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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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TongJi University School of Medicine has performed teaching reforms in the early stage of medical students through curriculum integration of life science,the introduction of PBL,and designing of comprehensive experiments to achieve comprehensive professional competence of medical students.The results showed that the training system is beneficial to the cultivation of students' clinical thinking and early medical professional competence.It has been recognized by both students and faculty.Preliminary practice has proved to be successful.
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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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Thought and consciousness of experimental design were carried in the experimental teaching and the classical physiological experiments were merged. Innovational design and practice were penetrated through the whole experimental class. After consulting literature, designing experi-ments, observing the phenomenon, dealing with the data and summurizing the results, the designers experienced the basic research procedure, got the innovate results, gained the chance to further study and improved the innovative capability.
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Experimental course for integrative life science curriculum is designed for medical students ,which integrates basic knowledge and skills of cellular biology,biochemistry,genetics,molec-ular biology,molecular genetics,and immunology disciplines. Tongji University School of Medicine start-ed to implement integration-based new curriculum system since 2010. This article discussed on teaching philosophy,teaching content,teaching methods,teaching team construction and explored the effect and significance of experimental course for integrative life science curriculum.
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According to content of ‘lipid metabolism’,we crafted a effective case for problem based learning (PBL) and practiced it in biochemistry course.In group discussion,students participated in discussion with great enthusiasm and initiative.During the discussion,students got a better understanding about lipid metabolism and clinical application of the knowledge.As teachers,we maily guided students during PBL,and at the same time,we also summed up the experience and explored how PBL be implemented for freshmen and sophomores who lack clinical practice.
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In medical physiology experiment courses, the students observe the functions and learn the rules of biological body, in order to improve their abilities of scientific observation, practice and reflection. Emotion control means willpower to manage emotion, which is the regular psychological response when we deal with issues. Integrating the emotion control education to professional education is imperative to elevate the general quality of medical students.
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Objective To observe the effects of subretinal transplantation of rat mesenchymal stem cells (rMSCs) on Sodium Iodate (SI)-induced retinal degeneration.Methods One hundred and twenty Brown-Norway (BN) rats were divided into three groups including SI injection group,rMSCs transplantation group and normal control group,each with 40 rats.The retinal degeneration was induced by caudal vein injection of SI.The retinal pigment epithelium(RPE)and neural retinal were evaluated by ocular fundus photograph,fluorescein fundus angiography (FFA),electroretinogram (ERG) and histological approach,and TUNEL(terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling).CM-Dilprelabeled primary rMSCs were transplanted into the subretinal space of Sl-induced rats.The survival,integration,and differentiation of rMSCs were observed between 14 day to 60 day after the transplantation.Results The rat retinal function was gradually reduced afterl4 days of SI injection,with a time-dependent manner.After the RPE cells were damaged,the outer segments of photoreceptors became disrupted and shortened until karyopyknosis.The nuclear morphology and positive TUNEL labeling indicated that the death of photoreceptor cells was apoptosis.After rMSCs transplantation,CM-DiI labeled donor cells were observed to be scattered in the subretinal space and expressed RPE cell markers.Average amplitude of bwave and Ops (oscillation potential) in ERG improved 27.80%,59.38% respectively after rMSCs transplantation.Conclusions Transplanted rMSCs can survive in subretinal space and differentiate into RPE cells,thus cure SI- induced retinal degeneration.