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Objective:To explore the effect and mechanism of small GTP-binding protein GDP dissociation stimulator (SmgGDS) on the development of obesity.Methods:(1) 8-week-old C57BL/6J mice were randomly assigned to normal diet and high fat diet group, with 6 mice in each group. They were fed regular feed and a high fat diet containing 60% fat for 4 months, respectively. The expression of SmgGDS in epididymal adipose tissue (eWAT), liver, and skeletal muscle were measured using Western-blot. (2) 6-week-old wild-type (WT) and SmgGDS knockdown (KD) mice were divided into four groups, each receiving high fat diet for 4 months (7 in each group) and 7 months (9 in each group). Glucose tolerance test (GTT) and insulin tolerance test (ITT) were conducted; the weight, adipose tissue, and liver weight of mice were recorded; HE staining examined adipose tissue structural changes; Western-blot determined extracellular signal-regulated kinase (ERK) 1/2 phosphorylation levels in eWAT; real time fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect mRNA levels of CCAAT/enhancer binding protein α (C/EBPα), C/EBPβ and peroxisome proliferator activated receptor γ (PPARγ) in eWAT. (3) Mouse embryonic fibroblasts (MEFs) extracted from WT and KD mice were induced for differentiation. Oil red O staining and Western-blot were used to detect lipid droplet and expression of SmgGDS and phospho-ERK; C/EBPα, C/EBPβ and PPARγ mRNA levels were measured using RT-qPCR. (4) 10-week-old C57BL/6J mice were randomly assigned into two groups, with 7 mice in each group. Mice were infected with SmgGDS overexpressing adeno-associated virus (AAV-SmgGDS) or empty vector intraperitoneally, then fed with high fat diet. After 4 weeks, performed GTT and ITT; recorded the weight and adipose tissue weight of mice; HE staining was used to analyze structural changes of eWAT; Western-blot was used to detect the phosphorylation level of ERK in eWAT.Results:(1) The expression of SmgGDS was significantly upregulated in eWAT of high fat diet fed mice (normal diet group: 0.218±0.037, high fat diet group:0.439±0.072, t=2.74, P=0.034). (2) At 4 months of high fat diet intervention, the glucose tolerance (60 minutes after glucose injection, WT group: 528 mg/dl±21 mg/dl, KD group: 435 mg/dl±17 mg/dl, t=3.47, P=0.030; 90 minutes, WT group: 463 mg/dl±24 mg/dl, KD group: 366 mg/dl±18 mg/dl, t=3.23, P=0.047;120 minutes, WT group: 416 mg/dl±21 mg/dl, KD group: 297 mg/dl±16 mg/dl, t=4.49, P=0.005) and insulin sensitivity (15 minutes after insulin injection, WT group: 77.79%±3.45%, KD group: 54.30%±2.92%, t=3.49, P=0.005; 30 minutes, WT group: 62.27%±5.31%, KD group: 42.25%±1.85%, t=2.98, P=0.024; 90 minutes, WT group: 85.69%±6.63%, KD group: 64.71%±5.41%, t=3.12, P=0.016) of KD mice were significantly improved compared to the WT group, with an increase in eWAT weight ratio (WT: 4.19%±0.18%, KD: 5.12%±0.37%, t=2.28, P=0.042), but a decrease in average adipocyte area (WT group: 5 221 μm2±241 μm2, KD group: 4 410 μm2±196 μm2, t=2.61, P=0.026). After 7 months of high fat diet, the eWAT weight ratio of KD mice decreased (WT: 5.02%±0.20%, KD: 3.88%±0.21%, t=3.92, P=0.001) and adipocyte size decreased (WT group: 6 783 μm2±390 μm2, KD group: 4 785 μm2±303 μm2, t=4.05, P=0.002). The phospho-ERK1 in eWAT increased (WT group: 0.174±0.056, KD group: 0.588±0.147, t=2.64, P=0.025), and mRNA level of PPARγ significantly decreased (WT group: 1.018±0.128, KD group: 0.029±0.015, t=7.70, P=0.015). (3) The expression of SmgGDS was significantly increased in differentiated MEF (undifferentiated: 6.789±0.511, differentiated: 10.170±0.523, t=4.63, P=0.010); SmgGDS knock-down inhibited lipid droplet formation in MEF (WT group: 1.00±0.02, KD group: 0.88±0.02, t=5.05, P=0.007) and increased ERK1 (WT group: 0.600±0.179, KD group: 1.325±0.102, t=3.52, P=0.025) and ERK2 (WT group: 2.179±0.687, KD group: 5.200±0.814, t=2.84, P=0.047) activity, which can be reversed by ERK1/2 inhibitor. (4) SmgGDS over expression resulted in weight gain, increased eWAT weight (control group: 3.29%±0.36%, AAV-SmgGDS group: 4.27%±0.26%, t=2.20, P=0.048) and adipocyte size (control group: 3 525 μm2±454 μm2, AAV-SmgGDS group: 5 326 μm2±655 μm2, t=2.26, P=0.047), impaired insulin sensitivity(30 minutes after insulin injection, control group: 44.03%±4.29%, AAV-SmgGDS group: 62.70%±2.81%, t=3.06, P=0.019), and decreased ERK1 (control group: 0.829±0.077, AAV-SmgGDS group: 0.326±0.036, t=5.96, P=0.001) and ERK2 (control group: 5.748±0.287, AAV-SmgGDS group: 2.999±0.845, t=3.08, P=0.022) activity in eWAT. Conclusion:SmgGDS knockdown improves obesity related glucose metabolism disorder by inhibiting adipogenesis and adipose tissue hypertrophy, which is associated with ERK activation.
