1.Expressions of interleukin-4,5 and 13 in ocular surface with different types of allergic conjunctivitis
Juan, YAO ; Bing, LI ; Xiaofen, ZHENG
Chinese Journal of Experimental Ophthalmology 2015;33(10):924-929
Background The pathogenesis of allergic conjunctivitis has not been clearly established.Current researchers indicate that interleukin-4 (IL-4), IL-5 and IL-13 may play an important role in allergic conjunctivitis.But whether the roles of these inflammatory factors are same in different types of allergic conjunctivitis remains unclear.Objective This study was to investigate the expressions of IL-4, IL-5 and IL-13 in ocular surface with different types of allergic conjunctivitis.Methods A prospective cohort study was designed.Eighty individuals were recruited in Shanxi Eye Hospital from April 2013 to September 2014, including 20 patients with vernal keratoconjunctivitis (VKC), 20 patients with seasonal allergic conjunctivitis (SAC), 20 patients with perennial allergic conjunctivitis (PAC) and 20 normal healthy subjects.Surficial tissues were binocularly scraped using disinfected scraper from upper eyelid conjunctiva, and 4 μl of tear fluid was obtained with capillary tube.The expressions of IL-4, IL-5 and IL-13 protein and mRNA in the conjunctival epithelial cells were detected by immunohistochemistry and real-time fluorescence quantitative PCR.The IL-4, IL-5 and IL-13 concentrations in tear fluid were assayed by Luminex method.This study complied with Declaration of Helsinki and the research protocol was approved by the Shanxi Eye Hospital Ethics Committee.Written informed consent was obtained from each subject prior to entering the cohort.Results IL-4, IL-5 and IL-13 were positively expressed in cytoplasm of conjunctival epithelial cells in the VKC group,SAC group and PAC group,but the expressions of IL-4,IL-5 and IL-13 were absent in the normal control group.The relative expression levels of IL-4 mRNA were 4.11±1.24,2.71±0.71 and 2.00±0.80;the relative expression levels of IL-5 mRNA were 4.02±0.43,2.07±0.45 and 1.47±0.50;and the relative expression levels of IL-13 mRNA expression levels were 6.44±0.66,4.35±1.26 and 2.39±0.86 in the VKC group,SAC group and PAC group, showing significant differences among the 4 groups (F =51.32,220.18,162.49, all at P<0.01).The relative expression levels of IL-4,IL-5 and IL-13 mRNA were significantly higher in the VKC group than those in the SAC group and PAC group;and those in the SAC group were significantly elevated in comparison with the PAC group (all at P<0.05).No IL-4, IL-5 and IL-13 were detected in the tear fluid in the normal control group;while the concentrations of IL-4,IL-5 and IL-13 in the tear fluid were (14.06±3.50), (10.88±1.82) and (34.28±8.42) pg/ml in the VKC group,and (7.71 ±0.65), (5.10± 1.33), (23.77±6.29) pg/ml in the SAC group as well as (3.30± 1.50) pg/ml, (2.43± 1.28) pg/ml and (17.67 ± 4.28) pg/ml in the PAC group, showing significant differences among the 3 groups (F =200.29,260.49,128.23, all at P<0.01).IL-4, IL-5 and IL-13 concentrations in the tear fluid were significantly higher in the VKC group than those in the SAC group and PAC group,and those in the SAC group were significantly raised in comparison with the PAC group (all at P<0.01).Conclusions IL-4,IL-5 and IL-13 participate in the pathogenesis of multiple allergic conjunctivitis,but their expressions in the ocular surficial tissue are discriminatory in different types of allergic conjunctivitis.
3.cDNA Cloning, Expression Analysis of Slc24a5 and Its Relationship With Melanin Deposition in Chicken
Juan LUO ; Wei ZHANG ; Bing YU ; Xiaoxiang HU ; Ning LI
Progress in Biochemistry and Biophysics 2008;35(1):69-76
Melanin is a kind of biopolymer, composed of a complex quinine/indole-quinone derived mixture which is produced in melanocytes from tyrosine. More than 100 genes related to melanin deposition have been found in mouse. Slc24a5 (solute carrier family 24, member 5) is a novel gene first cloned in zebrafish. And it has been confirmed that the Slc24a5 is involved in controlling the melanin deposition in zebrafish. Here the full length of the chicken slc24a5 mRNA sequence, its expression profile and a discussion of its relationship to melanin deposition in chicken were reported. Chicken Slc254a5 has a CDS of 1 269bp, predicting a protein of 423 amino acids which is about 80 amino acids shorter than in human and mouse. It has 9 exons and 8 introns and spans more than 1lkb of genome sequence. The quantitative real-time PCR confirmed that the chicken Slc24a5 was highly expressed in the eye of White Leghorn and Beijing Fatty Chickens, and in the eye, skin and muscle of Silky. The relative expression in Silky eye is more than two times that of White Leghorn. The relative expression in Silky skin is about 70 times that of White Leghorn and about 15 times that of Beijing Fatty Chickens. The relative expression in Silky muscle is about 15 times that of White Leghorn and about 3 times that of Beijing Fatty Chickens. And the expression result is in accordance with the melanocyte and melanin deposition in these tissues.
