1.Induced differentiation of human embryonic stem cells into endothelial cells
Yang SHI ; Junsheng SHEN ; Taotao WU ; Xiaofei LI
Chinese Journal of Tissue Engineering Research 2016;20(23):3394-3399
BACKGROUND:Human embryonic stem cels exhibit self-renewal and multi-differentiation potential, and can differentiate into endothelial cels under certaininduction conditions.
OBJECTIVE:To explore induced conditions of the human embryonic stem cels differentiating into endothelial cels and to investigate the effect of vascular endothelial growth factors on theendothelial differentiation of human embryonic stem cels.
METHODS:After resuscitation,passage40 human embryonic stem cel lines H9 weresubjected to suspension culture to prepare embryos, and after 5-day culture,these cels werecultured in attachment medium to differentiate into embryoid bodies,folowed by induction with50 μg/L vascular endothelial growth factors. Passage 2 and 15 embryonic stem cels after induced differentiation weretaken for Dil-Ac-LDL uptake test and immunohistochemical staining, respectively.
RESULTS AND CONCLUSION:After 1-day culture, cord-like or polygonal monolayer cels around embryoid bodies showed bud-like andradialgrowth witharelative rapid speed merging into surrounding colonies; at 2-3 days, the number of suspension cels increased further, but the smal-round cels in the center began to die; at 5 days, embryoid bodies started to passage, and aggregated cels exhibited typical paving stone-like appearance. Moreover, some human embryonic cels after induced differentiation could actively takeupfluorescent labeled LDL,andred fluorescent particlesappeared.Additionaly, passage 15 embryonic stem cels after induced differentiation could express CD31 and FLK-1.These findings suggest that human embryonic stem cels induced by vascular endothelial growth factors can differentiate into endothelial cels.
2.Effects of transfected gut-enriched Krüppel-like factor on growth of human gastric carcinoma cell line SGC-7901 and its xenograft in nude mice
Hui YAN ; Weichang CHEN ; Jiannong CEN ; Hongjie SHEN ; Xiaofei QI
Chinese Journal of Digestion 2011;31(1):30-35
Objective To investigate the antitumour effects of transfected gut-enriched Krüppellike factor(GKLF) on human gastric carcinoma cell line SGC-7901 in vitro and in vivo. Methods The expression of GKLF mRNA and protein in human gastric carcinoma cell line SGC-7901 were detected before and after transfection by real-time fluorescence quantitative PCR and Western blot,respectively. Proliferation and invasion in SGC-7901 were measured respectively by MTT assay, flow cytometry, colony formation assay and cell invasion assay after transfected with GKLF. The growth of xenograft was observed, the microvessel density(MVD) of xenograft tissue was determined by immunohistochemistry. Results The GKLF mRNA and protein in SGC-7901 were overexpressed after transfected with GKLF(P<0.05). The proliferative speed of SGC7901-pcDNA3.1-GKLF group was markedly lower than that of SGC-7901 and SGC7901-pcDNA3.1 groups (P<0.05). Transfected with GKLF caused part of the G0/G1 arrest, decreased clone formation rate and the invasion ability (P<0.05). The growth speed of xenograft in SGC7901-pcDNA3.1-GKLF group was lower, the weight and MVD of xenograft tissue in SGC7901-pcDNA3. 1-GKLF group were less (P< 0. 05).Conclusion Transfected with GKLF maysuppress proliferation and invasion in human gastric carcinoma cell line SGC-7901, inhibit the growth and the angiogenesis of xenograft in nude mice.
3.Relationship between BMD and Zn, Cu, Ca levels in the hair and meal in elderly people.
Wanli, LI ; Yuhui, TIAN ; Xiaofei, SONG ; Min, ZHANG ; Guanxin, SHEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2005;25(1):97-9
The relationship between bone mineral density (BMD) and Zn, Cu, Ca levels in the meal and hair of urban and rural elderly people were studied. 470 subjects above 60 years old (urban 205 and rural 265), 178 males with an average age of 65.70 +/- 3.48 and 292 females with an average age of 65.90 +/- 4.02, were inquired. The BMD and Zn, Cu, Ca levels in the meal and hair were measured. The detected BMD in urban and rural female old people was significantly lower than that of the males; The contents of Ca and Zn in the meal of the urban females were significantly lower than those of the urban males; The Ca, Zn in the meal and Zn in the hair of the rural females were significantly lower than those of rural males (P < 0.05 or 0.01). The BMD, Ca intakes, Ca and Zn in the hair of the rural old people were significantly lower than those of the urban old people (P < 0.05 or 0.01). There was a correlation between BMD with the Ca, Zn of the hair and dietary Ca, Zn, Cu or between dietary Zn with Ca, Zn in the hair and Ca, Cu intakes. The Zn, Cu and Ca levels in the meal nutrients were correlated with BMD to some degrees. Lack of Ca and Zn in the meal can cause the reduction of BMD.
