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Objective:To investigate the expression of Galectin-1 in neuroblastoma(NB)tissues and the effects of down-regulating this gene on cell proliferation and invasion.Methods:A total of 62 cases of NB children who had complete data and were initially treated at the Department of Pediatrics, Yidu Central Hospital, Weifang Medical College from January 2010 to January 2018 were enrolled.The expression of Galectin-1 protein in NB tissues was detected by immunohistochemistry.NB cell line SH-SY5Y was cultured and divided into the siRNA-Gal1 group (with Galection-1 transfected interference sequence siRNA-Gal1), siRNA-NC group (negative control sequence siRNA-NC was transfected) and blank group(without any treatment). The expression of Galectin-1, cell proliferation activity, and cell migration and invasiveness in 3 groups were detected by real-time quantitative PCR, cell counting kit-8(CCK-8) assay and Transwell assay, respectively.Western blot were used to detect the expressions of Galectin-1, E-cadherin and Vimentin proteins in 3 groups.Results:Among 62 cases, the positive expression rate of Galectin-1 protein was 69.35%.The positive expression rates of Galectin-1 protein in cells with grade of clinicopathological indexes of International Neuroblastoma Staging System stage of Ⅲ-Ⅳ, cells at high risk and cells with bone metastasis (78.57%, 78.05% and 84.00%)were significantly different from stage of Ⅰ-Ⅱ and Ⅳs, cells at low risk and cells without bone metastasis (50.00%, 52.38%, 59.46%) (all P<0.05). Compared with the siRNA-NC group and the blank group, the expression of Galectin-1 mRNA(0.23±0.06 vs.1.04±0.05 and 1.00±0.08)and protein(0.23±0.05 vs.0.86±0.06 and 0.84±0.05)in the siRNA-Gal1 group was significantly decreased(all P<0.05). The cell optical density(OD)values at 24 h, 48 h, 72 h and 96 h in the siRNA-Gal1 group were significantly lower than those in the siRNA-NC group and the blank group(all P<0.05). Compared with the siRNA-NC group and the blank group, the siRNA-Gal1 group showed a significant decrease in cell migration(101.55±5.56 vs.137.24±5.14 and 132.76±7.46)and invasion(78.21±5.08 vs.114.46±7.31 and 120.06±6.47, all P<0.001). The expression level of Vimentin protein in the siRNA-Gal1 group was significantly lower than that in the siRNA-NC group and the blank group(0.24±0.03 vs.0.69±0.07 and 0.70±0.06)(all P<0.05), while the expression level of E-cadherin protein in the siRNA-Gal1 group was significantly higher than that in other 2 groups(0.77±0.09 vs.0.29±0.05 and 0.33±0.04)(all P<0.05). Conclusions:The positive expression rate of Galectin-1 protein in NB tissues is 69.35%, which indicates that Galectin-1 might be involved in the malignant process of NB.Silencing the expression of Galectin-1 gene can inhibit the proliferation, migration and invasion of NB cells, and the mechanism may be related the inhibition of epithelial-mesenchymal transition.
