1.Recognition of environmental estrogen-like compounds based on fluorescent imaging analysis of ERα-EGFP nuclear granules formation
Military Medical Sciences 2015;(1):12-17
Objective To establish a quick method to probe the environmental estrogen-like compounds via fluorescent imaging analysis in high content analysis (HCA) technology.Methods HCA assays were performed to quantitatively in-vestigate the effect of collected environmental pollutants , including bisphenol A , nonylphenol , 1,2-phenylenediamine , 4-aminophenol, resorcinol, 3-aminophenol, 1,4-phenylenediamine and 2,5-diaminotoluene on nuclear granules formation ( Foci-formation) of estrogen receptor α( ERα)-EGFR( enhanced green fluorescent protein ) fusion-protein, which was se-lected as a research model in this study .The results were confirmed by the ERαtranscriptional activity by luciferase as-says.Results Compounds 1,2-phenylenediamine, 4-aminophenol, resorcinol, 3-aminophenol, 1,4-phenylenediamine and 2,5-diaminotoluene sulfate could not induce the ERα-EGFR nuclear granule formation.17-β-Estradiol, bisphenol A, or nonylphenol enhanced ERα-EGFR nuclear granule formation in a dose-dependent manner .The EC50 value was (4.17 ± 0.41) nmol/L, (1.48 ±1.79) μmol/L,or (3.70 ±0.78) μmol/L, respectively.The minimum detectable concentration was 1 nmol/L (17-β-estradiol)and 300 nmol/L (bisphenol A, nonylphenol).In luciferase tests, 17-β-estradiol, bisphe-nol A, or nonylphenol increased ERαtranscriptional activity in a dose-dependent manner ,and the EC50 value was (4.46 ±0.56) nmol/L, (2.31 ±0.21) μmol/L, or (6.60 ±0.94) μmol/L, respectively.The minimum detectable concentration was 3 nmol/L (17-β-estradiol), 300 nmol/L (bisphenol A),and 1 μmol/L (nonylphenol).Conclusion As an efficient method for ERαagonist identification , HCA assays based on the cell image phenotypes analysis can be used in quick recog -nition of environmental compounds with ER agonist-like activity.In all experimental compounds , only bisphenol A and non-ylphenol have a clear ER agonist-like activity .
2.One case of granular cell tumor of the breast
Fan FENG ; Tielin WANG ; Guosheng REN ; Fan LI
Chinese Journal of Endocrine Surgery 2017;11(2):161-163
3.Effect of adipose-derived mesenchymal stem cell transplantation on inflammatory response and ventricular remodeling after myocardial infarction
Yan FAN ; Jianjun WANG ; Feng WEI ; Xiaohai FAN ; Aiqun MA
Chinese Journal of Tissue Engineering Research 2014;(6):900-905
BACKGROUND:Whether adipose-derived mesenchymal stem cells are able to exert immunomodulatory effects in the treatment of myocardial infarction, as wel as the best time, is less reported.
OBJECTIVE:To observe the effect of adipose-derived mesenchymal stem cells on inflammatory reaction and ventricular remodeling after myocardial infarction, and to explore the possible mechanisms of adipose-derived mesenchymal stem cells for the treatment of myocardial infarction.
METHODS:Enzyme digestion method was employed to isolate and culture rat adipose-derived mesenchymal stem cells. By ligation of the left anterior descending coronary artery, we established animal models of myocardial infarction in 40 rats. The rats were randomly divided into four groups:sham group, control group (injected high-glucose Dulbecco’s modified Eagle’s medium), 3-hour transplantation group (transplanted adipose-derived mesenchymal stem cells after 3 hours of myocardial infarction), 7-day transplantation group (transplanted adipose-derived mesenchymal stem cells after 7 days of myocardial infarction). After 14 days of operation, the levels of tumor necrosis factor-αand interleukin-10 in the plasma were detected by enzyme linked immunosorbent assay. After 28 days of operation, the left ventricular end diastolic diameter, left ventricular end systolic diameter, left ventricular ejection fraction and left ventricular fractional shortening were measured by echocardiography.
