1.Effects of different inner face of human venous blood container on platelet activation
Yingchun ZHOU ; Xifeng TANG ; Mingfei XU
Chinese Journal of Laboratory Medicine 2008;31(8):937-941
Objective To study the effects of different inner face status of human venous blood container on platelet activation. Methods The plastic( polyethene terephthalate,PET) and glass tubes were coated with polyalkyleneoxide modified polydimethylsiloxane(L722). The contact angles of L722-coated glass and L722- coated PET tubes, glass tubes, PET tubes, silane coupling agent-coated glass tubes and polypropylene (PP) tubes were analyzed respectively. The blood were drawn into above tubes, and then incubated in a roller bottle at room temperature for 10-60 mira The marker of activated platelete, CD62p, was detected by flow cytometry(FCM). Results The inner face contact angle of the blood collection tubes with different material and surface treatment reflected platelet activation to a certain extent, but was not linear. The percent age of CD62p positive platelets in L722-modified PET tubes reduced from (37.4 ±14. 8) % to(21.9 ± 12. 4) %. The platelet activation by glass tubes was (54.5 ± 18.6 ) %, markedly more than PET tubes. While membrane formed, the platelet activation by glass tubes decreased remarkably, and the percentage of CD62p positive platelet in silane coupling agent-coated glass tubes were ( 28. 3 ± 8.2 ) %,markedly less than that of the L722-coated glass tubes. The platelet activation by PET-based material ectally modified with L722 was obviously less than L722-coated glass tubes and the percentage of CD62p positive platelet in silane coupling agent-coated glass tubes were ( 41.5 ± 15.9 ) % and ( 22. 0 ± 12. 8 ) %,respectively. The time course of platelet activation by different tubes showed that the platelet activation by L722-coated PET tubes and polypropylene tubes in 60 rain was not significantly different from the results in30 mitt Conclusions The diverse surface energy status induced by different material and surface treatment of blood collection tubes have obvious effects on the activation of platelets. The silicic oil surface treatment can effectively improve the blood compatibility of blood collection tubes. CD62p detectod by FCM is a sensitive marker for the evaluation of the activation of platelets induced by the material of blood collection tubes. It is of importance in the establishment of surface treatment model of blood container and screening out the material of clinical application.
2."Basic ideas and structure of health services industry development in ""the 13th five-year plan"" period in Shanghai"
Yimin ZHANG ; Chen FU ; Yanting LI ; Hong LIANG ; Lingfang WU ; Mingfei XU ; Jiquan LOU ; Shanshan LIU
Chinese Journal of Hospital Administration 2015;(10):791-795
On the basis of analysis of the current status and future tendency of development of the health services industry in Shanghai,the authors identified key problems and bottlenecks.Thus they made clear the target positioning and principles of the industry,and proposed the basic ideas and pattern of the industry in the 13 th five-year plan period,focusing on such fields as private and high-end healthcare services,traditional Chinese medicine services,public health services,commercial health insurance,and other related industries.In the end,corresponding supporting polices were proposed.
3.Research progress of feature selection and machine learning methods for mass spectrometry-based protein biomarker discovery.
Kaikun XU ; Mingfei HAN ; Chuanxi HUANG ; Cheng CHANG ; Yunping ZHU
Chinese Journal of Biotechnology 2019;35(9):1619-1632
With the development of mass spectrometry technologies and bioinformatics analysis algorithms, disease research-driven human proteome project (HPP) is advancing rapidly. Protein biomarkers play critical roles in clinical applications and the biomarker discovery strategies and methods have become one of research hotspots. Feature selection and machine learning methods have good effects on solving the "dimensionality" and "sparsity" problems of proteomics data, which have been widely used in the discovery of protein biomarkers. Here, we systematically review the strategy of protein biomarker discovery and the frequently-used machine learning methods. Also, the review illustrates the prospects and limitations of deep learning in this field. It is aimed at providing a valuable reference for corresponding researchers.
Algorithms
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Biomarkers
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Humans
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Machine Learning
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Mass Spectrometry
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Proteomics
4.EFHD2 regulates the NOX4/ROS pathway to initiate glucose metabolism reprogramming and promote breast cancer progression
Mingfei XU ; Yi ZHANG ; Ruyi KANG ; Chaoyue LIU
Chinese Journal of Comparative Medicine 2024;34(9):66-75
Objective To investigate the mechanism by which EFHD2 affects the occurrence and progression of breast cancer based on the NOX4/ROS signaling pathway.Methods Cells were divided into an NC-shRNA group and EFHD2-shRNA group.A lentiviral vector for EFHD2 silencing and a control vector were constructed and used to transfect MDA-MB-23 and MCF-7 breast cancer cells.The transfection efficiency was verified by qRT-PCR.A CCK8 assay was used to detect cell proliferation activity.A plate cloning assay was employed to measure cell colony formation ability.A scratch test was used to detect cell migration,and a Transwell assay used to assess cell invasion.Flow cytometry was applied to detect apoptosis and ROS levels.qRT-PCR was used to analyze the mRNA expression of GLUT1,PDK1,PFK1,PKM2,PDH,and LDH,while Western blot was applied to detect the expression of Cleaved caspase-3,MMP-2,and NOX4 proteins.Results Compared with the NC-shRNA group,the EFHD2-shRNA group's EFHD2 expression was significantly decreased and its cell survival and colony formation ability were weakened.The apoptosis rate and the expression of the pro-apoptotic protein Cleaved caspase-3 increased.The cell migration distance was shortened,while the number of invading cells and the expression of MMP-2,which promotes migration and invasion,were decreased.The levels of lactic acid and GLUT1,PDK1,PFK1,PKM2,and LDH decreased,while the levels of ATP and PDH increased.Streaming result showed that ROS levels were reduced and NOX4 protein was down-regulated after silencing EFHD2.Conclusions EFHD2 inhibits ROS production by regulating the NOX4/ROS signaling pathway,causing lactic acid and glucose accumulation,promoting the apoptosis of breast cancer cells,and inhibiting cell proliferation,migration,and invasion.
