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Objective:To investigate the effect of epigallocatechin gallate (EGCG) on the activation of human Tenon fibroblasts (HTFs) and its mechanism.Methods:Tenon capsule tissues from nine eyes of nine advanced primary open angle glaucoma patients during trabeculectomy were obtained for primary cell culture.HTFs harvested were identified by immunofluorescence staining for vimentin and keratin.Cells at passage 4-6 were used for experiment.Viability of HTFs treated with EGCG at 0, 10, 20, 30, 40, 50, 60, 70 and 80 μmol/L was detected by cell counting kit-8 (CCK-8) assay.The cells were divided into blank control group, transforming growth factor (TGF)-β1-induced group, and EGCG-treated group, which were cultured in normal medium, medium containing 10 ng/ml TGF-β, medium containing 10 ng/ml TGF-β+ 50 μmol/L EGCG, respectively.The proliferation rate of HTFs was detected by BrdU labeling assay.Cell migration was observed by scratch wound healing assay.The expression of α-smooth muscle actin (α-SMA) was measured by immunofluorescence staining.The protein relative expression levels of Smad2/3, phosphoinositide-3-kinase (PI3K), protein kinase B (Akt) as well as the phosphorylated Smad2/3 (p-Smad2/3) and phosphorylated Akt (p-Akt) were measured by western blot.This study was approved by the Ethics Committee of Guangdong Provincial People's Hospital (NO.GDREC2019331H[R1]).Results:The HTFs harvested had spindle shape, grew regularly and were vimentin-positive.CCK-8 assay showed that there was no significant difference in the variability of HTFs treated with EGCG at 10, 20, 30, 40 and 50 μmol/L in comparison with 0 μmol/L EGCG treatment (all at P<0.05). BrdU labeling assay showed that cell proliferation in the TGF-β1-induced group was (66.37±12.65)%, which was significantly higher than (14.75±12.33)% in EGCG-treated group ( P<0.05). Three days after scratch, the relative scratch area in the TGF-β1-induced group was (47.33±12.22)%, which was significantly lower than (92.67±4.04)% in the EGCG-treated group ( P<0.05). Immunofluorescence assay showed that α-SMA fluorescence was significantly enhanced in the TGF-β1-induced group in comparison with the blank control group, which was reduced to blank control group level in EGCG-treated group.Western blot analysis showed that there were significant differences in the relative expression levels of p-Smad2/3, PI3K and p-Akt protein among the various groups ( F=58.820, 121.153, 69.289; all at P<0.001). The relative expressions of p-Smad2/3, PI3K and p-Akt in the TGF-β1-induced group were significantly higher than those in the blank control group, 10 μmol/L and 50 μmol/L EGCG-treated groups (all at P<0.05). Conclusions:EGCG can suppress TGF-β1-induced HTFs activation through Smad and PI3K/Akt signaling pathways.
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Allergen testing for the diagnosis and treatment of allergic diseases is essential , with the development of detection technology , automatic quantitative detection technology has become an important means of allergen detection and development trend .Compared with qualitative test , quantitative test can be more accurate in the diagnosis of allergic diseases , and can more effectively reflect the incidence of allergy , predict the occurrence and prognosis of the disease , and monitor the treatment effect.Therefore, although there are still many challenges in the quantitative detection of allergens , the automatic quantification of allergen detection is still an inevitable trend .
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The changes in the levels of serum interleukin-13 (IL-13) and nerve growth factor (NGF) in patients with systemic lupus erythematosus (SLE) and their clinical significance were investigated. Sandwich ELISA was used to determine the levels of serum IL-13 and NGF in 35 SLE patients and 15 normal controls. The results showed that the levels of serum IL-13 (92.69+/-9.87 pg/ml) and NGF (339.69+/-25.60 pg/ml) in active SLE patients were significantly higher than those in inactive SLE patients (IL-13, 54.22+/-9.31 pg/ml; NGF, 300.89+/-33.51 pg/ml) (P<0.01). The inactive patients also had significantly increased serum levels of IL-13 and NGF as compared with normal controls (IL-13, 35.20+/-12.70 pg/ml; NGF, 111.40+/-32.54 pg/ml; P<0.05 and P<0.01, respectively). Spearman correlation analysis revealed that the serum IL-13 levels were correlated with disease activity index of SLE (SLEDAI), ESR and serum levels of C3 (r= 0. 813, 0.504, -0.605, respectively). The serum NGF levels were also correlated with above markers (r=0.442, 0.338, -0.463, respectively). The serum levels of IL-13 and NGF had a positive correlation (r=0.506, P<0.01). It was suggested that IL-13 and NGF might be involved in the pathogenesis of SLE and closely correlated with disease activity.