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Objective Recent evidence has implicated Larp1,an RNA-binding protein,in cell motility and EMT therefore prompting the study of Larp1 in EOC.This project aims to examine the potential role of Larp1 in cell invasion.Methods Ovarian cancer cells SKOV-3 and OVCAR-3 were transfected with DNA constructs to overexpress Larp1 and transfect cells were used to assess the overexpression of Larp1 in cell invasion and metastasis using several functional assay.Results Larp1 overexpression in both OVCAR-3 and SKOV-3 cells appear to enhance the invasive ability of cells and the accumulation of Larp 1 in these chemoinvasive protrusions strongly suggests the potential role of Larp1 in invadopodia formation.Cell spot chemoinvasion assay demonstrated increased chemoinvasion of Larp1 overexpressing SKOV-3 and OVCAR-3 cells towards a gradient of TGF-β,EGF and bFGF.Conclusions With its potential role in EMT and cell invasion,a deeper understanding of the physiological role of Larp1 in cancer cell motility will be potentially beneficial in the diagnosis and treatments of ovarian cancer.
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Objective@#To investigate the effects of apolipoprotein E deficiency (Apo E-/-) on plasma and lipoprotein distribution of sphingosine-1-phosphate (S1P) in mice.@*Methods@#Five male or female Apo E-/- or wild type (WT) mice were fed with chow diet and sacrificed at 32-week-age and plasma was collected. The constituents of lipoprotein(very low density lipoprotein (VLDL), low density lipoprotein (LDL), high density lipoprotein (HDL)) were separated by ultracentrifuge. The protein concentration of constituents was detected by BCA protein quantitative kit, and the S1P concentration in plasma and various lipoprotein constituents was detected by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Western blot was used to determine the plasma, liver, and kidney protein expression of apolipoprotein M(Apo M), which is considered as specific ligand of S1P.The S1P concentration in plasma and various constituents of lipoprotein in the Apo E-/- mice was compared to respective WT mice.@*Results@#(1)Plasma S1P content was significantly higher in the Apo E-/- groups than that of WT groups (male: (535.7±78.5)nmol/L vs. (263.3±22.0)nmol/L; female: (601.1±64.0)nmol/L vs. (279.0±33.9)nmol/L; all P<0.01). (2) Compared with WT mice, S1P content in non-HDL(LDL+ VLDL) was significantly higher in Apo E-/- mice (male: (504.9±52.8)nmol/L vs. (28.7±9.0)nmol/L; female: (427.7±27.4) vs. (27.8±4.7)nmol/L; after standardization of protein concentration, male: (385.0±41.2)pmol/mg protein vs. (71.4±6.6)pmol/mg protein; female: (330.2±22.0)pmol/mg protein vs. (67.2±12.1)pmol/mg protein; all P<0.01). (3) The expression of Apo M in plasma, liver and kidney was significantly higher in Apo E-/- groups than that of WT groups(all P<0.05).@*Conclusion@#The deficiency of Apo E could lead to upregulated S1P expression in the non-HDL, the underlying mechanism might be the increased transfer of HDL into the non-HDL by Apo M-S1P.