5.Effects of pyrimidine bundle-binding protein-associated splicing factors on the function of hypoxia-induced human retinal microvascular endothelial cells
Manhong XU ; Linni WANG ; Tingting LIN ; Xinjun REN ; Yifeng KE ; Liying HU ; Mingfei JIAO ; Yong WANG ; Qiong WANG ; Yaru HONG ; Xiaorong LI ; Lijie DONG
Chinese Journal of Ocular Fundus Diseases 2020;36(2):135-142
Objective:To observe the effect of pyrimidine bundle-binding protein-associated splicing factors (PSF) on the function of hypoxia-induced human retinal microvascular endothelial cells (hRMECs).Methods:A three-plasmid system was used to construct lentivirus (LV)-PSF. After LV-PSF infected hRMECs in vitro, the infection efficiency was measured by flow cytometry. Real-time quantitative PCR (RT-PCR) was used to detect the expression of PSF mRNA in hRMECs infected with LV-PSF. The experiment was divided into two parts, in vivo and in vitro. In vivo experiments: 20 healthy C57B/L6 mice at the age of postnatal 7 were randomly divided into normal group, oxygen-induced retinopathy (OIR) group, OIR+LV-Vec group, and OIR+LV-PSF group, each group has five mice. Mice in 3 groups were constructed with OIR models except the normal group and the mice in OIR group were not treated. The mice in the OIR + LV-Vec group and the OIR+LV-PSF group were injected with an empty vector (LV-Vec) or LV-PSF in the vitreous cavity, respectively. The effect of LV-PSF on the formation of retinal neovascularization (RNV) was observed then. In vitro experiments: hRMECs were divided into normal group, hypoxia group, vector group, and PSF high expression group. HRMECs in the normal group were cultured in vitro; hRMECs in the hypoxic group were restored to normal culture conditions for 3 h after 3 h of hypoxia stimulation; hRMECs in the vector group and PSF high expression group were infected with LV-Vec and LV-PSF for 48 h, and hRMECs were returned to normal culture conditions for 24 h with hypoxia stimulation for 3 h. The effect of PSF on cell proliferation was observed by MTT colorimetry. Cell scratch test and Transwell migration experiment were used to observe the effect of PSF on cell migration ability under hypoxia stimulation. RT-PCR was used to observe the mRNA expression of HIF-1α, VEGF and PSF in each group of cells.Results:The LV-PSF of stably expressing PSF was successfully constructed. The infection efficiency was 97% determined by flow cytometry. The level of PSF mRNA in hRMECs infected with LV-PSF was significantly increased and detected by RT-PCR. In vivo experiments: The RNV area of the mice in the OIR group and the OIR + LV-Vec group was significantly increased compared to the normal group ( t=18.31, 43.71), and the RNV area of the mice in the OIR + LV-PSF group was smaller than that in the OIR group ( t=11.30) and OIR + The LV-Vec group ( t=15.47), and the differences were statistically significant ( P<0.05). In vitro experiments: MTT colorimetry results showed that the proliferative capacity of hRMECs in the hypoxic group was significantly enhanced compared with the normal group ( t=2.57), and the proliferative capacity of hRMECs in the PSF high expression group was significantly lower than that of the normal, hypoxic, and vector groups ( t=5.26, 5.46, 3.73), the differences were statistically significant ( P<0.05). The results of cell scratch test showed that the hRMECs could be stimulated by the hypoxia stimulation for 3 hours to restore the normal condition for 24 hours or 48 hours ( t=8.35, 13.84; P<0.05). Compared with the vector group, cell migration rate in the PSF-high expression group was not significant ( t=10.99, 18.27, 9.75, 8.93, 26.94, 7.01; P<0.05). Transwell experiments showed that the number of cells stained on the microporous membrane was higher in the normal group and the vector groups, while the number of cells stained in the PSF high expression group was significantly reduced ( t=9.33, 6.15; P<0.05). The results of RT-PCR showed that the mRNA expression of HIF-1α and VEGF in hRMECs in the hypoxic and vector groups increased significantly compared with the normal group ( t=15.23, 21.09; P<0.05), but no change in the mRNA expression of PSF ( t=0.12, 2.15; P>0.05); compared with the hypoxia group and the vector group, the HIF-1α and VEGF mRNA expression in hRMECs in the PSF high expression group were significantly decreased ( t=10.18, 13.10; P<0.05), but the PSF mRNA expression increased ( t=65.00, 85.79; P<0.05). Conclusion:PSF can reduce the RNV area in OIR model mice. PSF may inhibit hypoxia-induced proliferation and migration of hRMECs through the HIF-1α/VEGF signaling pathway.