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Article in Chinese | WPRIM | ID: wpr-701218


AIM:To investigate the effect of lutein on the viability of breast cancer cells and its possible mech -anism.METHODS:The human breast cancer T47D cells were divided into control group and lutein(6.25,12.5,25,50 mg/L)treatment groups.The effect of lutein on the viability of T47D cells was measured by MTT assay.The mRNA ex-pression of nuclear factor erythroid 2-related factor 2(Nrf2),glutathione peroxidase 1(GPx1)and superoxide dismutase 2 (SOD2)was detected by RT-qPCR.Fluorescent probes DCFH-DA was used to determine the production of reactive oxygen species(ROS).The protein expression of Nrf2 and p65 was determined by Western blot.RESULTS: The MTT results showed that lutein inhibited T47D breast cancer cell viability in a dose-and time-dependent manner.The RT-qPCR results showed that the mRNA levels of Nrf2, GPx1 and SOD2 were higher in lutein treatment groups than those in the control group(P<0.05),and with the increased concentrations and extension of intervention time of lutein, the relative mRNA levels were all increased.The ROS levels were significantly decreased in the lutein-treated groups(P<0.05).The results of Western blot demonstrated that the protein expression of Nrf 2 was significantly increased(P<0.05), and p65 protein was decreased(P<0.05)in a dose-dependent manner with lutein treatment for 48 h.CONCLUSION: Lutein signifi-cantly inhibits the viability of breast cancer cells,and the inhibition roles may be related to up-regulation of the expression of Nrf2,antioxidant enzymes GPx1 and SOD2 mRNA expression and down-regulation of oxidative stress,thus blocking the NF-κB signaling pathway.

Chinese Journal of Immunology ; (12): 251-255, 2018.
Article in Chinese | WPRIM | ID: wpr-702711


Objective:To investigate the role and mechanism of IL-38 in inhibiting osteoporosis.Methods:A total of 138 cases of patients with osteoporosis in our hospital from June 2014 to December 2016 were recruited.Another 120 cases of fracture surgery patients without osteoporosis were selected as control.Serum levels of IL-38 in different groups were determined using a commercially available sandwich ELISA (Enzyme-Linked ImmunoSorbent Assay).Construction of IL-38-C57BL/6J transgenic mice,the wild type and IL-38 transgenic mice were set to sham operation group (Sham),operation group (ovariectomy,group OVX) respectively.After 8 weeks of the operation,the serum level of alkaline phosphatase(ALP),calcium and phosphorus were detected.The bilateral femur and spine of mice were collected after sacrifice,the morphology and structure of the femur were analyzed,and the bone density was measured by bone density meter.The bone marrow stromal cells(BMSCs) were isolated and the invitro proliferation ability of BMSCs were meas-ured.Western blot were used to detect the phosphorylation level of PI3K,Akt,GSK3β and NFATc1 in BMSCs.After transfection of IL-38 into mouse osteoblast MC3T3-E1 cell,the phosphorylation level of PI3K,Akt,GSK3β and NFATc1 were detected by Western blot.Apoptosis of MC3T3-E1 cells were detected by flow cytometry.Results:The serum level of IL-38 in patients with osteoporosis were significant lower than control group(P<0.05).The serum level of estrogen,calcium and phosphorus in OVX group of wild type and IL-38 transgenic mice were significant lower than the sham operation group(P<0.05),while the level of ALP was significant higher than sham operation group (P<0.05),but the serum level of calcium and phosphorus in OVX group of IL-38 transgenic mice were significant higher than wild type mice(P<0.05).The pathological section of femur and spine BMD showed that the bone tissue in wild type mice and IL-38 transgenic mice in OVX group were damaged and the bone density decreased significantly,but IL-38 transgenic mice was significant better than wild type mice (P<0.05).The proliferation ability of BMSCs in OVX group of IL-38 transgenic mice was significant higher than wild type mice (P<0.05).The phosphorylation level of PI3K,Akt and NFATc1 in OVX group of IL-38 transgenic mice were significant lower than wild type mice(P<0.05),while the phosphorylation level of GSK3β was significant higher than wild type mice (P<0.05).After transfection of IL-38 into MC3T3-E1 cell,the phosphorylation level of PI3K,Akt and NFATc1 were significant decreased (P<0.05),while the phosphorylation level of GSK3β was significant increased (P<0.05).Flow cytometry assay showed that IL-38 transfection significant decreased the apoptosis of MC3T3-E1 cells(P<0.05).Conclusion:The serum level of IL-38 in patients with osteoporosis is decreased significantly.IL-38 may inhibit the proliferation of BMSCs and inhibit the apoptosis of osteoblasts by regulating the PI3K/Akt/GSK3β/NFATc1 signaling pathway.