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AIM:To investigate the effect of quercetin on the biological functions of rat bone marrow-derived endothelial progenitor cells (EPCs) and its potential mechanisms.METHODS:The bone marrow-derived mononuclear cells of Sprague-Dawley rats were isolated by density gradient centrifugation.The differentiated EPCs were cultured specially and stained with DiI-Ac-LDL and FITC-UEA-1.CD133+ and FLK-1+ were detected on the cell surfaces.After 14 d, the EPCs were incubated with a PI3K inhibitor BYL719 (3 μmol/L) and an ERK inhibitor FR180204 (15 μmol/L).After incubation of the inhibitors for 2 h, the cells were treated with quercetin at different concentrations (0, 10, 20, 40, 80 and 100 μmol/L).MTT assay and Transwell assay were used to detect cell viability and the number of migratory cells.The protein levels of AKT, eNOS, ERK and their phosphorylated status were determined by Western blot.RESULTS:Quercetin enhanced the viability and migration of the EPCs at a dose-dependent manner.However, the PI3K inhibitor BYL719 suppressed the QUE-induced cell viability and migration.Moreover, ERK inhibitor FR180204 exerted the similar inhibitory effect on the cell viability but had no effect on cell migration.Quercetin activated the phosphorylation of AKT, eNOS and ERK.On the other hand, BYL719 was observed to inhibit the phosphorylation of AKT and ERK.FR180204, however, was showed to inhibit the phosphorylation of ERK only.On the contrast, the stimulatory effects that quercetin exerted on the expression of eNOS and its phosphorylation were suppressed by BYL719 and FR180204.CONCLUSION:Quercetin stimulates the viability and migration of EPCs via PI3K/AKT/eNOS and ERK/eNOS signaling pathway, which would be beneficial for cardiovascular health.
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Objective To investigate the role of the activated rhuPAa-melittin in ovarian cancer cells and to study the inhibitory effect of rhuPAa-melittinon on ovarian cancer cells. Methods rhuPAa-melittin was used to treat the ovarian cancer cells at different concentrations for 48 hrs. Then flow cytometery was applied to detect the cell cycle and cell apoptosis. rhuPAa-melittin protein was delivered to the mouse mode to investigate the effect of rhuPAa-melittin on the growth of the xenotransplanted tumor. Results rhuPAa-melittin was used to treat ovarian cancer cells at the concentration of 0 ,4 ,8 and 16 μg/mL for 48 hrs ,respectively. The results of cell apoptosis assay was 1.16%,3.83%,6.51% and 10.2%,respectively. Moreover,different concentrations of rhuPAa-melittin had no effects on the cells at G0/G1 phase,rhuPAa-melittin inhibited S phase cells to process into G2/M phase, contributing to suppressing the growth of ovarian cancer cells. Conclusion The in vivo and in vitro studies revealed that rhuPAa-melittin inhibits the growth of ovarian cancer.
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Objective To investigate the role of the activated rhuPAa-melittin in ovarian cancer cells and to study the inhibitory effect of rhuPAa-melittinon on ovarian cancer cells. Methods rhuPAa-melittin was used to treat the ovarian cancer cells at different concentrations for 48 hrs. Then flow cytometery was applied to detect the cell cycle and cell apoptosis. rhuPAa-melittin protein was delivered to the mouse mode to investigate the effect of rhuPAa-melittin on the growth of the xenotransplanted tumor. Results rhuPAa-melittin was used to treat ovarian cancer cells at the concentration of 0 ,4 ,8 and 16 μg/mL for 48 hrs ,respectively. The results of cell apoptosis assay was 1.16%,3.83%,6.51% and 10.2%,respectively. Moreover,different concentrations of rhuPAa-melittin had no effects on the cells at G0/G1 phase,rhuPAa-melittin inhibited S phase cells to process into G2/M phase, contributing to suppressing the growth of ovarian cancer cells. Conclusion The in vivo and in vitro studies revealed that rhuPAa-melittin inhibits the growth of ovarian cancer.
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Objective To construct a highly efficient expression plasmid of eukaryotic nuclear membrane protein Omega 3 fatty acid desaturase gene Fat‐1 in E .coli .Methods Using molecular cloning technology to construct the recombinant prokaryotic expression plasmid pET32a Fat‐1 and pET32a‐Mistic‐Fat‐1 fused with Membrane proteins expression chaperon mistic ;the two re‐combinant plasmids were transformed into E .coli strain BL21 (DE3) ,the expression of Fat‐1 protein and M110 Fat‐1 protein in‐duced by IPTG were identified by SDS‐PAGE and gray degree analysed the amount of expression ,further identified by Western blot .Results The results of enzyme digestion and sequencing demonstrated that we successfully constructed the prokaryotic ex‐pression vectors pET32a Fat‐1 and pET32a‐Mistic‐Fat‐1;SDS‐PAGE and Western blot showed that Fat‐1 fatty acid desaturase wasn′t significantly induced ,but the overexpression of M110 Fat‐1 fusion protein was obtained in E .coli ,accounting for 15% of the total amount of whole cell proteins .Conclusion The fusion with Mistic proteins to express the Fat‐1 gene has realized the overex‐pression of eukaryotic nuclear membrane integrated protein Omega 3 fatty acid desaturase in prokaryotic host .
