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Objective To investigate the inhibiting effect of CGP77675 (CGP),a Src inhibitor,on epithelial-mesenchymal transition (EMT) of human retinal pigment epithelial (RPE) cells induced by transformation growth factor-β1 (TGF-β1).Methods Human RPE cell line (ARPE19 cells) was cultured in vitro and divided into control group,TGF-β1 group and TGF-β1 +CGP group.Corresponding agent was added into culture medium based on grouping.The morphology of the cells were examined under the optical microscope 3 days after culture.The expressions of EMT-related genes and proteins in the cells were detected by real-time quantitative PCR and Western blot,respectively,including fibronectin 1 (FN 1),and plasminogen activation inhibitor 1 (PAI1),and the expressions of zonula occludens protein 1 (ZO1) and cytoskeleton protein filamentous actin (F-actin) were detected by immunofluorescence staining.MTT assay was employed to evaluate the cell proliferation rate.The migration distance of the cells was measured by scratch test.Results The ARPE19 cells in the control group showed an epithelial-like morphology and F-actin and ZO-1 were expressed along cell membrane.In the TGF-β1 group,the cells appeared to be fibrous-like,and the fluorescence staining of F-actin was disordered and ZO-1 was discontinuous on the cell membrane.The cells in the TGF-β1 +CGP group remained to be an epithelial-like in shape with clear and complete expressions of F-actin and ZO-1.The relative expressions of FN1 mRNA and PAI1 mRNA in the cells were 0.211 ± 0.080 and 0.116±0.073,1.000±0.001 and 1.000±0.001,0.368±0.097 and 0.362±0.048 in the control group,TGF-β1 group and TGF-β1 +CGP groups,showing significant differences among the groups (F=33.14,82.92;both at P＜0.01),with the highest expressions ofFN1 mRNA and PAI1 mRNA in the TGF-β1 group (all at P＜0.05).The relative expressions of FN1 and PAI1 proteins were 0.166±0.055 and 0.327±0.066,1.000±0.001 and 1.000± 0.001,0.143 ± 0.030 and 0.260 ± 0.077 in the control group,TGF-β1 group and TGF-β1 + CGP group,with significant differences among three groups (F=181.90,48.85;both at P＜0.01),and the expressions FN1 and PAI1 proteins were significantly higher in the TGF-β1 than those in the control group and TGF-β1 +CGP group (all at P＜0.05).The cell proliferative rate in the TGF-β1+CGP group was (79.30±3.44) ％ and (54.80±7.39) ％ at the third day and seventh day after culture,which were significantly reduced in comparison with (99.50 ± 1.00)％ and (99.10±0.50)％ in the control group as well as (95.10±4.20)％ and (92.10±4.50)％ in the TGF-β1 group (all at P＜0.05).The migration distance was disappeared in the TGF-β1 group,and the scratch width was not obviously changed in the TGF-β1 +CGP group.Conclusions Src inhibitor can inhibit EMT process of ARPE19 cells induced by TGF-β1,indicating that Src signaling pathway may play a critical role in EMT of RPE cells.
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Department of Regenerative Medicine of Tongji University School of Medicine intro-duced the standardized question library construction-based separation of teaching from testing system into integrated life science course. By establishing the question library, professional teachers in the assessment center are responsible for making and correcting test papers of final exam for students. In addition to the separation of teaching and testing, regular quiz during the semester is also involved in the final grades of students. The results show that high-quality question library effectively promotes implementation of separation of teaching from testing. The question library construction is a dynamic and long-term task that requires real-time updates along with knowledge updates. This preliminary practice of separation of teaching from testing system in the integrated life sciences curriculum has proved to be useful for improving teaching style and the style of study significantly.
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To investigate the plasma thioredoxin-interacting protein ( TXNIP ) levels in different glucose tolerance groups and discuss the relationship between TXNIP and insulin resistance/β-cell dysfunction in diabetes and prediabetes, and to investigate the potential relationship between TXNIP and interleukin-1β( IL-1β) . According to oral glucose tolerance test, 93 participants were divided into 3 groups:diabetes mellitus group, prediabetes group, and normal glucose tolerance group. Plasma TXNIP, IL-1β, and other biochemical indices were measured. The relationship between TXNIP and glucose, IL-1β, homeostasis model assessment for insulin resistance ( HOMA-IR) , and homeostasis model assessment forβcell function ( HOMA-β) were analyzed by using multiple linear regression techniques and Pearson’s linear correlation analysis. Plasma TXNIP level was higher in prediabetes group compared with normal glucose tolerance group, but lower in prediabetes group compared with diabetes mellitus group[(355. 35±31. 88 vs 274. 36±33. 86, 426. 16±63. 15)pg/ml, P<0. 01 or P<0. 05]. TXNIP was positively correlated with IL-1βand HOMA-IR, but negatively correlated with HOMA-β. Multiple linear regression analysis indicated that IL-1βexerted significant influence on TXNIP ( P<0. 05 ). Plasma TXNIP level is affected by blood glucose concentration. There is a close relationship between TXNIP and IL-1β. In prediabetes patient, the TXNIP levels have already been raised.
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Objective The different distribution and clinical significance of mean platelet volume (MPV) in the healthy normoglycemic and impaired fasting glucose (IFG) individuals were discussed.Methods The 499 individuals including 184 male and 315 female,who had undergone health checks in Tianjin Huanhu Hospital during May and July 2012 were studied retrospectively.Average age is forty-five ( thirty-five to eighty).Subjects were categorized into four groups according to fasting plasma glucose ( FPG) levels:G1 (3.89 mmol/L≤FPG<5 mmol/L, n=125),G2(5 mmol/L≤FPG <5.5 mmol/L, n=121), G3(5.5 mmol/L≤FPG<6.1 mmol/L, n=142), and G4(6.1 mmol/L≤FPG<7 mmol/L, n=111).G1, G2, and G3 are defined as normal FPG groups and G 4 is defined as IFG group.Eighty-nine cases in the same age patients with type II diabetes mellitus group ( G5 ) were observed at the same time.Results The MPV increased with the increasing FPG levels in the following order:G1(8.62 ±0.77) fl, G2 (8.85 ±0.80) fl, G3(8.90 ±0.69) fl,G4(9.14 ±0.78) fl and G5(12.03 ±1.42) fl.MPV[(12.03 ±1.42) fl] of type Ⅱdiabetes mellitus group(G5) was higher than that in the IFG group (G4)[(9.14 ±0.78) fl] and normal FPG groups[G1(8.62 ±0.77) fl,G2(8.85 ±0.80) fl,G3(8.90 ±0.69) fl] (F=12.773,P<0.01);MPV of the IFG group [ ( 9.14 ±0.78 ) fl ] was significantly higher than that in normal FPG groups [ G1 (8.62 ±0.77) fl,G2(8.85 ±0.80) fl,G3(8.90 ±0.69) fl] (F=12.773,P<0.01 for G4 vs.G1 and G2, P<0.05 for G4 vs.G3) ;MPV in the high-normal glucose group (G3) [(8.90 ±0.69) fl] was obviously higher than that in the low-normal glucose group (G1) [(8.62 ±0.77) fl] (F=12.773,P<0.05);MPV was positively associated with FPG in normal FPG groups ,IFG group and type Ⅱ diabetes mellitus group (G1-3:r=0.22, P<0.05;G4:r=0.26, P<0.01;G5:r=0.29, P<0.01).Conclusions Significant difference of MPV was observed in population of different FPG levels.Especially, MPV in IFG group was evidently higher than that in normal FPG group and was positively associated with FPG levels.