1.Wnt3a-induced extracellular matrix production and gel contraction in human lens epithelial cells
Xiuli, BAO ; Tingting, LIU ; Yanyan, WANG ; Lin, LI
Chinese Journal of Experimental Ophthalmology 2016;34(7):597-601
Background Excessive production,deposition and contraction of extracellular matrix (ECM) of human lens endothelial cells (LECs) is one of main causes to posterior capsular opacification (PCO).Researches indicated that Wnt3a protein was involved in production of ECM and fibrosis in epithelial cells,but its effect on LECs is still unclear.Objective This study aimed to elucidate the roles of Wnt3a in production of ECM and gel contraction of LECs.Methods Lipofectamine-mediated transient transfection technique was used to introduce cDNA of Wnt3a gene and pcDNA3-HA vector into the LEC cell line SRA01/04 to act as Wnt3a transfection group and control group respectively.After 48 h of transfection,Western blot was used to detect the expression of Wnt3a,main components of ECM Col-Ⅰ,Col-Ⅳ and integrin β1.Immunofluorescence was used to detect the expression and distrubition of α-SMA and F-actin;collagen contraction was observed by mingling SRA01/04 cells with Col-Ⅰ.Results After 48 hours of transfection using lipofectamine 2000,the expressions of wnt3a protein in SRA01/04 cells was 0.703 ±0.105 in the wnt3a transfected group,and that in the control group was 0.290 ± 0.066,showing a significant difference between the two groups (t =5.782,P<0.01).Western blot assay showed that the expression levels of Col-Ⅰ and Integrin β1 was 0.697±0.021 and 0.875±0.055 in the Wnt3a transfected group,and which was significantly higher than 0.370±0.020 and 0.580±0.030 in the control group (t =19.600,8.156,both P<0.01).The expression level of Col Ⅳ in the Wnt3a transfected group was higher than that of the control group (0.430±0.020 vs 0.383 ±0.031),but the difference was not significant (t =2.514,P>0.05).Immunofluorescence assay revealed that F-actin and α-SMA were weakly expressed in the cell membrane primarily in the control group,while they were strongly expressed in the cell membrane,cytoplasma in the Wnt3a transfected group.Tewnty-four hours after addtion of Col-Ⅰ,the gel contraction area ratio appeared to be more obvious in comparison with the 8 hours (64.1% ±2.3% vs 98.9% ± 1.0%),and gel contraction area ratio was lower in the Wnt3a transfected group than that in the control group (64.1% ± 2.3% vs 93.9% ± 3.1%).Conclusions The overexpression of Wnt3a activates the production of ECM,and the remodeling of celluar skeleton and cellular contraction.
2.Finite element analysis of total hip replacement.
Xiuli DU ; Lan CHEN ; Genlin XU ; Yiwang BAO
Journal of Biomedical Engineering 2009;26(2):429-432
Total hip replacement is a very effective method to cure many kinds of hip joint illnesses. About a century ago, it was first used in clinic. Since then, total hip replacement has been well developed. Hip joints sustain the most load of body, so people pay great attention to the hip prothesis' mechanics property. Especially after the finite element analysis was widely used in biomechanics investigation, the stress distribution of different designs can be easily compared with each other, and the relatively better parameters of the design could be decided. The stress distribution of different materials with the same design also can be valued. However, studies have indicated that total hip joint replacement still has some disadvantages. Loosening of the hip prothesis is still the most likely cause of the failure of surgery, and generally this is believed to stem from either mechanical failure of the fixation in response to over high density stresses, or osteolysis of the surrounding bone stock responsing to particular wear debris. Many researchers on computational studies have considered the potential for the former one, but only a few have attempted to tackle the latter. The process of osteolysis of the bone is not yet completely known. Nowadays, in order to solve the problems of loosening, investigators are trying to find different methods. Some of them are working on improving the geometry parameters and the shape of the hip prothesises, some are trying to find new suitable biomaterials, and, at the same time, the fixation methods are under deliberation.