Bone Density
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Cadmium/*analysis
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Copper/*analysis
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Diet Surveys
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Hair/*chemistry
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Nutritional Status
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Osteoporosis/prevention & control
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Rural Health
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Zinc/*analysis
4.Effects of testosterone on norepinephrine release in isolated rat heart.
Xiaofei, WANG ; Yanzhou, ZHANG ; Jun, BU ; Linghong, SHEN ; Ben, HE
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(1):42-6
The effects of testosterone on norepinephrine release were investigated in the isolated rat hearts. Sprague-Dawley male rats (n=120) were randomized to testosterone and control groups. The rats in testosterone group were perfused with modified Krebs-Henseleit buffer containing different concentrations of testosterone (0.1, 1.0, 10.0, and 100.0 nmol/L, respectively). Myocardial ischemia was induced by globally stopping the perfusion flow. Exocytotic norepinephrine release was induced by electrical field stimulation at 5 V (effective voltage) and 6 Hz (pulse width of 2 ms) for 1 min. The overflow of norepinephrine was determined by high pressure liquid chromatography and electrochemical detection (HPLC-EC). Following acute ischemia, testosterone (1.0, 10.0 and 100.0 nmol/L) significantly reduced norepinephrine release (P<0.01), and the norepinepherine overflow was similar between the control and 0.1 nmol/L testosterone group (P>0.05). Electrical stimulation of the ventricle evoked norepinepherine release, and this was diminished by the perfusion with testosterone at the concentrations of 1.0, 10.0 and 100.0 nmol/L (P<0.01). It is suggested that testosterone suppresses ischemia- and electrical stimulation-induced norepinepherine release in the isolated rat hearts.
5.Discussion on a new opened course combining complexity science and medicine in medical colleges
Li DING ; Jing LI ; Shen GU ; Xiaofei CHEN
Chinese Journal of Medical Education Research 2017;16(9):870-873
Complexity science is a new type of scientific research method, and the combination of complexity science, life science, and medicine is still in the initial stage of exploration, and domestic medi-cal colleges do not offer correlative course currently. In order to broaden students' vision, transform their traditional thinking model, and prepare the future of multidisciplinary cooperation, medical colleges should open complexity science and medicine correlative course for senior undergraduate and graduate students. Course content should start by the famous fablebutterfly infect, followed by the introduction of chaos and fractal, self-organization and complex system theory concept, theoretical framework, and their application in the medical field The teaching material should refer to some popular science books, such as Mystery of the butterfly effect. The teaching method should draw lessons from metaphor.
6.Decision-making under risk condition in benign epilepsy children with centrotemporal spikes
Xuehui HU ; Xiaocui WANG ; Liwei SHEN ; Xiaofei YE ; Bin YANG
Chinese Journal of Behavioral Medicine and Brain Science 2017;26(4):335-339
Objective To investigate the ability of decision-making under risk condition in benign epilepsy children with centrotemporal spikes (BECT).Method Seventy-eight BECT patients and seventyfive healthycontrols were studied by Iowa Gambling Task (IGT) with inexplicit probability and Game of Dice Test (GDT) with explicit probability.Results (1) There was a significant difference in IGT task scores between the two group (t=-2.56,P<0.05).The total number of advantageous choices in IGT were lower in BECT group (49.35±5.38) than that in healthy control group (52.24±6.23).BECT group (-1.41 ± 10.60) performed worse on task than healthy control group (4.43±11.88),and the difference was statistically significant (t=-3.21,P<0.05).The control group scored higher on Blockl,Block4 and Block5 than that of BECT group (P<0.05).(2) There was no significant difference between the BECT group (4.42±7.74) and the control group(5.49±6.81) in the GDT test scores (t=-0.91,P=0.37).(3) Pearson correlation analysis showed that there was no significant correlation between the age,age of onset,education,seizure frequency and onset duration and the score of IGT and GDT (r =-0.091-0.274,all P > 0.05).Conclusion There is no significant decision-making disability in children with BECT under the condition of certain risk probability,while there is significant impairment under the condition of uncertain risk probability.The decision-making disability may be associated with impaired frontal lobe function.