Subject(s)
Enzyme-Linked Immunosorbent Assay/methods , Interleukin-13/blood , Lupus Erythematosus, Systemic/blood , Lupus Erythematosus, Systemic/etiology , Nerve Growth Factors/bloodABSTRACT
Summary: The changes in the levels of serum interleukin-13 (IL-13) and nerve growth factor (NGF) in patients with systemic lupus erythematosus (SLE) and their clinical significance were investigated. Sandwich ELISA was used to determine the levels of serum IL-13 and NGF in 35 SLE patients and 15 normal controls. The results showed that the levels of serum IL-13 (92.69±9.87 pg/ml) and NGF (339.69±25.60 pg/ml) in active SLE patients were significantly higher than those in inactive SLE patients (IL-13, 54.22±9.31 pg/ml; NGF, 300.89±33.51 pg/ml)(P<0.01). The inactive patients also had significantly increased serum levels of IL-13 and NGF as compared with normal controls (IL-13, 35.20±12.70 pg/ml; NGF, 111.40±32.54 pg/ml; P<0.05 and P<0.01, respectively). Spearman correlation analysis revealed that the serum IL-13 levels were correlated with disease activity index of SLE (SLEDAI), ESR and serum levels of C3 (r= 0.813, 0.504, -0.605, respectively). The serum NGF levels were also correlated with above markers (r=0.442, 0.338, -0.463, respectively). The serum levels of IL-13 and NGF had a positive correlation (r=0.506, P<0.01). It was suggested that IL-13 and NGF might be involved in the pathogenesis of SLE and closely correlated with disease activity.
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Objectives To investigate the serum levels of IL-13 and n erve growth factor (NGF) and their clinical significance in patients with system ic lupus erythematosus (SLE). Methods The serum levels of IL-13 and NGF were m easured by ELISA in 35 SLE patients and 15 normal controls. Results The serum levels of IL-13 and NGF in patients with active SLE were both significantly high er than those in patients with inactive SLE (IL-13: 92.69 ? 9.87 pg/mL versus 5 4.22 ? 9.31 pg/mL, P
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Objective To compare intraindividual higher-order aberrations(HOA)after implantation of aspherical and spherical intraocular lenses(IOL).Design Prospective,case-controlled study.Participants 33 patients(66 eyes)were between 50 and 76 years of ages.The patients with bilateral senile cataracts were operated with phacoemulsification and IOL implantation.All the patients had no other ocular pathologies and no previous ophthalmic surgery.Patients with keratometric astigmatism higher than-2.0 diopters(D)were excluded from the study.Methods Wavefront aberration measurements of 33 patients(66 eyes)after implantation of a spherical IOL(Canon Staar KS-3)in one eye and an aspherical IOL(Cannon Staar KS-3Ai)in the contralateral eye were analyzed.The eyes were randomly assigned to groups of the spherical IOL or the aspherical IOL.Third-order,4th-order,5th-order,total HOA root-mean-squares(RMS),and 4th-order spherical aberrations(Z40)were compared 3 months after surgery.Main Outcome Measures Visual acuity(VA),the best corrected visual acuity(BCVA),the centrality of IOL,refraction data,diameter of pupil size,3rd-order,4th-order,5th-order and total HOA RMS,and 4th-order spherical aberration(Z40)RMS were measured.Results Fourth-order aberration RMS(0.193?0.098?m),Z40 RMS(0.037?0.099?m),total HOA RMS(0.498?0.072?m)in the aspherical IOL group were lower than those with the spherical IOL(0.403?0.155?m,0.381?0.142?m,0.737?0.164?m,respectively)at 6mm pupil diameter.For 3rd-and 5th-order RMS at 6mm pupil diameter,total HOA RMS(at 5mm pupil diameter),no significant difference was found in both IOLs.Under mesopic conditions,ten patients(30%)with an aspheric IOL had better contrast sensitivity.The average myopic shift were(-0.29?0.09)D and(-0.87?0.16)D in the aspherical and spherical IOL groups at 6mm pupil diameter(P