Aim To construct and analyze the genotype of G protein-coupled receptor kinase 2(GRK2)conditional knockout mice in synoviocytes,and to provide an animal model for stud-ying the function of GRK2 in synoviocytes.Methods Grk2flox/+mice were bred to generate Grk2flox/flox mice,Grk2flox/flox mice were bred to Col1a1-iCre+mice,Grk2flox/+Col1a1-iCre+mice were bred to Grk2flox/flox mice.Grk2flox/flox Col1a1-iCre+mice were ob-tained as target mice.DNA was extracted and amplified by PCR to identify the genotype.Western blot was used to verify the effect of Grk2 knockout in synovium,liver and kidney tissues.HE staining was used to detect the effects of Grk2 conditional knockout in synovial cells on ankle synovium,liver and kidney tissues.Multiple immunofluorescence was used to detect GRK2 expression in synovial cells.Results The results of gene iden-tification showed that Grk2flox/flox Col1a1-iCre+mice had both Flox and Col1a1-iCre genotypes.Western blot results showed that GRK2 expression decreased in synovial tissues of Grk2flox/flox Col1a1-iCre+mice,but there was no significant change in the expression of GRK2 in liver and kidney tissues.HE staining showed that Grk2flox/flox Col1a1-iCre+mice had no significant pathological changes in the ankle synovium,liver and kidney.The results of multiple immunofluorescence showed that GRK2 expression in synovial cells of Grk2flox/flox Col1a1-iCre+mice de-creased.Conclusion Grk2 conditional knockout mice in syno-viocytes are successfully constructed and identified,which pro-vides an animal model for further study of the role of GRK2 in synovial-related diseases.

Aim To construct and analyze the genotype of G protein-coupled receptor kinase 2(GRK2)conditional knockout mice in synoviocytes,and to provide an animal model for stud-ying the function of GRK2 in synoviocytes.Methods Grk2flox/+mice were bred to generate Grk2flox/flox mice,Grk2flox/flox mice were bred to Col1a1-iCre+mice,Grk2flox/+Col1a1-iCre+mice were bred to Grk2flox/flox mice.Grk2flox/flox Col1a1-iCre+mice were ob-tained as target mice.DNA was extracted and amplified by PCR to identify the genotype.Western blot was used to verify the effect of Grk2 knockout in synovium,liver and kidney tissues.HE staining was used to detect the effects of Grk2 conditional knockout in synovial cells on ankle synovium,liver and kidney tissues.Multiple immunofluorescence was used to detect GRK2 expression in synovial cells.Results The results of gene iden-tification showed that Grk2flox/flox Col1a1-iCre+mice had both Flox and Col1a1-iCre genotypes.Western blot results showed that GRK2 expression decreased in synovial tissues of Grk2flox/flox Col1a1-iCre+mice,but there was no significant change in the expression of GRK2 in liver and kidney tissues.HE staining showed that Grk2flox/flox Col1a1-iCre+mice had no significant pathological changes in the ankle synovium,liver and kidney.The results of multiple immunofluorescence showed that GRK2 expression in synovial cells of Grk2flox/flox Col1a1-iCre+mice de-creased.Conclusion Grk2 conditional knockout mice in syno-viocytes are successfully constructed and identified,which pro-vides an animal model for further study of the role of GRK2 in synovial-related diseases.

Animals ; Antineoplastic Agents, Alkylating/pharmacology* ; Breast Neoplasms/drug therapy* ; Cell Line ; Cell Line, Tumor ; Cyclophosphamide/pharmacology* ; Ginkgo biloba/chemistry* ; Mammary Neoplasms, Animal/drug therapy* ; Mice ; Plant Extracts/administration & dosage*