RESULTS AND CONCLUSION:Compared with the control group, in the 3-hour transplantation group and 7-day transplantation group, the levels of tumor necrosis factor-αwere significantly lower (P<0.01), and the levels of interleukin-10 were significantly higher (P<0.01) at postoperative 14 days;the left ventricular end diastolic diameter and left ventricular end systolic diameter in the two transplantation groups were also significantly smal er (P<0.05), but left ventricular ejection fraction and left ventricular fractional shortening were significantly elevated (P<0.05), which was more apparent in the 3-hour transplantation group than the 7-day transplantation group. Adipose-derived mesenchymal stem cells transplantation in acute phase of myocardial infarction can suppress the inflammatory response, regulate the cytokine network equilibrium, and thus delay ventricular remodeling and improve cardiac function.
4.Research progress of polyethylene inserts wear measurement and evaluation in total knee arthroplasty.
Feng ZHAO ; Chuan WANG ; Yubo FAN
Chinese Journal of Medical Instrumentation 2015;39(1):33-36
Wear of polyethylene (PE) tibial inserts is a significant cause of implant failure of total knee arthroplasty (TKA). PE inserts wear measurement and evaluation is the key in TKA researches. There are many methods to measure insert wear. Qualitative methods such as observation are used to determine the wear and its type. Quantitative methods such as gravimetric analysis, coordinate measuring machines (CMM) and micro-computed tomography (micro-CT) are used to measure the mass, volume and geometry of wear. In this paper, the principle, characteristics and research progress of main insert wear evaluation method were introduced and the problems and disadvantages were analyzed.
Arthroplasty, Replacement, Knee
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Humans
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Knee Prosthesis
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Polyethylene
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Prosthesis Design
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Prosthesis Failure
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Tibia
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Tomography, X-Ray Computed
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X-Ray Microtomography
5.Clinical Curative Effect of Xintong Decoction on Patients with Unstable Angina Pectoris and Impact on Serum Matrix Metalloproteinase 2
Jinru FAN ; Man WANG ; Feng LIU
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(05):-
Objective To observe the clinical curative effect of Xintong decoction in treating unstable angina pectoris(UAP) and impact on serum matrix metalloproteinase 2(MMP-2) of patients with syndrome of phlegm and blood stasis,qi stagnation and obstruction of heart meridian,and probes into its intervention mechanism on UAP.Method Patients were randomly divided into treatment group(30 cases,treated with Xiaotong decoction + routine western medicine) and control group(30 cases,treated with routine western medicine).The course was four weeks.The clinical curative effect was observed and serum MMP-2 before and after treatment was detected.Results TCM syndrome score,angina pectoris,nitroglycerin discontinue rate,serum concentration of MMP-2 in the treatment group were significantly better than the control group.Conclusion Xintong decoction has good curative effect in treating UAP.It can reduce the serum level of MMP-2 of patients with UAP,so as to slowing the progress of plaque and stabilizing plaques.
6.Expressions of TFPI-2 and MMP-9 in esophageal squamous cell carcinoma tissue and their relationship with vasculogenic mimicry
Luchuan ZHANG ; Feng WANG ; Qingxia FAN
Journal of Jilin University(Medicine Edition) 2014;(3):664-667
Objective To investigate the expressions of tissue factor pathway inhibitor-2 (TFPI-2 ) and matrix metalloproteinase-9 (MMP-9)in esophageal squamous cell carcinoma(ESCC)tissue and their relationship with vasculogenic mimicry (VM).Methods 162 cases of esophageal squamous cell carcinoma tissues were collected. CD34/periodic acid-schiff double staining was performed to observe the distribution of VM in ESCC tissue,and immunohistochemical staining was used to detect the expressions of TFPI-2 and MMP-9 in ESCC tissue. The relationship between VM and the clinicopathologic parameters of ESCC, the expressions of TFPI-2 and MMP-9 were analyzed.Results The positive rate of VM in ESCC tissue was 20.37%.The positive rate of VM in poorly differentiated group(40.38%)was significantly higher than those in moderately differentiated group(11.76%)and well differentiated group (7.14%)(χ2 = 20.915, P < 0.01 ). The positive rate of VM in TNM Ⅰ-Ⅱ(11.59%)group was significantly lower than that in TNM Ⅲ group(26.88%)(χ2=5.707,P=0.017).The positive rate of TFPI-2 in VM(+)group(33.34%)was significantly higher than that in VM(-)group(6.45%) (χ2=4.582,P=0.032)in poorly differentiated ESCC.The positive rate of MMP-9 in VM(+)group(78.79%) was significantly higher than that in VM(-)group(44.96%)(χ2=12.05,P=0.001).The expression level of TFPI-2 in poorly differentiated group was positively correlated with VM (r= 0.166,P= 0.032 ), and the expression level of MMP-9 was positively correlated with the VM(r=0.183,P=0.018).The five-year survival rate in VM (-) group was significant higher than that in VM (+) group (χ2 = 22.84, P < 0.001 ). Conclusion VM exists in ESCC tissue,especially in poorly differentiated and advanced ESCC tissue.VM is related to poor prognostis of ESCC.TFPI-2 and MMP-9 might involve in the formation of VM in ESCC.