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Objective To investigate the expression changes of soluble urokinase-type plasminogen activator receptor(suPAR)and soluble triggering receptors expressed by myeloid cell-1(sTREM-1)in serum of children with primary nephrotic syndrome(PNS)and their clinical significance.Methods A total of 92 cases of newly diag-nosed PNS children were selected in Central Hospital of Yidu Affiliated to Weifang Medical College from June 2014 to September 2016.According to presence or absence of acute tubular necrosis,they were divided into acute renal injury group(27 cases)and non-acute renal injury group(65 cases).According to pathology type,they were divided into mesangial proliferative glomerulonephritis(30 cases),focal segmental glomerulosclerosis(23 cases),membranous ne-phropathy(18 cases),minimal change disease(14 cases)and membrane proliferative glomerulonephritis(7 cases).In the same period,45 healthy children were selected as the healthy control group.The clinical data were collected.The serum levels of suPAR and sTREM-1 were measured by adopting enzyme-linked immunosorbent assay(ELISA). Results The levels of total cholesterol(TC),triglycerides(TG),uric acid(UA),urinary protein/creatinine,24 h urinary protein,urinary N-acetyl-β-glucosaminidase(NAG)and β2-microglobulin(MG)in children with PNS were higher than those in the healthy control group,while serum albumin(ALB)was lower than that in the healthy con-trol group,and the differences were statistically significant(all P<0.05).The serum levels of suPAR and sTREM-1 in PNS patients were(133.09 ± 62.48)ng/L and(79.29 ± 34.68),respectively,which were significantly higher than those in the healthy control group[(31.11 ± 11.61)ng/L and(25.08 ± 8.10)ng/L](t=51.714,49.435;all P=0.000).The serum levels of suPAR and sTREM-1 in acute renal injury group were(188.82 ± 32.21)ng/L and (109.11 ± 24.78)ng/L,respectively,which were significantly higher than those in non -acute renal injury group [(75.96 ± 28.69)ng/L and(52.23 ± 14.07)ng/L]and healthy control group[(31.11 ± 11.61)ng/L and (25.08 ± 8.10)ng/L](F=16 739.607,10 487.256,all P=0.000).The serum levels of suPAR and sTREM-1 in children with focal segmental glomerulosclerosis and membrane proliferative glomerulonephritis were higher than those with minimal change disease,membranous nephropathy and mesangial proliferative glomerulonephritis,and the differences were statistically significant(all P<0.05).Pearson correlation analysis results showed that the serum levels of suPAR and sTREM -1 were positively correlated with TC,TG,urinary protein/creatinine,24 h urinary protein, urinary NAG and β2-MG(all P <0.05),while negatively correlated with ALB(P <0.05). Conclusions The serum levels of suPAR and sTREM-1 are elevated in children with PNS,and which are related with acute renal injury and pathological type,which can reflect the degree of renal tubular disease and kidney function to a certain extent.
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<p><b>OBJECTIVE</b>This study aims to determine the gene expression changes of iron transporters-divalent metal transporter 1 (DMT1) and ferroportin 1 (Fpn1) in the duodenal tissue of diet-induced obese mice.</p><p><b>METHODS</b>C57BL/6J mice were randomly divided into normal control (NC) and obesity model (OM) group, 6 in each, and fed on conventional and high-fat diet respectively for 14 weeks by table of random number. Then the DMT1 and Fpn1 mRNA contents in duodenal tissues of the animals were measured by Real-time PCR method, and the protein expression levels were analyzed by Western blot test.</p><p><b>RESULTS</b>The Real-time PCR detection results showed that, compared with the NC group for which the mRNA expression level was defined as 1.0, the Fpn1 mRNA expression in OM group (0.58±0.11) was reduced significantly (t = 6.71, P = 0.014), whereas the relative expression level of DMT1 mRNA in OM group (0.89±0.26) showed no obvious alteration (t = 2.01, P = 0.122). Western blot results showed that the relative protein expression levels of Fpn1 in OM and NC group were 0.32±0.06 and 0.65±0.19, respectively, and the difference was statistically significant (t = 5.37, P = 0.026). The DMT1 protein relative abundance was 0.88±0.21 in OM group and 0.92±0.17 in NC group, and the difference has no statistical significance (t = 1.84, P = 0.185).</p><p><b>CONCLUSION</b>Fpn1 gene expression is inhibited in the duodenum of diet-induced obesity mouse while DMT1 expression keeps unchanged, and this implies that decreased iron export from enterocytes into circulation might be responsible for the impaired iron absorption in obesity.</p>
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Animals , Mice , Cation Transport Proteins , Diet , Diet, High-Fat , Duodenum , Gene Expression , Iron , Mice, Inbred C57BL , Mice, Obese , RNA, MessengerABSTRACT