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Background and Objective: Trimetazidine has mainly been used in coronary insufficiency, angina and elderly myocardial infarction. However, the effect of trimetazidine on the efficacy, heart rate variability [HRV] and protection of myocardial ischemia in elderly patients with acute coronary syndrome [ACS] remains unclear. This study aimed to investigate the effect of trimetazidine on the efficacy HRV and protection of myocardial ischemia in patients with ACS
Methods: One hundred twenty two elderly ACS patients who were above 70 years were chosen and randomly divided into two groups. One group was given conventional therapy, such as aspirin, isosorbide mononitrate and fluvastatin, and the other group was administered trimetazidine in addition to conventional therapy. The treatment period was eight weeks. A PI-2.22B three-channel AECG system was used on every patient for 24 hour dynamic electrocardiogram monitoring and HRV analyses on the first day after admission and eight weeks after treatment. HRV, 24 hour RR intermediate stage standard deviation [SDNN], five minutes average normal cardiac cycle standard deviation in 24 hour [SDANN], 24 hour close together normal cardiac cycle difference value mean square root [rMSSD], the percentage of difference of close together RR intermediate > 50 ms account total RR intermediate [PNN50], high frequency [HF] and low frequency [LF] parameters of patients were observed before and after treatment
Results: The SDNN, SDANN, rMSSD, PNN50 and HF parameters significantly increased compared with the conventional treatment group [all P < 0.05]. LF and LF/HF were significantly decreased in the trimetazidine treatment group compared with those in the conventional treatment group [all P < 0.05]
Conclusion: Trimetazidine improves HRV of elderly ACS patients and reduces cardiovascular events
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Science and technology innovation is an important quality of young talents in the new period requirements and training objectives,as well as hospital's driving force and source of the future sustainable development.Under the innovation-driven strategy,characterized by cultivating young talents of science and technology innovation,with the efforts to develop and improve young talents cultivation way,our hospital aim to accelerate the development of young talent as soon as possible.By setting up a hospital's fund,we gradually formed a stable funding system including the four directions of clinical,nursing,management,education.With the development purposes of young researchers as the main body,adhere to improve the quality of hospital funds is the key,we aim at forming a reasonable fund scale.Through the top level design,strict process management,implementation of opening argument and a series of strong measures to ensure project quality,we promote youth science and technology innovation talents effectively.
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Application of the theory of PDCA cycle to guide to build the hospital's project continuous quality improvement management mode,By Analyzing reasons,optimizing processes,improving the bidding system,regulating the declaration format,increasing of directors' participation,playing the role of expert,and evaluating the effect to continue improving the quality of hospital's project.PDCA cycle method effectively aroused the enthusiasm of young scientific research personnel,and promoted the continuous development of scientific research work.
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OBJECTIVE@#To study the expression of intedeukin-5 (IL-5) mRNA and protein in Nasal polyps and in the inferior turbinate, and to explore the relationship between the expression and the Clinical features.@*METHOD@#Real time fluorescent quantitative PCR(RT-qPCR) and immunohistochemical staining were used to detect the expression of intedeukin-5 (IL-5) mRNA and protein in nasal polyps of 24 cases and in inferior turbinate of 15 cases.@*RESULT@#The expression of IL-5 mRNA in Nasal polyps was 7.52 times higher than that in the inferior turbinate (P 0.01), but was associated with the single or multiple nasal polyps, nasal polyps with or without allergic rhinitis and/or asthma. The differece of expression of IL-5 protein was not associated with the sexual distinction, unilateral or bilateral nasal polyps, primary or recurrent nasal polyps, nasal polyps with or without allergic rhinitis and/or asthma, but was associated with the single and multiple nasal polyps.@*CONCLUSION@#The high expression of IL-5 mRNA and protein is closely related with the formation edema and development of nasal polyps. Some clinical features of Nasal polyps related to the expression level of IL-5 in nasal polyps.