Objective: To observe the characteristics and trends during the last 11 years of risk factors of young adults with first acute coronary syndrome (ACS). Methods: It was a cross-sectional study. We included young adults (18 to 44 years old) hospitalized for acute coronary syndrome in Beijing Anzhen Hospital for a first time from January 2007 to December 2017. Acute coronary syndromes include ST-elevation myocardial infarction (STEMI), non-ST-elevation myocardial infarction (NSTEMI), and unstable angina (UA). The general information, medical history and laboratory test were recorded. Risk factors of ACS were smoking, dyslipidemia, overweight/obesity, hypertension and diabetes. Results: Data from 7 106 patients were analyzed, mean age was (39.8±4.2) years old and 6 593(92.8%)were men, including 2 254 (31.7%) STEMI, 704 (9.9%) NSTEMI and 4 148 (58.4%) UA. Most patients were male (6 593(92.8%)). Dyslipidemia (85.8%(6 094/7 106)), overweight/obesity (82.3%(5 850/7 106)), and smoking (63.9%(4 545/7 106)) were most prevalent. 98.3% (6 885/7 106) patients had at least 1 risk factor. The prevalence of hypertension, diabetes and overweight/obesity increased from 2007 to 2017. Rates of hypertension increased from 37.1%(111/299) to 48.1%(498/1 035) (P_trend<0.01), diabetes from 12.0%(36/299) to 19.4%(201/1 035) (P_trend<0.01), overweight/obesity from 74.2%(222/299) to 83.9%(868/1 035) (P_trend<0.05), respectively. Conclusions: Dyslipidemia, overweight/obesity and smoking are most prevalent risk factors in young adults with a first ACS and most patients have at least 1 risk factor for ACS. Rates of hypertension, diabetes and overweight/obesity progressively increases over time in this patient cohort.

BACKGROUND: Micro-arc oxidation (MAO) treatment can improve the biological properties of titanium alloy implants. Previous studies mostly focused on the evaluation of titanium alloy plate, while the effects of the MAO-modified 3D titanium scaffold on the cell growth and differentiation were rarely reported. OBJECTIVE: To investigate the effect of MAO coating on the biological performance of cells seeded onto the 3D-printed porous titanium alloy scaffold. METHODS: Rat bone marrow mesenchymal stem cells were seeded onto the MAO-modified Ti6Al4V alloy scaffolds (experimental group) and unmodified scaffolds (control group). After 4 and 7 days of culture, cell/scaffold constructs were retrieved and processed for the assessment of cell morphology by using scanning electron microscopy, cytoskeletal staining analysis and cell viability assay were also evaluated. At 4, 7 and 11 days of culture, the levels of alkaline phosphatase and osteocalcin in the cell supernatant were detected. At 1, 4, 7 and 11 days of culture, the cell proliferation rate was measured using the MTT assay. RESULTS AND CONCLUSION: (1) At 4 and 7 days of culture, live/dead staining showed that the bone marrow mesenchymal stem cells grew well on the two kinds of scaffolds. The analysis of cytoskeleton staining showed that the cytoskeleton of the experimental group was stereo and polygonal, while the cells on the scaffold surface in the control group were flat and spindle-shaped, spreading along the macro structure of the scaffolds. Under the scanning electron microscopy, the cells in the experimental group arranged closely and spread in a good condition, with interconnected lamellipodia and filopodia that firmly adhered to the scaffold surface in an anchor-shaped structure; in the control group, less filopodia interconnected, less extracellular matrix, and flat and sheet-like cells were observed. (2) With the time increase, the levels of alkaline phosphatase and osteocalcin increased gradually in both groups. The alkaline phosphatase level in the experimental group was significantly higher than that in the control group at 7 and 11 days of culture (P < 0.05), while the osteocalcin level was higher in the experimental group than the control group at 11 days of culture (P < 0.05). (3) With the prolongation of culture time, the number of cells in the two groups increased gradually. The number of cells cultured in the experimental group was significantly higher than that in the control group at 7 and 11 days of culture (P < 0.05). To conclude, the MAO-modified titanium alloy scaffold is favorable for cell adhesion, proliferation and differentiation.

Adolescent ; Adult ; Aged ; Apoptosis ; Cell Cycle ; Cell Division ; Child ; Female ; Glioma ; chemistry ; pathology ; Humans ; Immunohistochemistry ; Inhibitor of Apoptosis Proteins ; Male ; Microtubule-Associated Proteins ; analysis ; Middle Aged ; Neoplasm Proteins ; Prognosis ; Proto-Oncogene Proteins c-bcl-2 ; analysis