Objective To evaluate the implementation feasibility of the nurse performance appraisal and allocation plan based on the holistic nursing mode, and the clinical effect in mobilizing nursing staff's work enthusiasm and ensuring the quality of nursing service. Methods The nurse performance appraisal and allocation plan were further improved based on the previous research of this index system construction. The model were implemented in 32 clinical departments of Yidu Center Hospital of Weifang City. Four aspects data were collected for analysis six months later to evaluate the effect. They were nurses on merit pay distribution satisfaction, patients′satisfaction, doctors′satisfaction with nursing job, different clinical departments′nursing quality scores. Results Six months later, four factor scores of nurses to the satisfaction of merit pay distribution were significantly improved. The scores of fairness and impartiality evaluation of the merit pay distribution, incentive effect evaluation, pay and return on equity evaluation and performance pay gap rationality evaluation were higher than before [(3.39 ±0.64) points vs. (1.88 ±0.33) points, (3.28 ±0.74) points vs. (1.84 ±0.49) points , (3.28 ±0.71) points vs. (1.88 ±0.42) points and (3.38 ±0.67) points vs. (2.01±0.53) points, t=19.28, 16.22, 18.08, 16.79, all P<0.05]. Patients′satisfaction, doctors′satisfaction with nursing job and different clinical department' nursing quality scores were significantly increased as well [(99.14±0.82) points vs. (96.78±0.84) points, (96.59±0.91) points vs. (93.59±1.27) points and(97.67±0.41) points vs. (95.70±1.13) points]. Difference had statistical significance (t=11.79, 11.63, 9.60, P<0.05). Conclusions The performance salary allocation plan can effectively improve the hospital nurses on performance salary allocation satisfaction, patients′satisfaction, doctor′s satisfaction with nursing work and the quality of nursing department, and has good incentive in arousing the work enthusiasm of nursing staff and in ensuring the quality of nursing service.
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Objective We established the animal models of obesity induced by high-fat diet, in order to study the mRNA and protein expression of regulation molecules related with iron metabolism about hepcidin, lipocalin-2 ( LCN2 ) , ferroportin-1 (FPN1) in obese mice’ s liver and the molecular regulation mechanism.Methods C57BL/6J (4 ~6 weeks) mice were randomly divided into control group and obesity model group, each group of ten.The obesity group were fed with a high-fat diet and the control group were given the normal diet for lasting 15 weeks.After we successfully established the obesity animal model, the expression level of hepcidin, LCN2 and FPN1 mRNA in the liver were measured by Real-time fluorescent quantitative PCR method and the protein expression level of LCN2 and FPN1 were measured by Western-Blot.Results Compared with the control group, the expression level of hepcidin mRNA in the liver was increased in obesity group (P 0.05).Conclusion Obesity can increase the expression of hepcidin mRNA, however, there was no significantly effect on the expression of LCN2, FPN1.So, we can’t think that obesity can affect the expression of LCN2 and FPN1, lead to the ability of cells uptake and release iron abnormal, then appear iron metabolism disorders.As a result, leading to iron deficiency.Maybe obesity can affect other regulatory molecules related with iron metabolism through up-regulation the expression of Hepcidin or the more complex regulatory mechanisms.We still need further experimental research and exploration.This research also provides the basis of theoretical and experimental for the further study the effects of obesity on the expression of regulation molecules related with iron metabolism in obesity mice’ s liver and the mechanism of iron deficiency.
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Objective To study the expression of divalent metal transporter 1(DMT1)and ferroportin 1(FPN1)in obese mice’ s duodenal epithelium and investigate the mechanism of the effect of obesity on iron absorption in mice. Methods C57BL/6J mice were randomly divided into control group and obesity model group, each group of 6, To establish obese mice model by having a high-fat diet and the control group were fed with a normal diet for 12 weeks.After completion of modeling, The level of DMT1 and FPN1 mRNA expression in the duodenum were measured by real-time fluorescent quantitative PCR( Real-time PCR) method, the protein expression of FPN1 was measured by Western-Blot. Results Compared with the control group, the level of DMT1、FPN1 mRNA and FPN1 protein expression in the duodenum were decreased significantly in obese mice ( P <0.05 ) .Conclusion Obesity can decrease the expression levels of DMT1、FPN1 mRNA and FPN1 protein and induce iron deficiency,in order to provide experimental and theoretical basis for studying the mechanism of iron deficiency caused by obesity further.
