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AIM:Early calcification of atherosclerotic plaques are colocalized with macrophage and high mobility group box 1 (HMGB1), a cytokine associated with biomineralizing process under physiological and pathological conditions .Our study aims to evaluate whether HMGB1 induces ectopic mineralization via promoting the secretion of matrix vesicles ( MVs) from macrophages .METHODS:HMGB1 was added to the medium of macrophages , the secretion of MVs in the supernatant was tested by flow cytometry analysis .The mineral deposition in calcifying medium was detected by Alizarin Red staining and von Kossa staining .Transmission electron microscopy showed the formation of hydroxyapatite crystals in MVs .Then we subcutaneous injection into mice with MVs to induce regional minera-lization.RESULTS:HMGB1 significantly promoted secretion of MVs from macrophages as raveled by flow cytometry analysis .TNAP activity, considered as a marker of MVs maturation , was higher in HMGB1-induced MVs compared to the control-MVs.HMGB1-MVs also led to mineral deposition in an in vitro MVs-collagen mineralization model .Subcutaneous injection into mice with MVs derived from HMGB1-treated cells showed a greater potential to initiate regional mineralization .Mechanistic experiments revealed that HMGB 1 activated neutral sphingomyelinase 2 ( nSMase2 ) that involved the receptor for advanced glycation end products ( RAGE ) and p38 MAPK (upstream of nSMase2).Inhibition of nSMase2 with GW4869 or p38 MAPK with SB-239063 prevented MVs secretion and min-eral deposition .CONCLUSIONS: HMGB1 induces MVs secretion from macrophages at least in part , via the RAGE/p38 MAPK/nSMase2 signaling pathway .Our findings thus reveal a novel mechanism by which HMGB 1 may participated in the early calcification of atherosclerotic plaques .
ABSTRACT
[Objective]To research the efficient way of isolating and proliferating of induced endothelial cells(ECs) ,measure the number of the ECs and observe the main biological characteristic of ECs m vitro during the passage cultivation.[Method]The marrow mononuclear cells(MNCs) in rabbit marrow were isolated by the gradiem centrifugation and cultivated to isolate and induce ECs. The efficiency of isolating and harvesting ECs was measured by the number of the first passage ECs from every 10~6 of the MNCs. The changes of ECs morphology, growth and proliferation were observed during the passage cultivation. The immunohistochemical staining of CD31 and vWF and function of ECs were checked.[Result]After cultivated at 1?10~6/cm~2 MNCs for 7～8 d, the first passage ECs were harvested. The number of the first and the second passage ECs had a remarkable statistical positive correlation with the number of the MNCs from one rabbit. After cultivated for 7～8 d, the first passage ECs of 7.086?0.75?10~4 in number and the second passage ECs of 53.20?6.47?10~4 in number were harvested from every 10~6 MNCs on average;ECs, which were monolayer arrayed to form cobblelike shape,The cells of 2nd passage showed positive staining of CD31 and vWF, and positive labeling of DiI-LDL and UEA.[Conclusion]The period from cultivating to harvesting ECs is shortened at the density of 1?10~6/cm~2 MNCs. It is feasible that the efficiency of isolating and harvesting ECs could be measured by the number of the first or the second passage ECs from every 10~6 MNCs. The induced ECs can act as seeding cells in bone tissue engineering in future.
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Objective: To observe the clinical effect of integrated traditional and western medicine in treating chronic idiopathic thrombocytopenic purpura (CITP) elderly patients. Methods: Sixty patients were divided randomly into the treatment group (n=30) and the control group (n=30), the former was treated with danazol combining with prednisone and Chinese drugs, while the latter was treated with VCR. Results: The total effective rate of the treatment group was better than that of the control group, the difference between those two groups was significant (P
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Since the 1990s,models of genetically modified mice(gene knockout,mutation or transgene overexpression) have been increasingly used in heart research.The findings from this class of murine models have contributed significantly to a number of research breakthroughs achieved in the last 15 years.Currently,the number of genetically modified mouse strains has been increasing steadily.These models that targeted a family or a functionally relevant group of proteins have formed systemic tools for research use.The database of mouse cardiac phenotype has been established.Meanwhile,the research methodology and strategic approach exploring mouse cardiac phentypes at molecular to in vivo levels have been mature.It is expected that the use of this class of genetically targeted mouse models will become increasingly important in future research.
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Following a brief introduction of relaxin family members, receptors and related signalling pathways,we review effects of relaxin on cardiovascular system,particularly,its anti-fibrotic action.The heart and vessels are target organs of relaxin.Recent studies have documented that the relaxin genes are upregulated in the diseased heart of humans and experimental animals.In several animal disease models,relaxin can reverse cardiac fibrosis.Relaxin may also indirectly promote myocardial regeneration and repair by suppressing fibrotic healing.Further research needs to focus on elucidating the exact mechanisms utilized by relaxin to induce these cardiac actions and translation of experimental findings to novel therapy of cardiovascular disease.