Article in Chinese | WPRIM | ID: wpr-664767


Objective:To assess the midterm clinical and radiological outcomes of internal fixation and fusion for the treatment of Hirayama disease and to evaluate the clinical significance and value of this procedure.Methods:In the study,36 patients were treated with anterior cervical internal fixation and fusion.The clinical outcomes including muscle strength and atrophy were recorded.The radiological outcomes including range of motion of cervical spine and the cross-sectional area of spinal cord at each level on MRI scan were measured before and at 3 month,1 year and 2 years follow-up time points after surgery.Results:(1) Clinical outcomes:all the patients showed no further progression of symptoms except one patient with mild progression of muscular weakness and atrophy.As the time passed by,the ratio of the patients with muscle strength and atrophy improvement increased.There were 26.5 % of patients in 3 months,36.0% in 1 year and 85.7% in 2 years who experienced muscle strength improvement.8.8% of patients in 3 months,24.0% in 1 year and 35.8% in 2 years felt muscle atrophy improvement.And 12 of the 14 patients showed improved muscle strength and atrophy at the end of 2 years period follow-up.(2) Radiological outcomes:the range of motion (ROM) of C2-C7 was significantly decreased after the operation.The ROM of preoperation was 62.25° ±2.10° and that of 2 years postoperation was 13.67° ± 7.51°(P < 0.01).The spinal cord was of no compression on flexion MRI.The cross-section area of spinal cord on MRI was significantly increased only at C6 level (P <0.05) at the end of three months follow-up.The level of increased cross-section area rose to C4-C5-C6 levels (P <0.01) in 1 year and to C4-C5-C6-C7 levels at the end of 2 years follow-up P < 0.05).The cross-section area increased 15.60% at C4,19.08% at C5,21.60% at C6 and 23.91% at C7 with significant difference (P <0.05) 2 years after the operation.Conclusion:Anterior cervical internal fixation and fusion is an effective surgical treatment for Hirayama disease and may provide preferable midterm clinical and radiological outcomes.This procedure has clinical significance and value in terms of control of the progression and outcome of this disease.

Article in Chinese | WPRIM | ID: wpr-733229


Objective To investigate the effect of ursolic acid (UA) on retinal vascular form in oxygen-induced mouse retinopathy.Methods The 60 clean 7-day C57BL/6J mice were divided into 6 groups randomly:blank control group,model control group (PBS),positive control group (triamcinolone),UA intervention group (low dose,meta dose,high dose),with 10 mice in each group(right eye as experimental subject,10 eyes each group).The blank control group mice were raised in air,with other groups of mice in (750 ± 20) mL/L O2 high-oxygen environment for 5 consecutive days.The model control group mice and breastfeeding mice were put back in air enviroment(210 mL/L O2) on the 12th day after birth to induce the generation of retinal neovascufarization.The drug treatment was applied to the corresponding groups immediately when models were successful:3 μL sterile PBS was injected in model control group,3 μL 1.5,3.0 and 6.0 μg UA in UA intervention group,3 μL triamcinolone(1 mL ∶ 40 mg) in positive control group.All mice were killed after overdose of anesthesia on the 17th day,and their eyeballs were made into retinal tissue sections,HE dying,counting the neovasculanzation endothelial nuclei number which broke the retinal internal limiting membrane.The morphologic changes of retinal vessels were estimated by observing the vascular pattern with the technology of stretched preparation of retina.Results From the observation of tissue pathology slice in blank control group,the structure of internal limiting membrane was even,and the vascular endothelial cells lined up evenly,while some vascular endothelial cells broke the internal limiting membrane occasionally.In model control group,a lot of vascular endothelial cells broke the internal limiting membrane and new vessel lumen was formed.The result of low UA intervention group was basically similar to that of model control group.The result of high-dose UA intervention group was close to that of positive control group,though some vascular endothelial cells broke the internal limiting membrane,and internal limiting membrane structure was even,and new vessel lumen was not formed.The endothelial nuclei number of newly-generated internal limiting membrane vessel in model control group was obviously higher than that of blank control group,while the high-dose UA intervention group was obviously lower than that of model control group,and the high-dose UA intervention group was lower than that of the low-dose UA intervention group obviously,and all the differences were statistically significant(P < 0.05).Retinal flatmounts showed that in the blank control group retinal vascular appeared with uniformly radiated distribution from optic disc in all directions,with bigger pipe diameter,proper branch and clear peripheral retinal vascular structure.However,lots of retinal neovascularization was seen in model control group,with slender retinal diameter,line rigidity,structure disorder,and uneven distribution,and there was large non-perfused areas in the centeral area.Through comparison with different doses UA intervention group,retinal vascular distribution and pattern in the high-dose UA intervention group were close to the positive group.The distribution of retinal neovascularization was much better than the model control group and the low-dose UA intervention group,with no obvious non-perfused areas.Conclusions UA can inhibit the formation of neovascularization in oxygen-induced mice retinal ischemia model.It has a positively correlated relationship with UA dose.