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@#ObjectiveTo establish a new method for preparing synaptosomes.MethodsDensity gradient centrifugation method was used to isolate synaptosomes of mouse, checking by transmission electron microscopy.ResultsSynaptosomes prepared by this method had intact morphological characteristics, surrounding with a continuous oval-shaped membrane structure, moreover, mitochondrion and lots of synaptic vesicle in them.ConclusionThis method is applicable to establish a rapid, convenient and useful method for preparing synaptosomes.
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@#ObjectiveTo explore the effects of high fat diet and caloric restriction on brain aging as well as the activity of Acetylcholinesterase(AChE) and afford scientific evidence to rational diet and prevent brain aging.MethodsSixty male ICR mice were randomly divided into 6 groups: the D-galactose-induced brain aging, brain aging plus high fat diet, brain aging plus caloric restriction, high fat diet only, caloric restriction only and normal control groups. Mice were given 100 mg/kg·d subcutaneous injection of D-galactose to prepare brain aging model for 9 weeks. Morris water maze (MWM) test was employed to determine their spatial learning and memory ability. Acetylcholinesterase (AChE) activity in brain was determined by hydroxylaminecolorimetric assay.ResultsIn Morris water maze test, brain aging mice showed a significant longer escape latency than the normal control mice (P<0.05). There was no statistical difference in escape latency between brain aging mice plus high fat diet and brain aging mice groups (P>0.05), and between the control and high fat diet groups (P>0.05). Brain aging mice plus caloric restriction exhibited a significant shorter escape latency than brain aging mice (P<0.05), but no difference was found when compared with normal control mice (P>0.05). There were no statistical difference in escape latency between the controls and caloric restriction group (P>0.05). The AChE activity in brain aging, brain aging plus high fat diet and brain aging plus caloric restriction group were higher than those in control and caloric restriction group (P<0.05). There were no statistical difference in AChE activity between the controls and caloric restriction group (P>0.05). Brain aging plus high fat diet were higher than brain aging and other non model control groups.ConclusionHigh fat diet can raise the activity of AChE effectively, but can not influence the capacity of learning and memory in mice. Caloric restriction can improve the capacity of learning and memory in mice, but has no significant influence on the activity of AChE in brain.
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Objective To explore the activity of Elemene for glioma cell from the cellular and molecular level. Methods The human glioma cell U251 was cultured. The effect of Elemene for human glioma cell proliferation was studied by MTT assay. Cell cycle, Fas, PCNA, bcl-2, intracellular Ca~(2+) and apoptosis were evaluated by flow cytometry analysis. Results Elemene exhibited antiproliferative effect on human glioma cell U251 markedly. The fifty percent inhibition on concentration (IC_(50)) of Elemene against glioma cells at different time points. 24 h was 40.60 μg/ml, the 48 h 38.14 μg/ml and the 72 h 34.35 μg/ml.Cell cycle was blocked in the S and G_2/M phases. The apoptosis ratio was increased by Annexin V staining markedly. Elemene decreased the gene expressions of PCNA and Fas, increased the intracellular Ca~(2+). There was no significant effect on the bcl -2 gene expression. Conclusion Elemene exhibits a marked antiproliferative effect on glioma cells and induces apoptosis by decreasing the expression of PCNA and increasing intracellular Ca~(2+). It also influences the expression of Fas. It might have no relationship with bcl-2 gene expression.
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@#Objective To observe effects for exercise and diet on learning and memory ability in mice with encephalon aging induced by D-galactose(D-gal).Methods The model of mice with encephalon aging was made by D-gal.The learning and memory ability of mice was determined by Morris water maze.Results There was significant difference between high fat feed encephalon aging group and restrict food on normal feed encephalon aging group,normal feed and exercise encephalon aging group,high fat feed and exercise encephalon aging group,high fat feed normal group,normal feed and exercise group,normal feed group(all P<0.05).In spatial probe test,there was significant difference between restrict food on normal feed encephalon aging group and normal feed encephalon aging group(all P<0.05).Conclusion Exercise and restrict food can improve the learning and memory function in the mice;feed with high fat can promote encephalon aging.