7.Research on heart function in pregnant women with sinus tachycardia evaluated with echocardiography
Lingyun FAN ; Yanru WANG ; Xiaoli FENG
Chinese Journal of Postgraduates of Medicine 2014;37(9):16-18
Objective To investigate the characteristic and clinical value of echocardiography in pregnant women with sinus tachycardia.Methods Thirty pregnant women with sinus tachycardia (experiment group) and 30 healthy pregnant women (control group) were selected.The echocardiography results and clinical data were compared between the 2 groups.Results There were no statistical differences in age,gestational age,blood pressure,weight between the 2 groups (P > 0.05).The heart rate in experiment group was significantly higher than that in control group [(123.20 ± 13.23) times/min vs.(86.17 ± 6.78) times/min],there was statistical difference (P < 0.01).The left ventricular posterior wall thickness and the rates of mitral regurgitation,tricuspid regurgitation,aortic regurgitation in experiment group were significantly higher than those in control group [(10.23 ± 1.30) mm vs.(8.79 ± 1.90) mm,63.33% (19/30) vs.16.67% (5/30),66.67%(20/30) vs.30.00%(9/30),26.67%(8/30) vs.6.67%(2/30)],the left ventricular posterior wall motion amplitude was significantly lower than that in control group [(8.07 ± 1.00) mm vs.(9.26 ± 1.71) mm],there were statistical differences (P < 0.01 or < 0.05).There were statistical differences in the indexes of left ventricular systolic function and diastolic function between the 2 groups (P <0.01 or <0.05).Conclusion The anatomical indexes and function indexes of echocardiography in pregnant women with sinus tachycardia are used to change,and the echocardiography in time can provide the basis of treatment.
8.Overexpression of connexin 43 in bone marrow mesenchymal stem cells in Xiao Meishan swines.
Yixuan FAN ; Yanli ZHANG ; Qiang WANG ; Caifang REN ; Feng WANG
Chinese Journal of Biotechnology 2015;31(3):351-360
We studied the function of connexin 43 (Cx43) gene in Xiao Meishan swine bone marrow mesenchymal stem cells (BMSCs) resulting from overexpression. Cx43 eukaryotic expression vector (pEGFP-Cx43) was constructed and transfected into BMSCs by nucleofector, after detecting the transfection efficiency; the expression of Cx43 was verified by RT-PCR, immunofluorescence and western blotting. Furthermore, we detected its cell cycle and apoptosis through flow cytometry. Our results show that pEGFP-Cx43 plasmid was successfully constructed, and green fluorescence in pEGFP-Cx43 transfected BMSCs was highly expressed with 60% transfection efficiency. In transgenic Xiao Meishan swines BMSCs, the expression level of Cx43 mRNA and protein were up-regulated. Meanwhile, the ability of cell proliferation was significantly increased, and the apoptosis rate was significantly reduced. Taken together, Cx43 overexpression could promote the proliferation of Xiao Meishan swine's BMSCs and markedly reduce their apoptosis, which provides evidence for in vivo research.
