OBJECTIVES: To compare the anti-inflammatory, antibacterial and analgesic effects of Yinqiao Powder (YQS) formulated granules and decoction. METHODS: We first evaluated the anti-inflammatory effects of the two dosage forms of YQS in a LPS-induced RAW 264.7 cell model using RT-qPCR and Western blotting. We further constructed zebrafish models of inflammation by copper sulfate exposure, caudal fin transection, or LPS and Poly (I:C) microinjection, and evaluated anti-inflammatory effects of YQS granules and decoction by examining neutrophil aggregation and HE staining findings. In a mouse model of acute lung injury (ALI) induced by intratracheal LPS instillation, the effects of YQS gavage at 10, 15, and 20 g/kg on lung pathologies were evaluated by calculating lung wet-dry weight ratio and using HE staining, ELISA and Western blotting. The microbroth dilution method was used to evaluate the antibacterial effect of YQS. Mouse pain models established by hot plate and intraperitoneal injection of glacial acetic acid were used to evaluate the analgesic effects of YQS at 10, 15, and 20 g/kg. RESULTS: Both YQS granules and decoction significantly reduced TNF-α, IL-6, and IL-1β expressions and p-STAT3 (Tyr 705) phosphorylation level in LPS-induced RAW 264.7 cells, and obviously inhibited neutrophil aggregation in the zebrafish models. In ALI mice, YQS granules and decoction effectively ameliorated lung injury, lowered lung wet-dry weight ratio, and reduced p-STAT3 (Tyr 705) expression and TNF-α and IL-6 levels. YQS produced obvious antibacterial effect at the doses of 15.63 and 31.25 mg/mL, and significantly reduced body torsion and increased pain threshold in the mouse pain models. CONCLUSIONS The two dosage forms of TQS have similar anti-inflammatory, antibacterial and analgesic effects with only differences in their inhibitory effect on TNF-α, IL-6 and IL-1β mRNA expressions in LPS-induced RAW 264.7 cells.
Animals ; Mice ; Drugs, Chinese Herbal/pharmacology* ; Anti-Inflammatory Agents/pharmacology* ; Analgesics/pharmacology* ; RAW 264.7 Cells ; Zebrafish ; Anti-Bacterial Agents/pharmacology* ; Powders ; Tumor Necrosis Factor-alpha/metabolism* ; Acute Lung Injury/drug therapy* ; Interleukin-6/metabolism* ; Lipopolysaccharides

BACKGROUND: The contraction behaviors of cardiomyocytes (CMs), especially contraction synchrony, are crucial factors reflecting their maturity and response to drugs. A wider field of view helps to observe more pronounced synchrony differences, but the accompanied greater computational load, requiring more computing power or longer computational time. METHODS: We proposed a method that directly correlates variations in optical field brightness with cardiac tissue contraction status (CVB method), based on principles from physics and photometry, for rapid video analysis in wide field of view to obtain contraction parameters, such as period and contraction propagation direction and speed. RESULTS: Through video analysis of human induced pluripotent stem cell (hiPSC)-derived CMs labeled with green fluorescent protein (GFP) cultured on aligned and random nanofiber scaffolds, the CVB method was demonstrated to obtain contraction parameters and quantify the direction and speed of contraction within regions of interest (ROIs) in wide field of view. The CVB method required less computation time compared to one of the contour tracking methods, the LucasKanade (LK) optical flow method, and provided better stability and accuracy in the results. CONCLUSION This method has a smaller computational load, is less affected by motion blur and out-of-focus conditions, and provides a potential tool for accurate and rapid analysis of cardiac tissue contraction synchrony in wide field of view without the need for more powerful hardware.