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Female , Humans , Male , Interleukin-5 , Genetics , Metabolism , Nasal Polyps , Metabolism , RNA, Messenger , Genetics , Turbinates , MetabolismABSTRACT
Objective To establish an isolation method for solid GTC in peripheral blood using EpCAM antibody-linked nanobeads and evaluate the sensitivity of the method and its application significance. Methods Five, ten, twenty, fifty and one hundred MCF7 (breast cancer), KYSE70 (esophageal cancer), BxPC-3 (pancreatic cancer) and 9811P (stomach cancer) cells were added into 7. 5 ml erythrocyte lysed peripheral blood obtained from healthy volunteers respectively. EpCAM antibodylinked nanobeads were used to enrich cancer cells. The recovery rates of the in vitro added cancer cells were evaluated by fluorescence microscopy. Then, the untreated thirty cases of esophageal cancer (six cases at stage Ⅰ and Ⅱ, twenty-four cases at stage Ⅲ and Ⅳ), thirty-five cases of breast cancer (fifteen cases at stage Ⅰ and Ⅱ , twenty cases at stage Ⅲ and Ⅳ), thirty cases of pancreatic cancer (five cases at stage Ⅰ and Ⅱ , twenty-five cases at stage Ⅲ and Ⅳ), thirty-three gastric cancer (thirteen cases for stage Ⅰ and Ⅱ ,twenty cases at stage Ⅲ and Ⅳ) were enrolled to enrich the peripheral blood CTC. Thirty healthy volunteers and thirty gastritis patients served as two groups of control. Meanwhile the enriched CTC was identified by IF and HE staining. FISH was used to analyze the copy number of chromosome 8 and chromosome 20 in two hundred esophageal cancer, breast cancer, pancreatic caner and gastric cancer CTC. Results After DAPI staining and mixing with 7.5 ml peripheral blood from healthy donors, the average cell recovery rates of KYSE70, MCF7, BxPC-3 and 9811P cells evaluated under fluorescence microscope were 87%, 87%, 86% and 88% (within group), and the recovery rates of 5 gradient dilution levels were 88%, 85%, 87%, 88% and 87% (intergroup). With a high sensitivity, this method was able to isolate one cancer cell in 107 white blood cells of peripheral blood. The positive rates of more than 2 CTC in the peripheral blood detected by this method were 50% (15/30) of esophageal cancer, 63% (22/35) of breast cancer, 70% (21/30) of pancreatic cancer and 61% (20/33) gastric cancer patients respectively,but no CTC was detected in the peripheral blood of healthy volunteers and gastritis patients (P = 0. 000).The aneusomy of chromosome 8 and chromosome 20 were found in 80% esophageal cancer, 75% breast cancer, 65% pancreatic cancer and 59% gastric cancer. Conclusions The CTC isolation technique with EpCAM antibody-linked nanobeads is sensitive and accurate. The aneusomy of chromosome 8 and 20 is frequent in CTC from esophageal cancer, breast cancer, pancreatic cancer and gastric cancer.
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BACKGROUND: In recent years, nanoparticles has been rapidly developing in tumor hyperthermia, genophore research, and targeted drug therapy, particularly nanoparticle containing drug delivery systems will become another breach in tumor therapy.OBJECTIVE: To summarize the application and mechanism of superparamagnetic nanoparticles for cancer treatment in the medical field.METHODS: A computer-based online search was conducted in Medline for English language publications containing the key words of "superparamagnetic, nanoparticles, targeting" from January 2000 to October 2009. Relevant articles were also searched from CNKI with the same key words in Chinese from January 2005 to October 2009.RESULTS AND CONCLUSION: A total of 123 articles about targeting role of magnetic nanoparticles were included, and there were 24 in Chinese and 108 in English. Articles published earlier, duplicated, and similarly were excluded, and 30 references were finally included. Superparamagnetic nanoparticles characterized by targeting role under external magnetic field, and crystal of ferroso-ferric oxide did not has toxicity to cells. As a gene carrier and drug carrier, superparamagnetic nanoparticles were widely used in medical research and they also provided novel evidences for cancer treatment. By an external magnetic field, how to avoid a comprehensive system of phagocytic endothelial phagocytosis and prevent the course of treatment such as drug-induced thrombus is still inadequate.
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Objective: To study the mechanism of chitosan i n inhibiting the proliferation of rabbit aortic smooth muscle cells(SMCs). Methods: By means of c-myc probe labelled with random primers and Northern blot hybridization, we examined the effect of chitosan on vascu lar SMC c- myc mRNA expression, which was stimulated by newborn bull serum (NB S,20%). Results: The oncogene c-myc mRNA expression incerased in cultured vascular SMC 24 h after NBS exposure. These effects were inhibite d by chitosan (20 μg/ml). Conclusion: Chitosan might inhibit the expression of vascular SMC c-myc mRNA stimulated by NBS, through which the proliferation of vascular SMC are inhibited.