Arthroplasty, Replacement, Hip
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adverse effects
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Computer-Aided Design
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Finite Element Analysis
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Humans
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Prosthesis Design
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Prosthesis Failure
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Stress, Mechanical
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Weight-Bearing
3.Inhibitory effect and biological mechanism of Dickkopf-1 on the proliferation of human lens epithelial cells
Chinese Journal of Experimental Ophthalmology 2020;38(4):285-290
Objective:To investigate the effects of Dickkopf-1(DKK1) on the proliferation of human lens epithelial cells (LECs) and its possible mechanism in order to search a new target for the treatment of posterior capsular opacification (PCO).Methods:Human LECs line (SRA 01/04 cells )were divided into Wnt3a overexpression group, DKK1 group and control group.Wnt3a gene expression vector was transfected into SRA 01/04 cells by liposome mediated transfection to establish a PCO model in the Wnt3a overexpression group, DKK1 of 100 μg/ml was added into the medium 48 hours after transfection of Wnt3a gene vector in the DKK1 group, and only pcDNA3-HA vector was transfected in the control group.The survival rate of SRA 01/04 cells was detected with a cell counting kit-8 (CCK-8) assay.The expression rate of proliferating cell nuclear antigen (PCNA) in the cells was detecteds by immunocytochemistry.The expression of β-catenin in the cells was detected and located by immunofluorescence.The expression of Wnt3a CyclinD1 and C-Myc were detected by Western blot assay.Results:The relative expression of Wnt3a protein in the control group was 0.49±0.07, which was significantly lower than that in the Wnt3a overexpression group (0.84±0.06) ( t=3.704, P=0.02). The survival rate in the Wnt3a overexpression group, DKK1 group and control group showed significant difference over time ( Fgroup=10.910, P<0.05; Ftime=6.041, P<0.05). The survival rate in the Wnt3a overexpression group was significantly increased in comparison with the control group and that in the DKK1 group was significantly reduced in comparison with the Wnt3a overexpression group(all at P<0.05). β-Catenin was expressed mainly in cytoplasm and cell nucleus in the Wnt3a overexpression group and only in cytoplasm in the DKK1 group. in the control group, β-catenin showed a weaked expression in the cytoplasm and nucleus in comparison with the Wnt3a overexpression group.The expression rates of PCNA protein were (9.4±1.4)%, (43.4±5.4)%, and (14.2±2.3)% in the control group, Wnt3a overexpression group and DKK1 group, respectively, with a significant difference among the groups ( F=28.250, P<0.05), and the expression rates of PCNA protein were significantly reduced in the control group and DKK1 group compared with the Wnt3a overexpression group (both at P<0.05). β-Catenin protein were expressed mainly in the cytoplasm and nucleus in the Wnt3a overexpression group and only in the cytoplasm in the control group.In the DKK1 group, the expression of β-catenin protein was weakened in the cytoplasm and nucleus in comparison with the Wnt3a overexpression group.The relative expressions of CyclinD1 were 0.64±0.07、0.84±0.03 and 0.55±0.10, C-Myc were 0.59±0.05、0.93±0.02 and 0.47±0.08 in the control group, Wnt3a overexpression group and the DKK1 group, respectively with significant differences among the groups ( F=20.580, 5.040, both at P<0.05). The relative expressions of CyclinD1 and C-Myc in the Wnt3a overexpression group were significantly higher than those in the DKK1 group and the control group (all at P<0.05). Conclusions:DKK1 inhibits activation of Wnt/β-catenin signaling pathway induced by Wnt3a overexpression in SRA01/04 cells, and down-regulation of downstream target proteins cyclin D1 and C-Mgc may be the biological mechanism of Dkk1 inhibiting human LECs proliferation.
4.Perioperative nursing for patients with osteoporotic vertebral compression fractures receiving vertebroplasty by using high viscosity bone cement
Tingye HU ; Yuhe LU ; Weifu Lü ; Yonghui ZHANG ; Ling XIN ; Kai WANG ; Lei XIA ; Jinhua ZHENG ; Huiling CAO ; Xiaoxing ZHOU ; Xiuli BAO
Journal of Interventional Radiology 2017;26(3):274-276
Objective To discuss the perioperative nursing measures for patients with osteoporotic vertebral compression fractures who are receiving percutaneous vertebroplasty (PVP) treatment by using high viscosity bone cement.Methods A total of 30 patients with osteoporotic vertebral compression fractures were included in this study.All patients were treated with PVP by using high viscosity bone cement.Preoperative routine nursing,psychological intervention,dietary intervention,postoperative guidance of rehabilitation exercise of limbs,close observation of bone cement leakage were strictly implemented,and the corresponding nursing measures were promptly taken when needed.Results Through strict implement of the nursing intervention all 30 patients could actively cooperate with PVP treatment,and after PVP the pain was significantly relieved in all patients.Conclusion Adequate preoperative preparation,proper postoperative guidance,careful observation and effective nursing can help the patients resume their daily life activities as soon as possible,relieve the pain,and improve the quality of life as well.(J Intervent Radiol,2017,26:274-276)
5.Carvacrol pretreatment attenuates myocardial oxidative stress and apoptosis following myocardial ischemia-reperfusion in mice.
Xudong SONG ; Aihua CHEN ; Yingfeng LIU ; Xian-Bao WANG ; Yijun ZHOU ; Lei LIU ; Xiuli ZHANG ; Lizi WANG ; Pingzhen YANG
Journal of Southern Medical University 2013;33(11):1624-1627
OBJECTIVETo investigate the effect of carvacrol pretreatment on myocardial ischemia-reperfusion (I/R) injury and its underlying mechanisms.
METHODSWild-type male C57 BL/6 mice were randomized into 5 groups (n=13), namely the sham-operated group, vehicle (DMSO in saline)+ I/R group, carvacrol (20 mg/kg) + I/R group, carvacrol (40 mg/kg) + I/R group, and carvacrol (60 mg/kg) + I/R group. The mouse models of myocardial I/R injury were established by a 45-min occlusion of the left anterior descending coronary artery (LAD) followed by reperfusion for 2 h. Carvacrol or vehicle was administered intravenously 15 min before LAD occlusion. After reperfusion, the mice were examined for myocardial oxidative stress level and apoptosis rate.
RESULTSCompared with the vehicle group, the 3 carvacrol-pretreated groups showed significantly reduced myocardial infarct size, oxidative stress level and cardiac myocyte apoptosis rate (P<0.01).
CONCLUSIONCarvacrol can protect against myocardial I/R injury by inhibiting myocardial oxidative stress and apoptosis in mice.
Animals ; Antioxidants ; pharmacology ; Apoptosis ; drug effects ; Cardiotonic Agents ; pharmacology ; Male ; Mice ; Mice, Inbred C57BL ; Monoterpenes ; pharmacology ; Myocardial Infarction ; pathology ; Myocardial Reperfusion Injury ; metabolism ; pathology ; Myocytes, Cardiac ; cytology ; drug effects ; Oxidative Stress ; drug effects ; Random Allocation