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Objective:To observe the changes of follistatin-like protein 1 (FSTL1) in serum of patients with proliferative diabetic retinopathy (PDR).Methods:Twenty PDR patients confirmed by clinical examination and 20 normal people were included in the study. Human retinal vascular endothelial cells (HRCEC) were divided into HRCEC blank control group, 3 h hypoxia group, 6 h hypoxia group. Human umbilical vein endothelial cell (HUVEC) were divided into HUVEC blank control group, 3h hypoxia group, 6h hypoxia group. Real-time quantitative PCR (RT-PCR) and ELISA were used to determine the expression of FSTL1, TGF-β, VEGF, connective tissue growth factor (CTGF) mRNA and protein in peripheral blood and cells of all groups from all subjects.Results:The expressions of FSTL1, TGF-β1, CTGF, VEGF mRNA in blood samples of patients with PDR were 1.79±0.58, 0.97±0.21, 1.85±0.69 and 1.38±0.44. The expressions of FSTL1, TGF-β1 protein were 1.19±0.50, 0.71±0.24 ng/ml and 734.03±116.45, 649.36±44.23 ng/L. Compared with normal people, the differences were statistically significant ( tmRNA=0.90, 0.21, 2.85, 1.77; P=0.00, 0.00, 0.04, 0.02. tprotein=1.88, 7.68; P=0.00, 0.02). The cell viability of HRCEC cells in the 3 h hypoxia group and the 6 h hypoxia group were 0.66±0.05 and 0.64±0.04, respectively. Compared with the blank control group, the difference was statistically significant ( F=13.02, P=0.00). The cell viability of HUVEC cells in the 3 h hypoxia group and the 6 h hypoxia group were 0.63±0.06 and 0.68±0.06, respectively. Compared with the blank control group, the difference was statistically significant ( F=26.52, P=0.00). Comparison of FSTL1, TGF-β1, CTGF, and VEGF mRNA expression in HRCEC blank control group and 3 h hypoxia group, the differences were statistically significant ( F=14.75, 44.93, 85.54, 6.23; P=0.01, 0.00, 0.00, 0.03). Compared with the HRCEC blank control and 3 h hypoxia group, the expressions of FSTL1 and TGF-β1 protein were statistically significant ( P<0.05). There was a statistically significant difference in TGF-β1 protein expression in the hypoxic 6 h group ( P=0.03) and no significant difference in FSTL1 protein expression ( P=0.68). Comparison of FSTL1, TGF-β1, CTGF, and VEGF mRNA expression in HUVEC blank control group and 3h hypoxia group, the differences were statistically significant ( F=19.08, 25.12, 22.89, 13.07; P=0.00, 0.00, 0.00, 0.01). Immunofluorescence staining results showed that FSTL1, TGF-β1, CTGF, and VEGF proteins were positively expressed in cells in the 3h hypoxia and 6h hypoxia groups. Conclusion:The expression of FSTL1 gene and protein in serum of PDR patients was significantly higher than that of normal people.
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Objective To construct the connective tissue growth factor (CTGF) recombinant interference vector (shRNA) and observe its inhibitory effect on the expression of endogenous CTGF in retinal vascular endothelial cells.Methods The human CTGF shRNA was constructed and the high-titer CTGF shRNA lentivirus particles was acquired via three-plasmid lentivirus packaging system to infect retinal vascular endothelial cells.The optimal multiplicity and onset time of lentivirus infection were identified by tracing down the red florescent protein in interference vector.The cells were classified into three groups:blank control group,infection control group and CTGF knockdown group.The differences in cells migrating ability was observed through Transwell allay.The mRNA and protein expression of CTGF,fibronectin,a-smooth muscle actin (α-SMA) and collagen Ⅰ (Col Ⅰ) were quantified through real-time PCR testing and Western blot system.Data between the three groups were examined via one-way analysis of variance.Results The result showed that an optimal multiplicity of 20 and onset time of 72 hours were the requirements to optimize lentivirus infection.Transwell allay result showed a contrast in the number of migrated cells in the CTGF knockdown group and that in the blank control group and infection control group (F=20.64,P=0.002).Real-time PCR testing showed a contrast in related gene expression (CTGF,fibronectin,α-SMA and Col Ⅰ) in the CTGF knocked-down group and that in the blank control group and infection control group (F=128.83,124.44,144.76,1 374.44;P=0.000,0.000,0.000,0.000).Western blot system showed the statistical significance of the contrasted number of related protein expression (CTGF,fibronectin,α-SMA and Col Ⅰ) in the knockdown group and that in the blank control group (F=22.55,41.60,25.73,161.68;P=0.002,0.000,0.001,0.000).Conclusion The success in producing CTGF shRNA lentivirus particle suggests that CTGF shRNA lentivirus can effectively knock down CTGF expression.
