouse xenotransplantation models. Tumor xenograft and colonization assays were performed to detect the tumorigenicity and metastatic foci of cells in vivo. Results: TCN1 knockdown attenuated CRC cell proliferation and invasion and promoted cell apoptosis. Overexpression of TCN1 yielded the opposite effects. In addition, TCN1-knockdown HCT116 cells failed to form metastatic foci in the peritoneum after intravenous injection. Molecular mechanism analyses showed that TCN1 interacted with integrin subunit β4 (ITGB4) to positively regulate the expression of ITGB4. TCN1 knockdown promoted the degradation of ITGB4 and increased the instability of ITGB4 and filamin A. Downregulation of ITGB4 at the protein level resulted in the disassociation of the ITGB4/plectin complex, leading to cytoskeletal damage. Conclusions TCN1 might play an oncogenic role in CRC by regulating the ITGB4 signaling pathway.