BACKGROUND: The contraction behaviors of cardiomyocytes (CMs), especially contraction synchrony, are crucial factors reflecting their maturity and response to drugs. A wider field of view helps to observe more pronounced synchrony differences, but the accompanied greater computational load, requiring more computing power or longer computational time. METHODS: We proposed a method that directly correlates variations in optical field brightness with cardiac tissue contraction status (CVB method), based on principles from physics and photometry, for rapid video analysis in wide field of view to obtain contraction parameters, such as period and contraction propagation direction and speed. RESULTS: Through video analysis of human induced pluripotent stem cell (hiPSC)-derived CMs labeled with green fluorescent protein (GFP) cultured on aligned and random nanofiber scaffolds, the CVB method was demonstrated to obtain contraction parameters and quantify the direction and speed of contraction within regions of interest (ROIs) in wide field of view. The CVB method required less computation time compared to one of the contour tracking methods, the LucasKanade (LK) optical flow method, and provided better stability and accuracy in the results. CONCLUSION This method has a smaller computational load, is less affected by motion blur and out-of-focus conditions, and provides a potential tool for accurate and rapid analysis of cardiac tissue contraction synchrony in wide field of view without the need for more powerful hardware.

BACKGROUND: The contraction behaviors of cardiomyocytes (CMs), especially contraction synchrony, are crucial factors reflecting their maturity and response to drugs. A wider field of view helps to observe more pronounced synchrony differences, but the accompanied greater computational load, requiring more computing power or longer computational time. METHODS: We proposed a method that directly correlates variations in optical field brightness with cardiac tissue contraction status (CVB method), based on principles from physics and photometry, for rapid video analysis in wide field of view to obtain contraction parameters, such as period and contraction propagation direction and speed. RESULTS: Through video analysis of human induced pluripotent stem cell (hiPSC)-derived CMs labeled with green fluorescent protein (GFP) cultured on aligned and random nanofiber scaffolds, the CVB method was demonstrated to obtain contraction parameters and quantify the direction and speed of contraction within regions of interest (ROIs) in wide field of view. The CVB method required less computation time compared to one of the contour tracking methods, the LucasKanade (LK) optical flow method, and provided better stability and accuracy in the results. CONCLUSION This method has a smaller computational load, is less affected by motion blur and out-of-focus conditions, and provides a potential tool for accurate and rapid analysis of cardiac tissue contraction synchrony in wide field of view without the need for more powerful hardware.

BACKGROUND:Hormonal osteonecrosis of the femoral head is strongly associated with aging,but the regulatory targets and mechanisms are still unclear.Through bioinformatics combined with machine learning analysis and experimental verification,the key genes of hormonal osteonecrosis of the femoral head mediated by cell senescence will be identified,which will provide new ideas for the prevention and treatment of hormonal osteonecrosis of the femoral head.OBJECTIVE:To screen and validate the senescence core genes of hormonal osteonecrosis of the femoral head using bioinformatics analysis to explore its mechanism of action.METHODS:The GSE123568 dataset was obtained from the GPL15207 platform of the GEO database,which contained the gene expression profiles of peripheral serum samples of 30 hormonal osteonecrosis of the femoral head patients and 10 healthy controls.Data on 279 cellular senescence-related genes were obtained from the CellAge database.Differential analysis and weighted correlation network analysis(WGCNA)were performed on hormonal osteonecrosis of the femoral head gene profiles,and both were intersected with senescence-related genes and then concatenated to obtain hormonal osteonecrosis of the femoral head senescence potential genes,and GO and KEGG analyses were performed.The machine learning method screened out the pivotal genes,constructed nomogram model,and performed consensus clustering and immune infiltration analysis.Finally,clinical femoral samples were collected for validation by qPCR and western blot assay.RESULTS AND CONCLUSION:(1)41 potential genes were obtained,which were mainly enriched in biological processes such as aging and oxidative stress response,as well as FoxO and tumor necrosis factor signaling pathways.(2)The pivotal genes catalase,connective tissue growth factor,forkhead box protein O3,insulin receptor substrate 2,and mitogen-activated protein kinase kinase 11 were obtained after machine learning identification,and the predictive ability of nomogram model was good.(3)The patients were classified into three groups,namely a,b and c,by the consensus clustering analysis.Catalase,forkhead box protein O3,insulin receptor substrate 2,and mitogen-activated protein kinase kinase 11 were differentially expressed among the three molecular subtypes(P＜0.05).Results of immune infiltration showed that the abundance of immune cells,such as activated CD4+T cells,activated CD8+T cells,and eosinophils,differed among the three molecular subclasses(P＜0.05).(4)The results of qPCR and western blot assay showed that the expression of catalase,connective tissue growth factor,forkhead box protein O3,and mitogen-activated protein kinase kinase 11 was lower in hormonal osteonecrosis of the femoral head group compared to the control group(P＜0.05),and the expression of insulin receptor substrate 2 was elevated(P＜0.05).(5)It is concluded that through in-depth analysis combined with bioinformatics and machine learning,and further experimental verification,five hormonal osteonecrosis of the femoral head age-related hub genes were finally identified.These genes are catalase,connective tissue growth factor,forkhead box o3,insulin receptor substrate 2,and serine/threonine kinase 11.These genes may provide potential molecular targets for the prevention and treatment of hormonal osteonecrosis of the femoral head in the future by regulating the cellular aging process.

