BACKGROUND:During bone remodeling,bone formation and bone resorption are spatially and temporally coordinated,involving intricate interactions between osteoclasts and osteoblasts.Isoginkgetin,a flavonoid found in Ginkgo biloba,has a wide range of anticancer activity and anti-reactive oxygen species activity;however,the effect of isoginkgetin on osteoclast differentiation is unknown.OBJECTIVE:To study the effect and mechanism of action of isoginkgetin on osteoclastogenesis.METHODS:In vitro studies were performed on mouse bone marrow-derived macrophages,and cell counting kit-8 cytotoxicity assay was used to detect the effect of isoginkgetin on cell viability of bone marrow-derived macrophages.Macrophage colony-stimulating factor and receptor activator of nuclear factor kappa-B ligand were used to induce the differentiation of bone marrow-derived macrophages to osteoclasts.Network pharmacology and molecular docking and molecular dynamics simulations were used to predict the processes and targets of the effects of isoginkgetin on the differentiation of osteoclasts.Tartrate-resistant acid phosphatase staining and F-actin staining were used to detect the effects of isoginkgetin on the differentiation and function of osteoclasts.Western blot and RT-PCR were used to detect the effects of isoginkgetin on the expression of genes and proteins related to osteoclast differentiation,reactive oxygen species,and PI3K/AKT pathways.Fluorescent probes were used to detect cellular and mitochondrial reactive oxygen species levels.Flow cytometry technology was used to detect reactive oxygen species levels in cells.RESULTS AND CONCLUSION:(1)Network pharmacology results showed that isoginkgetin affected osteoporosis mainly through the PI3K-AKT pathway and cellular response to drugs and hypoxia,and GSK3β,ESR1,MCL1 and CCNA2 were the key targets.(2)Cell counting kit-8 and tartrate-resistant acid phosphatase staining results showed that isoginkgetin at 8 μmol/L had the most significant inhibitory effect on osteoclastogenesis in vitro,and F-actin results showed that isoginkgetin inhibited osteoclast cytoskeletal actin ring formation in a concentration-dependent manner.(3)Molecular dynamics simulations showed that isoginkgetin bound well to osteoclastogenesis marker proteins(NFATc1,c-Fos,CTSK,and MMP9).Western blot and RT-PCR results indicated that isoginkgetin inhibited the expression of osteoclastogenesis marker proteins and genes(NFATc1,c-Fos,CTSK,and MMP9).(4)Western blot results showed that isoginkgetin inhibited the phosphorylation level of PI3K/AKT/GSK3β and suppressed osteoclastogenesis by activating the PI3K-AKT-GSK3β pathway.(5)The results of reactive oxygen species assay showed that isoginkgetin significantly reduced receptor activator of nuclear factor kappa-B ligand-induced cellular and mitochondrial reactive oxygen species production,and inhibited the differentiation of bone marrow-derived macrophages to osteoclasts.

BACKGROUND:During bone remodeling,bone formation and bone resorption are spatially and temporally coordinated,involving intricate interactions between osteoclasts and osteoblasts.Isoginkgetin,a flavonoid found in Ginkgo biloba,has a wide range of anticancer activity and anti-reactive oxygen species activity;however,the effect of isoginkgetin on osteoclast differentiation is unknown.OBJECTIVE:To study the effect and mechanism of action of isoginkgetin on osteoclastogenesis.METHODS:In vitro studies were performed on mouse bone marrow-derived macrophages,and cell counting kit-8 cytotoxicity assay was used to detect the effect of isoginkgetin on cell viability of bone marrow-derived macrophages.Macrophage colony-stimulating factor and receptor activator of nuclear factor kappa-B ligand were used to induce the differentiation of bone marrow-derived macrophages to osteoclasts.Network pharmacology and molecular docking and molecular dynamics simulations were used to predict the processes and targets of the effects of isoginkgetin on the differentiation of osteoclasts.Tartrate-resistant acid phosphatase staining and F-actin staining were used to detect the effects of isoginkgetin on the differentiation and function of osteoclasts.Western blot and RT-PCR were used to detect the effects of isoginkgetin on the expression of genes and proteins related to osteoclast differentiation,reactive oxygen species,and PI3K/AKT pathways.Fluorescent probes were used to detect cellular and mitochondrial reactive oxygen species levels.Flow cytometry technology was used to detect reactive oxygen species levels in cells.RESULTS AND CONCLUSION:(1)Network pharmacology results showed that isoginkgetin affected osteoporosis mainly through the PI3K-AKT pathway and cellular response to drugs and hypoxia,and GSK3β,ESR1,MCL1 and CCNA2 were the key targets.(2)Cell counting kit-8 and tartrate-resistant acid phosphatase staining results showed that isoginkgetin at 8 μmol/L had the most significant inhibitory effect on osteoclastogenesis in vitro,and F-actin results showed that isoginkgetin inhibited osteoclast cytoskeletal actin ring formation in a concentration-dependent manner.(3)Molecular dynamics simulations showed that isoginkgetin bound well to osteoclastogenesis marker proteins(NFATc1,c-Fos,CTSK,and MMP9).Western blot and RT-PCR results indicated that isoginkgetin inhibited the expression of osteoclastogenesis marker proteins and genes(NFATc1,c-Fos,CTSK,and MMP9).(4)Western blot results showed that isoginkgetin inhibited the phosphorylation level of PI3K/AKT/GSK3β and suppressed osteoclastogenesis by activating the PI3K-AKT-GSK3β pathway.(5)The results of reactive oxygen species assay showed that isoginkgetin significantly reduced receptor activator of nuclear factor kappa-B ligand-induced cellular and mitochondrial reactive oxygen species production,and inhibited the differentiation of bone marrow-derived macrophages to osteoclasts.