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Intravitreal injections of anti-vascular endothelial growth factor (anti-VEGF) drugs,including monoclonal antibodies (such as bevacizumab and ranibizumab) and fusion protein agents (such as aflibercept and conbercept) have been proven to be effective in the treatment of wet age-related macular degeneration (wAMD).However,there are still some patients with poor efficacy,such as no response to initial treatment or poor response,and even relapse during the course of treatment.In view of the different targets and molecular characteristics of anti-VEGF drugs,the switch of anti-VEGF drugs and the adjustment of delivery pattern,dosages and intervals have been the strategies to cope with the poor efficacy in clinic.However,there are some differences in the results of current studies.Overall,the recovery of retinal anatomical outcome achieves more benefits,and it is relatively difficult to improve visual acuity.To determine which regimen would get the biggest benefits,a large number of randomized controlled clinical trials and long study period will be needed.
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Objective To observe RNA-Seq analysis of gene expression profiling in human retinal vascular endothelial cells after anti-vascular endothecial growth factor (VEGF) treatment.Methods Cultured the retinal vascular endothelial cells in vitro and logarithmic growth phase cells were used for experiments.The cells were divided into VEGF group and VEGF combined with anti-VEGF drugs group.The VEGF group cells were treated with 50 ng/ml VEGF for 72 h to simulate the high VEGF survival conditions of vascular endothelial cells in diabetic retinopathy.VEGF combined with anti-VEGF drug group cells was treated with 50 ng/ml VEGF and 2.5 μg/ml anti-VEGF drugs for 72 h to imitate the microenvironment of cells following the anti-VEGF drugs treatment,and whole transcriptome sequencing approach was applied to the above two groups of cells through RNA-Seq.Now with biological big data obtained as a basis,to analyze the differentially expressed genes (DEGs).And through enrichment analysis to explain the differential functions of DEGs and their signal pathways.Results The gene expression profiles of the two groups of cells were obtained.Through analysis,328 DEGs were found,including 194 upregulated and 133 downregulated ones.The functions of DEGs were influenced by regulations over molecular biological process,cellular energy metabolism and protein synthesis,etc.Among these genes,SI,PRX and HPGD were related to protein synthesis,BIRCT to cellular apoptosis,and ABLIM 1 and CRB2 to retinal development,and ABCG 1,ABCA9 and ABCA12 were associated with the cholesterol of macrophage and the transfer of phospholipid.GO enrichment analysis showed that DEGs mainly act in three ways:regulating biological behavior,organizing cellular component and performing molecular function.Pathway enrichment analysis showed that gene expressions of the two cell groups were differentiated in ECM receptor pathway,and Notch,mitogen-activated protein kinase,transforming growth factor (TGF)-β and Wnt signal pathways.Among them,the gene expression in TGF-β signal pathway attracts most attention,where the DEGs,such as CAMK2B,COL3A1,CYGB,PTGER2 and HS6ST2,among others,were closely related to fibrosis process.Conclusion The anti-VEGF drugs may enhance the expression ofCAMK2B,COL3A1,CYGB,PTGER2 and others genes related to TGF-β signal pathway and aggravate retinal fibrosis disease.
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The establishment of a scientific salary system is a key measure to improve the incentive and re-straint mechanisms, adjust and optimize the relations of the production, release the reform “bonus”, and is also an important health care reform today. In order to establish a scientific salary system of medical personnel adapting to the health industry characteristics, the paper makes an investigation on the salary level in the years of 2010—2012 in Shanghai, and analyzes the main problems of the present performance pay policy. On this basis, starting from the point of the health industry characteristics, the paper builds the salary system of medical personnel containing the comprehensive budget management, theoretical workload for approval, authorized personnel, performance appraisal and distribution, structural proportion within the industry, fund distribution management through the establishment of the salary level’s average wage and social linkage mechanism, and builds a theoretical framework on salary system of medical personnel in Shanghai.
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Objective To conduct a health check-up intention analysis to understand residents' altitude to health eheck-up. Methods A total of 3600 residents living in Shanghai completed the selfdesigned questionnaire. Results About 35.2% participants received health check-up onee per year. In 36 to 60 or > 60 age group, the figure was increased up to 62.6% or 50. 4%, respectively. Public institution employees and those with medical insurance were found to have the highest rate of health checkup. Conclusion Our investigation results suggest that government leadership,health management model and contents, and personnel training should be improve.
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Objective To explore the status of Cryptosporidium infection among injection drug abusers(IDUs) based on the purified recombinant CP23 protein antigen.Methods The serologic antibody response of Cryptosporidium infection was examined by indirect ELISA in the collected 588 serum episodes of IDUs and 384 cases of healthy persons. Results Among 588 cases of IDUs, 69.90% of them were serologic antibody-positive, compared with 29.43% of the healthy persons, with a significant difference(P