Objective To investigate the incidence of intraoperative hypothermia(IH)and rewarming efficacy in elderly patients with different traditional Chinese medicine(TCM)constitutions undergoing general anesthesia.Methods A total of 500 elderly patients undergoing general anesthesia at the First Affiliated Hospital of Guangzhou University of Chinese Medicine from June 2022 to November 2022 were enrolled.The patients were divided into IH group and non-IH group depending on the occurrence of IH.Baseline data of the patients were collected,and univariate and multivariate logistic regression analyses were performed to identify risk factors for IH.A risk prediction model was constructed after value-assignment of influencing factors and its diagnostic efficiency was assessed by receiver operating characteristic(ROC)curve.External validation of the model was conducted in 50 elderly patients undergoing general anesthesia at the same period.From December 2022 to February 2023,80 elderly patients undergoing general anesthesia and complicated with IH were randomized into a control group(routine measures for keeping warm)and an observation group(keeping warm with forced-air-warming blanket)to compare the rewarming efficacy.Results(1)Among 500 elderly patients undergoing general anesthesia,180 cases developed IH,with an incidence of 36.0%.(2)Except for age,sex,type of surgery,American Society of Anesthesiologists(ASA)classification,type of anesthesia,and total fluid output volume,statistically significant differences were presented in body mass index(BMI),TCM constitution type,surgical complexity grading,preoperative core body temperature,operating room temperature,rewarming strategy,surgical duration,anesthesia duration,total fluid intake volume,intraoperative irrigation volume,and intraoperative blood transfusion between the two groups(P＜0.05 or P＜0.01).(3)Multivariate logistic regression analysis identified BMI,TCM constitution type,surgical complexity grading,preoperative core body temperature,operating room temperature,rewarming strategy,anesthesia duration,total fluid intake volume,and intraoperative irrigation volume as independent influencing factors for the occurrence of IH in elderly patients undergoing general anesthesia.ROC curve analysis demonstrated that the area under the curve(AUC)of the constructed risk prediction model was 0.816,with a sensitivity of 74.40%and a specificity of 77.50%.(4)In an external validation cohort of 50 elderly patients undergoing general anesthesia(19 IH cases versus 31 non-IH cases),the constructed risk prediction model exhibited a sensitivity of 73.68%and a specificity of 77.42%.(5)The observation group exhibited lower tympanic temperature upon post-anesthesia care unit(PACU)admission,shorter rewarming time and PACU stay duration,faster rewarming rate,and lower incidence of shivering than the control group,the differences being statistically significant(P＜0.05 or P＜0.01).Conclusion IH is common in elderly patients undergoing general anesthesia,particularly in those under the conditions of low BMI,qi/yang/blood deficiency constitutions,major surgery,low preoperative core body temperature,low operating room temperature,passive rewarming,prolonged anesthesia,high fluid intake volume,and high and intraoperative irrigation volume.The constructed risk prediction model demonstrates favorable diagnostic performance.Proactive use of forced-air-warming blankets effectively improves rewarming outcomes in elderly patients undergoing general anesthesia and complicated with IH.