Animals
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Animals, Genetically Modified
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Apoptosis
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Cell Proliferation
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Connexin 43
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metabolism
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Flow Cytometry
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Genetic Vectors
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Mesenchymal Stromal Cells
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metabolism
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Plasmids
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Swine
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genetics
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Transfection
9.Three-dimensional Culture of Chondrocyte Using Methacrylic Alginate Gel Beads Cross-linked with Mixed Metal-cation.
Yang WANG ; Yuxia FENG ; Xing FAN ; Liling REN
Journal of Biomedical Engineering 2015;32(3):599-604
This study was to explore a better three-dimensional (3-D) culture method of chondrocyte. The interpenetrating network (IPN) gel beads were developed through a photo-cross linking reaction with mixed barium ions and calcium ions at the ratio of 5:5 with the methacrylic alginate (MA), which was a chemically conjugated alginate with methacrylic groups. The second generation of primary cartilage cells was encapsulated in the MA gel beads for three weeks. In the designated timing, HE stain, Alamar blue method and Scanning electron microscopic were used to determine the cartilage cells growth, proliferation and the cell distribution in the scaffolds, respectively. The expression of type II collagen was investigated by an immunohistochemistry assay and the glycosaminoglycan content was quantitatively evaluated with the spectrophotometry of 1, 9 dimethylene blue assay. Compared to the alginate control group, the deposition of glycosaminoglycan was significantly upregulated in IPN-MA gel beads with higher cell proliferation. The secretion of extracellular matrix and proliferation of chondrocyte in methacrylic alginate gel beads were higher than that in Alginate beads. Cells were able to attach, to grow well on the scaffolds under scanning electron microscopy. The result of immunohistochemistry staining of collagen type II was positive, confirming the maintenance of chondrocyte phenotype in methacrylic alginate gel beads. This study shows a great potential for three-dimensional culture of cartilage.
Alginates
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chemistry
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Barium
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chemistry
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Calcium
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chemistry
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Cartilage
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cytology
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Cations
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Cell Culture Techniques
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instrumentation
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Cells, Cultured
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Chondrocytes
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cytology
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Collagen Type II
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chemistry
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Glucuronic Acid
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chemistry
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Glycosaminoglycans
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chemistry
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Hexuronic Acids
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chemistry
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Metals
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chemistry
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Microscopy, Electron, Scanning
10.Two-stage exchange total hip arthroplasty for infected hip surgery
Zhefeng CHEN ; Qing WANG ; Weiding CUI ; Weimin FAN ; Feng LIU
Chinese Journal of Orthopaedics 2012;32(9):817-822
Objective To evaluate effect of two-stage exchange total hip arthroplasty (THA) using an antibiotic-loaded cement spacer in the treatment of hip infection secondary to hip surgery.Methods From January 2005 to January 2010,6 consecutive patients with infected hip secondary to hip surgery,including 2 males and 4 females,aged from 43 to 68 years (average,59.7±9.2 years) were treated with two-stage exchange THA.There were 3 cases of femoral neck fracture treated with compression screws fixation,1 case of femoral head necrosis treated with bone graft,and 2 cases of early stage femoral head necrosis treated with core decompression and local interventional therapy respectively.Debridement and insertion of antibioticloaded cement spacer was performed in the first stage.After eradication of infection,it was converted to THA in the second stage.All patients were followed up regularly.Erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) were tested.Harris score was used to evaluate the function of the hips.Results All patients were followed up for an average of 46 months (range,24 to 81 mouths).Five of 6 patients were successfully converted to THA after an average of 14 weeks.One patient accepted the second debridement and reinsertion of a spacer owing to the abnormal values of ESR and CRP; this case was converted to THA successfully 12 weeks after the second debridement.Harris hip score improved from preoperative 35.6±3.3 to 57.8±5.4 between the two stages and 92.3±5.7 at final follow-up.One patient suffered deep vein thrombosis in the infected extremity and pulmonary embolism.There was no reinfection after THA.Conclusion Twostage exchange THA using an antibiotic-loaded cement spacer can obtain satisfactory results for the treatment of hip infection secondary to the hip surgery.