5.The influence of adenovirus latent infection on glutathione and antioxidative enzymes in rat alveolar epithelial cells
Yi FANG ; Bing LI ; Juan CHEN ; Qicai LIU ; Pixin RAN
Chinese Journal of Microbiology and Immunology 2008;28(5):416-420
Objective To analyze the influence of adenovirus latent infection on gamma-glutamylcysteine systhetase(γ-GCS) in rat alveolar epithelial cells. Methods The rat alveolar epithelial cells were stably transfected with the plasmid pE1Aneo and control plasmid. Glutathione(GSH) contents, the activity of γ-GCS were detected in oxidant stress. Then the leuel of protein expression, mRNA expression, and promoter transcriptional activity of glutamate-cysteine ligase catalytic subunit(GCLC) were further detected. Results GSH contents decreased because of adenovirus E1A expression in oxidant stress. E1A repressed the expression and activity of γ-GCS, messenger RNA expression, and promoter transcriptional activity of GCLC. Conclusion Adenovirus E1A decreased the activity of γ-GCS probably by repressed promoter transcriptional activity of GCLC. As a result, GSH contents were downregulated in oxidant stress. Thus Adenovirus latent infection amplified the oxidant/antioxidant imbalance in rat alveolar epithelial cells in oxidants stress, which may be an important mechanism of COPD.
6.miR-496 over-expression inhibits growth and metastasis in colon cancer cells
Jingyi ZHOU ; Bing ZHONG ; Li LIU ; Juan WANG
Chinese Journal of Pathophysiology 2016;32(10):1815-1823
[ ABSTRACT] AIM: To investigate the effect of miR-496 over-expression on the growth and metastasis of colon cancer cells and its molecular mechanism .METHODS:The proteins interacting with miR-496 were screened by bioinfor-matic method.The levels of miR-496, CTNNB1 mRNA and β-catenin protein in colon cancer cell lines , HT29, HCT116 and SW480, and normal colonic epithelial cell line NCM 460 were detected by real-time PCR and Western blot .HT29, HCT116 and SW480 cells were transfected with miR-496 mimics using Lipofectamine 2000 and named as HT29-miR-496 mimics, HCT116-miR-496 mimics and SW480-miR-496 mimics cells, respectively, and the cells transfected with the scramble served as negative control .The cell viability, lactate dehydrogenase (LDH) leakage, and colony formation and metastatic abilities were determined by MTT assay , LDH assay, colony formation assay and Transwell method , respective-ly.The promoter activity of miR-496 was measured using luciferase reporter gene assay .The protein levels of β-catenin, eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1), p-4E-BP1, low-density lipoprotein receptor-related protein 6(LRP6), p-LRP6, MMP-7, MMP-9, MMP-13 and TIMP-2 were monitored by Western blot.RESULTS:Endog-enous miR-406 interacted with β-catenin was found in the colon cancer cells .Low miR-496 expression in the HT29, HCT116 and SW480 cells and high miR-496 expression in NCM460 cells were detected.In contrast, high β-catenin ex-pression was found in the HT29, HCT116 and SW480 cells and low β-catenin expression was observed in the NCM460 cells.Compared with control group , the cell viability, colony formation rate and the number of metastatic cells remarkably decreased in the HT29-miR-496 mimics, HCT116-miR-496 mimics and SW480-miR-496 mimic cells (P<0.05).The promoter activity of miR-496 was significantly increased in colon cancer cells transfected with miR-496 mimics, and was 1.75, 2.04 and 1.61 times as high as control group .miR-496 over-expression inhibited β-catenin levels, and p-4E-BP1 and p-LRP6 protein levels were also reduced .siRNA-or over-expressed miR-496-mediated β-catenin down-regulation in-hibited MMP-7 and MMP-9 expression , but promoted TIMP-2 expression .CONCLUSION:The expression level of miR-496 in the colon cancer cells is low , but in the normal colonic epithelial cells is high .miR-496 over-expression inhibits the protein levels of MMP-7 and MMP-9, and promotes the protein expression of TIMP-2 via inhibiting Wnt/β-catenin path-way, thus suppressing malignant phenotype in the colon cancer cells .
7.Adenovirus latent infection enhances the oxidant/antioxidant imbalance in rat alveolar epithelial cells
Yi FANG ; Bing LI ; Juan CHEN ; Qicai LIU ; Pixin RAN
Chinese Journal of Pathophysiology 1989;0(06):-
AIM:To observe the influence of adenovirus latent infection on the oxidant/antioxidant balance in rat alveolar epithelial cells.METHODS:The rat alveolar epithelial cells were stably transfected with the plasmid pE1Aneo and control plasmid pneo.GSH and MDA contents,the activities of major anti-oxidative enzymes including SOD,CAT,GPx,GST and ?-GCS were detected in oxidant stress.RESULTS:Adenovirus E1A expression repressed the activity of ?-GCS,and decreased GSH contents in oxidant stress.As a result,the activity of GPx and GST was decreased.The contents of MDA maintained high in oxidant stress.CONCLUSION:Adenovirus latent infection amplifies the oxidant/antioxidant imbalance in rat alveolar epithelial cells in oxidants stress,and adenovirus E1A protein decreases the activity of ?-GCS,which plays an important role in this process.