BACKGROUND: The contraction behaviors of cardiomyocytes (CMs), especially contraction synchrony, are crucial factors reflecting their maturity and response to drugs. A wider field of view helps to observe more pronounced synchrony differences, but the accompanied greater computational load, requiring more computing power or longer computational time. METHODS: We proposed a method that directly correlates variations in optical field brightness with cardiac tissue contraction status (CVB method), based on principles from physics and photometry, for rapid video analysis in wide field of view to obtain contraction parameters, such as period and contraction propagation direction and speed. RESULTS: Through video analysis of human induced pluripotent stem cell (hiPSC)-derived CMs labeled with green fluorescent protein (GFP) cultured on aligned and random nanofiber scaffolds, the CVB method was demonstrated to obtain contraction parameters and quantify the direction and speed of contraction within regions of interest (ROIs) in wide field of view. The CVB method required less computation time compared to one of the contour tracking methods, the LucasKanade (LK) optical flow method, and provided better stability and accuracy in the results. CONCLUSION This method has a smaller computational load, is less affected by motion blur and out-of-focus conditions, and provides a potential tool for accurate and rapid analysis of cardiac tissue contraction synchrony in wide field of view without the need for more powerful hardware.

BACKGROUND: The contraction behaviors of cardiomyocytes (CMs), especially contraction synchrony, are crucial factors reflecting their maturity and response to drugs. A wider field of view helps to observe more pronounced synchrony differences, but the accompanied greater computational load, requiring more computing power or longer computational time. METHODS: We proposed a method that directly correlates variations in optical field brightness with cardiac tissue contraction status (CVB method), based on principles from physics and photometry, for rapid video analysis in wide field of view to obtain contraction parameters, such as period and contraction propagation direction and speed. RESULTS: Through video analysis of human induced pluripotent stem cell (hiPSC)-derived CMs labeled with green fluorescent protein (GFP) cultured on aligned and random nanofiber scaffolds, the CVB method was demonstrated to obtain contraction parameters and quantify the direction and speed of contraction within regions of interest (ROIs) in wide field of view. The CVB method required less computation time compared to one of the contour tracking methods, the LucasKanade (LK) optical flow method, and provided better stability and accuracy in the results. CONCLUSION This method has a smaller computational load, is less affected by motion blur and out-of-focus conditions, and provides a potential tool for accurate and rapid analysis of cardiac tissue contraction synchrony in wide field of view without the need for more powerful hardware.

BACKGROUND:Hormonal osteonecrosis of the femoral head is strongly associated with aging,but the regulatory targets and mechanisms are still unclear.Through bioinformatics combined with machine learning analysis and experimental verification,the key genes of hormonal osteonecrosis of the femoral head mediated by cell senescence will be identified,which will provide new ideas for the prevention and treatment of hormonal osteonecrosis of the femoral head.OBJECTIVE:To screen and validate the senescence core genes of hormonal osteonecrosis of the femoral head using bioinformatics analysis to explore its mechanism of action.METHODS:The GSE123568 dataset was obtained from the GPL15207 platform of the GEO database,which contained the gene expression profiles of peripheral serum samples of 30 hormonal osteonecrosis of the femoral head patients and 10 healthy controls.Data on 279 cellular senescence-related genes were obtained from the CellAge database.Differential analysis and weighted correlation network analysis(WGCNA)were performed on hormonal osteonecrosis of the femoral head gene profiles,and both were intersected with senescence-related genes and then concatenated to obtain hormonal osteonecrosis of the femoral head senescence potential genes,and GO and KEGG analyses were performed.The machine learning method screened out the pivotal genes,constructed nomogram model,and performed consensus clustering and immune infiltration analysis.Finally,clinical femoral samples were collected for validation by qPCR and western blot assay.RESULTS AND CONCLUSION:(1)41 potential genes were obtained,which were mainly enriched in biological processes such as aging and oxidative stress response,as well as FoxO and tumor necrosis factor signaling pathways.(2)The pivotal genes catalase,connective tissue growth factor,forkhead box protein O3,insulin receptor substrate 2,and mitogen-activated protein kinase kinase 11 were obtained after machine learning identification,and the predictive ability of nomogram model was good.(3)The patients were classified into three groups,namely a,b and c,by the consensus clustering analysis.Catalase,forkhead box protein O3,insulin receptor substrate 2,and mitogen-activated protein kinase kinase 11 were differentially expressed among the three molecular subtypes(P＜0.05).Results of immune infiltration showed that the abundance of immune cells,such as activated CD4+T cells,activated CD8+T cells,and eosinophils,differed among the three molecular subclasses(P＜0.05).(4)The results of qPCR and western blot assay showed that the expression of catalase,connective tissue growth factor,forkhead box protein O3,and mitogen-activated protein kinase kinase 11 was lower in hormonal osteonecrosis of the femoral head group compared to the control group(P＜0.05),and the expression of insulin receptor substrate 2 was elevated(P＜0.05).(5)It is concluded that through in-depth analysis combined with bioinformatics and machine learning,and further experimental verification,five hormonal osteonecrosis of the femoral head age-related hub genes were finally identified.These genes are catalase,connective tissue growth factor,forkhead box o3,insulin receptor substrate 2,and serine/threonine kinase 11.These genes may provide potential molecular targets for the prevention and treatment of hormonal osteonecrosis of the femoral head in the future by regulating the cellular aging process.