7.Changes of ACTH, Cs and PRL Levels in Peripheral Blood From Patients With Active Systemic Lupus Erythematosus
Xiaofei WANG ; Hui SHEN ; Xiaoli ZHANG ; Shuling BAI
Journal of China Medical University 2001;30(2):137-138,140
Objective: Our purpose was to investigate the changes of adrenocorticotropin (ACTH), Cortisol (Cs) and prolactin (PRL) levels in peripheral blood of patients with active systemie lupus erythematosus(SLE). Methods: Radioimmunoassy. Results: Active SLE patients had a low level of ACTH,Cs in peak phase and a high level of PRL. Consequentaly, the ratio of PRL/Cs increased, which was positively correlative to active index of SLE. Conclusion: Endogenetic secretion in sufficiency of glucocorticalsteroid and increase of PRL have important function in the initiation and development of SLE.
8.Glutamine regulates the proliferation and survival of small cell lung cancer H446 cells
Pengyu XU ; Jiayin LI ; Yajing MIAO ; Cuicui GAO ; Yao SHEN ; Fang JIN ; Xiaofei QIU
Tianjin Medical Journal 2016;44(12):1423-1427
Objective To investigate the effects of glutamine (Gln) on proliferation and survival of small cell lung cancer H446 cells, and further to explore the potential mechanism. Methods The proliferation of H446 cells was detected at different time points (0, 24, 48, 72 and 96 h) by CCK-8 assay in Gln (+) group and Gln (-) group, and an optimal time was selected. Under the optimal time, Annexin V-FITC/PI staining, CellTiter-Glo? assay kit and flow cytometer were used to detect cell survival, cellular adenosine triphosphate (ATP) and reactive oxygen species (ROS) levels. Gln (-) group was used as the control group, under the condition of Gln deficiency, cellular ATP, cell proliferation and survival were detected after adding oxaloacetic acid (OAA) or dimethyl-α-ketoglutarate (DM-αKG). Gln (-) group was used as the control group, cellular ROS, cell proliferation, colony and survival were detected after treated with ROS scavenger N- acetyl cysteine (NAC). With different concentrations (0, 2, 5, 10 μmol/L) of glutaminase inhibitor BPTES, the optimal concentration was selected through the colony assay. The cellular ATP and ROS levels and cell proliferation were detected under the optimal concentration. H446 cells were treated with bis-2-(5-phenylacetamido-1,2,4-thiadiazol-2-yl) ethyl sulfide (BPTES), ROS inducer hydrogen peroxide (H2O2) or the combination of them, and cell survival ratio was compared between two groups. Results The proliferation levels of H446 cells at 24, 48, which were decreased most significantly in 72 h in Gln (-) group. When 72 h was used as the optimal time, the cell survival ratio and ATP level were decreased, and the ROS level was increased, in Gln (-) group compared with those of Gln (+) group (P<0.05). There was a higher survival ratio in H446 cells in Gln (-)+OAA group and Gln (-)+DM-αKG group than that of Gln (-) group (P<0.05), but there were no significant differences in cell proliferation and ATP levels between Gln (-) group, Gln (-)+OAA group and Gln (-)+DM-αKG group. The ROS level was reduced, the cell proliferation, colony level and survival ratio were increased in Gln (-)+NAC group compared with those of Gln (-) group (P<0.05). Cloning assay showed that 10μmol/L was the optional concentration. Under this concentration, the proliferation and ATP level were decreased in Gln(+)+BPTES group (P<0.05), and cellular ROS level was up-regulated compared with Gln(+) group. The survival ratio was significantly lower in BPTES+H 2O2 group compared with BPTES (+) group or H2O2 (+) group. Conclusion Glutamine deficiency inhibits the proliferation and survival ratio of H446 cells through enhancing ROS level. BPTES and H2O2 show synergistically inhibitory effect on the survival of H446 cells.