8.Attenuated store-operated Ca2+ entry in mesenteric artery endothelial cells of aged rat
Xia WANG ; Jie LI ; Yan ZHU ; Juan DU ; Bing SHEN
Chinese Journal of Geriatrics 2015;34(4):434-437
Objective To observe attenuated store-operated Ca2+ entry in superior mesenteric artery endothelial cells of aged rat.Methods The pressure myograph was applied to measure the vasodilation of superior mesenteric artery induced by bradykinin (BK) in aged versus non-elder rats.Primarily cultured MAECs were treated by thapsigargin (TG) and BK to deplete intracellular Ca2+ stores for comparing SOCE.SOCE of MAECs was detected by calcium imaging technology and was compared between aged and non-elder rats.The expression levels of Orai 1 and stromal interaction factor 1 (STIM 1) were observed by immunofluorescence method.Results Compared with the nonelder rats (70.0 ± 4.1 %),BK-induced vasodilation in aged rats (27.0 ± 4.9)% was declined by (60.7±4.3) %.SOCE induced by BK and TG in primarily cultured MAECs was attenuated by 37.6% and 39.2% in aged rats (1.71±0.18 and 1.06±0.03) respectively as compared with non-elder rats (2.72±0.39 and 1.75±0.06).The expressions of SOCE's components Orai 1 and STIM 1 in MAECs were decreased obviously in aged rats than in non elder rats.Conclusions SOCE of MAECs and SOCE-induced vasodilation are significantly attenuated in aged rats.The decreased expression level of Orai 1 and STIM 1 may contribute to this alteration.
9.Relationship between Clinical Neuro-Electrophysiology and Prognosis in Children with Guillain-Barr? Syndrome
chang, PENG ; xiao-mei, SHU ; bing-zhu, YANG ; juan, LI
Journal of Applied Clinical Pediatrics 1992;0(05):-
ObjectiveTo explore the relationship between the changes of neuro-electrophysiology and prognosis in children with Guillain-Barr? syndrome(GBS).MethodsThirty-eight children with GBS were divided into group A(rapid recovery,n=16) and group B(slow recovery,n=22) according to the time required for podosoma motor function recovery,at the same time,they were divided into the better prognosis group(n=22) and the worse prognosis group(n=16),for analyzing the difference between group A and B in terms of age,preceding infections,maximal Hughes grades and neuro-electrophysiology including motor conduction velocity(MCV),distal complex muscle action potential(dCMAP) and F wave,and investigating the related factors with the prognosis of GBS.Results1.MCV of tibial nerve was(40.2?2.53) m/s and(33.4?2.46) m/s in group A and group B,respectively;MCV of peroneal nerve was(45.2?3.23) m/s and(38.3?2.16) m/s in group A and group B,respectively,and the difference between group A and group B was significant(Pa0.05);abnormal rate of F wave(68.42%) was higher than abnormal rate of MCV(42.11%) and dCMAP(42.11%)(Pa
10.Inhibition of physiological concentration of glucocorticoids on LPS-induced inflammation in rat alveolar epithelial cells
Juan CHEN ; Jiandong LUO ; Bing LI ; Dongting ZOU ; Pixin RAN
Chinese Pharmacological Bulletin 2003;0(08):-
Aim To investigate the role of physiological concentration of glucocorticoids on the inflammation mediator IL-6 expression in response to LPS in rat alveolar epithelial cells(CCL149).Methods The CCL149 were treated with LPS,H2O2 and glucocorticoid respectively.Flammtory mediator IL-6 protein expression was measured with ELISA,and the activity of histone deacetylase(HDAC) was measured using colorimetric HDAC activity assay kit.Results IL-6 protein levels were increased in cells exposed to 10 mg?L-1 LPS.Hydrocortisone decreased IL-6 protein expression induced by LPS.Such effect of hydrocortisone was blunt by HDAC inhibitor trichostatinA treatment(10 ?g?L-1).LPS decreased HDAC activity.Hydrocortisone increased HDAC activity.The expression of IL-6 protein induced by LPS was further enhanced by H2O2 treatment.Pretreatment with H2O2 resulted in the inhibition of antiflammtion effect of glucocorticoids.Conclusion Physiological concentration of glucocorticoids could suppress inflammatory response,and this effects requires recruitment of HDAC.Oxidants such as H2O2 may cause the failure of glucocorticoids to function effectively,and the reason may be related to the reduction of HDAC activity.This mechanism may contribute to the pathogenesis of pulmonary disorder.