ObjectiveTo investigate the effects of Tongluo Juanbi granules on chondrocyte apoptosis and Toll-like receptor 4 （TLR4）/myeloid differentiation factor 88 （MyD88）/nuclear factor-κB （NF-κB） signaling pathway of rabbits with knee osteoarthritis （KOA） and study the mechanism of Tongluo Juanbi granules in the prevention and treatment of KOA. MethodThirty New Zealand rabbits were randomly assigned to the following five groups （n=6）： sham group， model group， low-dose and high-dose groups of Tongluo Juanbi granules （4.1 and 8.2 g·kg^-1·d^-1）， and celecoxib group （10.9 mg·kg^-1·d^-1）. The KOA model was established by destabilization of the medial meniscus （DMM） for six weeks. Six weeks after the modeling， the drug was given once a day for eight weeks. The pathological changes of cartilago articularis were observed by hematoxylin-eosin （HE） staining and Safranin O-Fast Green staining. Terminal deoxynucleotidyl transferase dUTP nick end labeling （TUNEL） staining was performed to detect chondrocyte apoptosis. Enzyme-linked immunosorbent assay （ELISA） was used to detect the contents of interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α） in synovial fluid. The mRNA and protein expression levels of genes related to the TLR4/MyD88/NF-κB signaling pathway were detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. ResultCompared with the sham group， the cartilago articularis of the model group significantly degenerated. Mankin's score was increased （P<0.01）， and the contents of IL-1β and TNF-α in synovial fluid were increased （P<0.01）. The number of apoptosis of chondrocytes was increased （P<0.01）. The mRNA and protein expressions of TLR4， MyD88， and NF-κB p65 in cartilage tissue were up-regulated （P<0.01）， while the mRNA and protein expressions of Bcl-2 were down-regulated （P<0.01）. Compared with the model group， chondrocyte degeneration in both low-dose and high-dose groups of Tongluo Juanbi granules was improved， and Mankin's score was decreased （P<0.01）. The contents of IL-1β and TNF-α were decreased （P<0.01）， and the number of apoptosis of chondrocytes was decreased （P<0.01）. The mRNA and protein expressions of TLR4， MyD88， and NF-κB p65 in cartilage tissue were down-regulated （P<0.01）， while the mRNA and protein expressions of Bcl-2 were up-regulated （P<0.01）. In addition， in the above observation indicators， the high-dose group of Tongluo Juanbi granules was significantly superior to the low-dose group of Tongluo Juanbi granules. ConclusionTongluo Juanbi granules could inhibit chondrocyte apoptosis in rabbits with KOA and improve cartilage degeneration， which may be related to inhibiting inflammatory responses mediated by TLR4/MyD88/NF-κB signaling pathway.