ObjectiveTo investigate the role of sphingosine-1-phosphate （S1P） in abnormal ossification in ankylosing spondylitis （AS）， clarify the relationship between S1P and “angiogenesis-osteogenesis” coupling， and provide new strategies for AS treatment. MethodsFemoral heads from AS patients and patients undergoing routine hip replacement were collected for immunohistochemical （IHC） staining to evaluate osteogenesis and H-type vessel formation. In vitro， ELISA was used to quantify the synthesis of S1P and analyze the expression changes of S1P signaling pathway-related molecules during the osteogenic differentiation of mesenchymal stem cells derived from patients with ankylosing spondylitis （ASMSCs） and those from healthy donors （HDMSCs）， to evaluate the activation status of S1P pathway during osteogenesis. Sphingosine kinase 1 （SK1） expression was knocked down in MSCs， and the S1P receptor inhibitor FTY720 was applied to block S1P signaling. Alkaline phosphatase （ALP） activity and Alizarin Red S （ARS） quantification were used to assess the effect of S1P on ASMSCs osteogenesis. Conditioned medium from osteogenically induced MSCs was used to treat human umbilical vein endothelial cells （HUVECs） to evaluate the effect of S1P on angiogenesis. An AS mouse model （SKG mice） was treated with FTY720 or the SK1 inhibitor PF-543 citrate. IHC staining and micro-CT scanning were used to assess abnormal ossification and spinal fusion， and immunofluorescence was used to evaluate H-type vessel formation. ResultsCompared with Osteonecrosis of the Femoral Head（ONFH） patients， AS patients exhibited excessive osteogenesis and H-type vessel formation （OCN P＜0.001， CD31 P＜0.001， EMCN P＜0.001）. During osteogenic differentiation， S1P expression and secretion were significantly higher in ASMSCs than in HDMSCs （P=0.0179）. Inhibition of S1P signaling with FTY720 or SK1 knockdown significantly suppressed osteogenic differentiation （compared with ASMSC， ARS： HDMSC P=0.001 8， FTY720 P＜0.001， si-SK1 P＜0.001； ALP： HDMSC P=0.032 8， FTY720 P=0.001 6， si-SK1 P＜0.001） of ASMSCs and the angiogenesis of HUVEC（compared with ASMSC， cell-covered area， total loops， total tube length and total branch points P＜0.001）. Treatment with FTY720 or PF-543 markedly inhibited abnormal ossification and spinal fusion（compared with Curdlan， arthritis index score， P＜0.001； OCN：control P=0.002， PF-543 P=0.010 7， FTY720 P=0.015 9 ） in AS mice and reduced H-type vessel formation （CD31⁺EMCN⁺： compared with curdlan， control P＜0.001， PF-543 P=0.001 7， FTY720 P=0.002 1）. ConclusionIncreased S1P synthesis in ASMSCs promotes osteogenic differentiation via autocrine mechanisms and further enhances ossification by facilitating H-type angiogenesis. Inhibiting S1P secretion in ASMSCs significantly suppresses abnormal ossification in AS.

Intensive cancer treatment with drug combination is widely exploited in the clinic but suffers from inconsistent pharmacokinetics among different therapeutic agents.To overcome it,the emerging nanomedicine offers an unparalleled opportunity for encapsulating multiple drugs in a nano-carrier.Herein,a two-step super-assembled strategy was performed to unify the pharmacokinetics of a pep-tide and a small molecular compound.In this proof-of-concept study,the bioinformatics analysis firstly revealed the potential synergies towards hepatoma therapy for the associative inhibition of exportin 1(XPO1)and ataxia telangiectasia mutated-Rad3-related(ATR),and then a super-assembled nano-pill(gold nano drug carrier loaded AZD6738 and 97-110 amino acids of apoptin(AP)(AA@G))was con-structed through camouflaging AZD6738(ATR small-molecule inhibitor)-binding human serum albumin onto the AP-Au supramolecular nanoparticle.As expected,both in vitro and in vivo experiment results verified that the AA@G possessed extraordinary biocompatibility and enhanced therapeutic effect through inducing cell cycle arrest,promoting DNA damage and inhibiting DNA repair of hepatoma cell.This work not only provides a co-delivery strategy for intensive liver cancer treatment with the clinical translational potential,but develops a common approach to unify the pharmacokinetics of peptide and small-molecular compounds,thereby extending the scope of drugs for developing the advanced com-bination therapy.

Objective: To investigate the prevalence of adolescents dietary behavior in Shanghai, and to explore emotional influence on dietary behavior. Methods: A total of 7 456 students from 10 junior and 6 senior high schools in Shanghai were selected to participate in the questionnaire survey with the stratified random cluster sampling method. The survey included general information, eating behavior, PHQ-2 and GAD-7. Results: During the past week, the proportion of adolescents in Shanghai reported consumption of sugar sweetened beverages, sweet desserts, frequent fried food and fast food (≥4 times/week) were 13.26%, 16.90%, 6.99 % and 13.01%, respectively. The proportion of students reported consumption of fruits, vegetables, milk and breakfast every day were 56.96%, 73.00%, 65.03% and 76.11%, respectively. There were significant differences by sex and educational stages(both P <0.05). Adolescents with depression or anxiety have a higher incidence of unhealthy eating behaviors than those without depression or anxiety( P <0.01). After adjusting for gender, school, accommodation, grades, pocket money and social class, depression and anxiety increase the risk of various unhealthy eating behaviors in adolescents( P <0.05). Compared with those without anxiety, the risks of sugar sweetened beverages consumption (≥1 time/d) among adolescents with mild and moderate to severe anxiety were 1.42 times (95% CI =1.20-1.67) and 2.51 times (95% CI =2.09-3.01), the risks of insufficient fruits consumption (<1 time/d) were 1.30 times (95% CI =1.16-1.45) and 1.28 times (95% CI =1.11-1.47), the risks of insufficient vegetable consumption (<1 time/d) were 1.35 times (95% CI =1.20-1.52) and 1.41 times(95% CI =1.21-1.65), the risks of insufficient milk consumption (<1 time/d) were 1.29 times (95% CI =1.15-1.44) and 1.20 times(95% CI =1.04-1.39), and the risks of breakfast skipping were 1.75 times (95% CI =1.54-1.99) and 2.97 times (95% CI =2.55-3.46) among adolescents with mild and moderate to severe anxiety. Conclusion The proportion of unhealthy eating behaviors among adolescents in Shanghai is still high. Early education and intervention for students eating behaviors should be carried out, and attention should be paid to the occurrence of adolescents negative emotions, so as to reduce the risk of unhealthy eating behaviors among adolescents through the promotion of mental health.

Objective To observe the effect of hypoxia on the expression of epithelial growth factor receptor (EGFR) and cell apoptosis of breast and cervical cancer xenografts in nude mouse models.Methods The nude mouse models with MCF-7 and HeLa xenografts were established.The degree of hypoxia and EGFR expression were observed by confocal microscopy.The influence of EGFR expression on cell apoptosis under hypoxia was observed by TUNEL assay.Results EGFR expression was either up-regulated or down-regulated in the MCF-7 and HeLa cells with high degree of hypoxia.Furthermore,the degree of apoptosis was reduced in tumor tissues with high EGFR expression compared with that in those with low expression of EGFR.Conclusion The hypoxia in MCF-7 and HeLa cells exerts heterogeneous effect on EGFR expression.Under hypoxic conditions,EGFR exoression is negatively correlated with cell apoptosis.