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Objective To determine the role of AAMP in osteosarcoma cells and explore the mechanism of invasion and metastasis of osteosarcoma cells regulated by AAMP through the YAP signaling pathway. Methods Public sequencing data analysis was used to explore the correlation between AAMP and osteosarcoma. q-PCR, Western blot, and immunohistochemistry were used to detect the expression levels of osteosarcoma cell-related molecules. CCK-8 was used to detect cell proliferation ability. Transwell and wound healing assays were used to detect the invasive and metastatic abilities of osteosarcoma cells. Immunofluorescence was used to detect the cell localization and expression levels of related molecules. Results High expression of AAMP was negatively correlated with the prognosis of patients with osteosarcoma (P<0.05), and the expression of AAMP in patients with metastatic osteosarcoma increased (P<0.05). Compared with the control group, the AAMP interference group showed significantly decreased migratory, invasive, and EMT activities (P<0.05). The expression of p-CFL1 reduced after the knockdown of AAMP, and the cell plate pseudopods decreased significantly (P<0.05). A positive correlation was found between the expression levels of AAMP and YAP in osteosarcoma cells (P<0.05). Interfering with YAP expression can affect the migration and invasion of osteosarcoma cells. Conclusion AAMP promotes osteosarcoma cell metastasis by regulating the YAP signaling pathway, suggesting that AAMP may be a key molecule in promoting invasion and metastasis of osteosarcoma.
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The present study was conducted to compare the performance of broiler reared under two different light sources and three different light colours. For this purpose, 120, two-week-old IBL-80 (Indian Broiler Ludhiana-80) broiler chicks were randomly distributed in four different treatment groups viz. TLEDB-G (first 2 wks, blue LED then switched to green LED for the next 2 wks), TLEDG-B (first 2 wks, green LED then switch to blue LED for the next 2 wks), TLEDW (White LED) and TCFL (CFL light; Control) with 3 replications and 10 birds in each experimental unit was applied. The effects of different lights on performance (BW, BWG and FCR), carcass traits and its economic impact on broiler chickens were investigated in the present study. The results show that performance and carcass traits of broiler birds of blue-green and green blue LED light group was at par to that of CFL group whereas benefit cost ratio of birds of TLEDB-G (1.13) was found highest among different treatment groups. Therefore, use of a combination of monochromatic Blue-Green or Green-Blue LED light could be a better alternative source of light than CFL light in terms of birds’ performance, economics and energy saving
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OBJECTIVE: To establish a cell model stably expressing mouse organic anion transporter1( OAT1) in MDCK cells, for the purpose of screening potent OAT1 inhibitors in vitro. METHODS: Recombinant plasmid pcDNA3.1(+) -OAT1 was constructed and transfected into MDCK cells using Lipofectamine™ 2000 reagent. After the process of G418 screening, cells were collected for further validation. Cells were harvested, and the quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) was carried out to test the OAT1 mRNA expression in MDCK-OAT1 cells. The function of the stably transfected cells were validated by the uptake activity of (6-Carboxyfluorescein, 6-CFL),a substrate of OAT1. The inhibitors of OAT1 were selected according to their inhibition activity towards the uptake of 6-CFL into the OAT1-over expressing cells in comparison with the typical inhibitor of OAT1,probenecid. RESULTS: The pcDNA3.1(+) -OAT1 was well conducted. The mRNA expression of OAT1 was significantly higher than that in mock cells; MDCK-OAT1 cells had a significantly high mRNA expression comparing with the mock cells, being 4 862 fold of that in mock cells. The uptake-ability of 6-CFL in MDCK-OAT1 and MDCK-mock cells was obviously different, with a 14.9 fold increase in comparison with mock cells. In the presence of probenecid and several monomers from Chinese herbs, fluorescence values in cell lysates were reduced to varying degrees, and results showed that rhein, luteolin, chrysin and quercetin could significantly inhibited the 6-CFL uptake mediated by hOAT1,with a reduction of more than 80% of the control. CONCLUSION: The aim to establish a cell model which could stably express OAT1 is achieved. Further study could be done using this cell model, for the screening of potential inhibitors of OAT1 from monomers of Chinese herbs,and then could be used as a tool in the research of herb-drug interaction.
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Purpose To clarify the morphological parameter and describe the distance from the insertion of the lateral ankle ligaments to the adjacent bony landmarks through precisely anatomical explore of human cadaveric ankles,so as to provide anatomical evidences for the reconstruction of lateral ankle ligaments.Methods Nineteen ankle specimens were dissected to isolate the lateral ankle ligaments and measure the morphological parameters such as length,width,thickness and the distance from the insertion of the lateral ankle ligaments to the adjacent bony landmarks.Results The average length of anterior talofibular ligaments (ATFL) was 23.1 ± 2.98 mm,among which 8 were single-banded(42.1％)and 11 were double-banded(57.9％).The average distance from the fibular origination of ATFL to the anterior tubercle of fibula(AA)was 17.1 ± 3.00 mm,to the fibular obscure tubercle(AO)was 5.1 ± 1.69 mm,to the tip of the fibula(AT)was 14.1 ± 2.86 mm.The distances from the talus insertion of ATFL to the superior and inferior talus articular surface were 11.4 ± 2.25 mm and 18.4 ± 2.30 mm respectively,to the anterior lateral talus chondral surface was 4.8 ± 1.42 mm.The average length of calcaneofibular ligament(CFL)was 31.4 ± 3.55 mm.The average distance of the fibular origination from ATFL to CFL was 6.4 ± 2.55 mm.The average angle between ATFL and CFL was 116.6 ± 12.69°.The distance from the calcaneus insertion of CFL to the peroneal tubercle(CP)was 15.4 ± 2.86 mm,to the posterior superior border of calcaneus(CC)was 13.9 ± 2.46 mm,to the subtalar joint surface was 15.2 ± 3.21 mm.The coefficient variation assessing the anatomical reliability of different bony landmarks were as follows:ATFL fibular origination AA(17.54％) ＜AT(20.28％) ＜ AO(33.14％),CFL calcaneus insertion CC(17.70％)＜CP(18.57％)＜CS(21.1％).Conclusion Certain variations exist in the morphological parameters and the distances from the insertion of the lateral ankle ligaments to the adjacent bony landmarks.It provides anatomical evidence for lateral ankle ligament reconstruction in treating chronic ankle instability.