Objective To explore the rehabilitation effect of abdominal breathing training and the improvement status of cardiopulmonary function in elderly patients with chronic obstructive pulmonary disease (COPD) over 60 years old. Methods A total of 520 COPD patients over 60 years old admitted to the hospital were selected from June 2021 to March 2024 as the research subjects. The patients were divided into reference group (routine intervention, n=260) and abdominal rehabilitation group (abdominal breathing training on the basis of routine intervention, n=260) by adopting computer random function. Cardiopulmonary tolerance, respiratory muscle condition and pulmonary function markers were compared between the two groups before and after intervention. Results Compared with the reference group after intervention, the abdominal rehabilitation group had higher maximum oxygen uptake and minute ventilation volume, lower anaerobic threshold, and higher maximal expiratory pressure (MEP). The maximal inspiratory pressure (MIP), forced expiratory volume in one second (FEV1), forced expiratory volume (FVC), ratio of forced expiratory volume to forced vital capacity in one second (FEV1/FVC), the percentage of FEV1 to the predicted value (FEV1%pred) and 1 min maximum ventilation volume (MVV) (P<0.05) were all higher in the abdominal rehabilitation group than the reference group after intervention. Conclusion Abdominal breathing training for patients with COPD over 60 years old can effectively enhance cardiopulmonary tolerance, and improve respiratory muscle physiology and pulmonary function.

Objective To innovate and improve the feasibility of flexible ultrasonic cystoscopy (FUCS) in the diagnosis and treatment for urinary tract diseases in vivo by using female pigs as animal model.Methods Endobronchial ultrasonography was took as a substitute for FUCS and the operating skills and experience of FUCS in vivo by using female pigs as animal model were summarized,in order to testify the efficacy and safety of needle biopsy guided by FUCS.Results FUCS could simultaneously display the endoscopic images and ultrasonic images.Ninety ml was suitable in pig bladder for FUCS when the layers of bladder wall were clear.The mucosa,submucosa,muscularis and serosa layers were legible,which contributed to the needle positioning biopsy.Doppler ultrasound mode could distinguish the vessels,which could guide the puncture and avoid accidental injury of blood vessels and surrounding organs.Pathological results of positioning biopsy were consistent with FUCS evaluation for the three major layers of bladder wall.Conclusion Studies in vivo show that application of FUCS in the diagnosis and treatment for bladder disease is feasible,and further improvement of the FUCS equipment and clinical practice should be made.

OBJECTIVE: To investigate the effect of intranasal glucocorticoids treatment on the expression of c-fos and c-myc nasal polyps. METHOD: Immunohistochemistry method was used to determine c-fos and c-myc expression in nasal polyps from patients with topical steroids treatment for 10-12 weeks and untreated patients. RESULT: The rate of c-fos expressing cases was 15% and the rate of c-myc expressing cases was 20% in nasal polyps from topical steroid treated patients, but in untreated patients, the rate of c-fos expressing cases was 80% and the rate of c-myc expressing cases was 85%. There were significant differences between two groups (P < 0.01). The c-fos and c-myc expression was remarkably downregulated in nasal polyps from topical steroid treated patients compared to untreated patients. CONCLUSION The results show that glucocorticoids may induce cell apoptosis in nasal polyps by depressing the c-fos and c-myc expression.
Adult ; Budesonide ; administration & dosage ; therapeutic use ; Case-Control Studies ; Female ; Glucocorticoids ; therapeutic use ; Humans ; Male ; Nasal Polyps ; drug therapy ; metabolism ; Proto-Oncogene Proteins c-fos ; metabolism ; Proto-Oncogene Proteins c-myc ; metabolism

Objective To establish an orthotopic animal model of human prostate carcinoma (PC),and to provide a useful tool for the research of PC. Methods A total of 30 male BALB/c nude mice were included.First,2?10 6 human PC-3 cells were injected subcutaneously in 10 nude mice.Then 8 weeks later,the tumor on mice backs were took out and smashed into pieces.The tissue was implanted on the ventral lateral lobe of prostate gland by surgical orthotopic implantation in the other 20 nude mice.Twelve weeks later, the mice were autopsied,and histopathological examination of orthotopic tumors and relevant organs was done to identify and quantitate micrometastasis. Results Of the 20 mice,18 (90%) had tumor growth.The tumors were greater than 1.5 cm in diameter in 17 mice.Dilatation of bladder and hydronephrosis because of obstruction occurred in 12 mice.A high frequency of lymph node and organ metastasis was noted upon histological examination (10 with retroperitoneal para-aortic lymph node metastasis,4 with lung metastasis,1 with liver metastasis).But no bone metastasis was found. Pathological sections showed that most parts of the prostate were occupied by tumor cells. The nuclei looked polymorphic and hyperchromatic.Abnormal nucleus division could be seen.The cortices and medullae of metastatic lymph nodes were occupied by tumor cells. Conclusions The model set up by surgical orthotopic implantation can show the biological features of prostate carcinoma cells well.It expresses the metastatic potential of PC cell line significantly and can be considered an ideal heterotransplant animal model.