9.Early efficacy study of matrix-induced autologous chondrocyte implantation repairing knee joint cartilage injury
Qing WANG ; Huayang HUANG ; Tao ZHANG ; Xiaofei ZHENG ; Pingyue LI ; Hongyuan SHEN ; Jiarong CHEN
Chinese Journal of Orthopaedics 2016;(1):28-34
Objective To study the feasibility and early efficacy of matrix?induced autologous chondrocyte implantation repairing knee joint cartilage injury. Methods The Matrix?induced autologous chondrocyte implantation was used to repair knee joint cartilage injury in 13 cases (11 males and 2 females) with knee joint cartilage injury from April 2012 to March 2013. The av?erage age was 27.5 years old. All cases were suffering from unilateral focal cartilage defect of knee joint with International Carti?lage Repair Society (ICRS) chondral defect classification system grade III or IV, visual analogue scale (VAS)>3, and all of which had corresponding pain symptoms. The average defect area was 4.2 cm2. Standardized rehabilitation exercise was carried out after matrix?induced autologous chondrocyte implantation. Patients were followed up for 1 years, and knee injury and use osteoarthritis outcome score(KOOS), International Knee Documentation Committee (IKDC), subjective knee form and Lysholm score were col?lected to assess the function. Meanwhile, magnetic resonance observation of cartilage repair tissue (MOCART) score was used to assess the magnetic resonance imaging. Results All patients had been followed?up for 1 year. One patient had meniscus repair under arthroscopy for the meniscus injury caused by downstairs sprain in 6.5 months postoperative, so the score of 12 months post?operative was excluded. The knee range of motion was decreased in 3 months postoperative (123.1°±8.0°) compared to preopera?tive one (135.4°±5.7°), and has no difference in 6 months (136.1°±6.1°) and 12 months postoperative (135.1°±3.6°) compared to preoperative one. The 5 subsets of KOOS score were decreased in 3 months compared to preoperative one, and were significantly increased in 6 months and 12 months. The IKDC has no difference in 3 months (26.1±3.9) compared to preoperative one (43.5± 6.5), and were significantly increased in 6 months (53.3±5.8) and 12 months (62.8±7.2) compared to preoperative one. The magnet?ic resonance observation of cartilage repair tissue (MOCART) score was increased in 12 month(73.3±17.9)compared to preopera?tive one(51.5 ± 12.6). Conclusion MACI is a good technology for knee joint cartilage injury. It has a good clinical effect on re? pairing cartilage injury effectively and restoring the function of knee joint.
10.Attention networks in benign epilepsy children with centrotemporal spikes
Xiaocui WANG ; Bin YANG ; Liwei SHEN ; Guange YANG ; Xiaofei YE ; Zhiming ZHANG
Chinese Journal of Neurology 2015;48(5):406-410
Objective To investigate the characteristics of attention networks impairment in benign epilepsy children with centrotemporal spikes.Methods The attention network test was performed in benign epilepsy children with centrotemporal spikes (n =75) and normal controls (n =75) on the efficiency of three anatomically defined attention networks:alerting,orienting,and execrtive control.Results The correct rate of attention network test was significantly lower in benign epilepsy children with centrotemporal spikes (95.77% ± 5.63%) than that in normal controls (98.55% ± 1.80% ; t =4.063,P < 0.05).The total average response time was significantly longer in benign epilepsy children with centrotemporal spikes ((946.87 ± 199.82) ms) than that in normal controls ((729.00 ± 146.75) ms ; t =-7.611,P < 0.05).The orienting network effect (29 (12,56)ms) was lower in benign epilepsy children with centrotemporal spikes than in normal controls (51 (21,78) ms; Z =-2.771,P =0.006).The executive control network effect was higher in benign epilepsy children with centrotemporal spikes (80 (50,105) ms) than in normal controls (57 (25,88) ms; Z =-2.146,P =0.032).However,the difference in alerting network effects between the two groups was not statistically significant (P > 0.05).Logistic regression analysis showed that the age on onset of the disease was an independent risk factor for the correct rate of attention network test and the total average response time (OR =1.830,95% CI 1.193-2.807,P =0.006 ; OR =1.635,95% CI 1.176-2.275,P =0.003).Conclusions The benign epilepsy children with centrotemporal spikes have impairment of attention networks.The age on onset of the disease has a significant impact on the attention of children with rolandic epilepsy.