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manufacturing, industrial and agricultural. To improve the transduction efficiency of PMSCs, Lipofectamine 2000 was used to mediate adenoviral transduction. As shown in Fig. 2A, a very significant increase in the expression of EGFP in PMSCs was observed when the cells were transfected with Lipofectamine 2000, and a high-level viral titer resulted in enhanced expression of EGFP. Furthermore, flow cytometry was used to determine the transduction efficiency, in which 29.7% of PMSCs cells expressed EGFP at an MOI of 1500. This value was increased to 66.3% at an MOI of 3000 using Lipofectamine 2000 (Fig. 2B). These data were consistent with the results obtained from microscopic quantification. On the basis of these results, we selected an MOI of 3000 mixed with Lipofectamine 2000 at a ratio of 2.5×108 vp/µg for subsequent experiments. To improve the transduction efficiency of PMSCs, Lipofectamine 2000 was used to mediate adenoviral transduction. As shown in Fig. 2A, a very significant increase in the expression of EGFP in PMSCs was observed when the cells were transfected with Lipofectamine 2000, and a high-level viral titer resulted in enhanced expression of EGFP. Furthermore, flow cytometry was used to determine the transduction efficiency, in which 29.7% of PMSCs cells expressed EGFP at an MOI of 1500. This value was increased to 66.3% at an MOI of 3000 using Lipofectamine 2000 (Fig. 2B). These data were consistent with the results obtained from microscopic quantification. On the basis of these results, we selected an MOI of 3000 mixed with Lipofectamine 2000 at a ratio of 2.5×108 vp/µg for subsequent experiments.. All experimental procedures and protocols were approved by the Institutional Animal Care and Use Committee of Gyeongsang National University and performed in accordance with the Guide for the Care and Use of Laboratory Animals.. One of our most significant findings was the relationship between school type and the astigmatism prevalence. A challenging school refers to the one with skilled teachers, good infrastructure, much more investment from local government than non-challenging schools, and high university enrolment rates. Compared to a general school, a challenging school has a more competitive learning environment. This may encourage the enrollment of more talented students and may appeal to teachers with additional qualifications. Moreover, both teachers and students live stressful lives in the challenging school. There are more demands placed on teachers to produce students with good academic qualifications while there is more homework for students. Comparing astigmatism prevalence between academically challenging and regular schools, we discovered that the prevalence rates of myopic astigmatism and hyperopic astigmatism in academically challenging schools were significantly higher than those in regular schools. Furthermore, mild astigmatisms (≥ 0.50DC, P<0.001) between schools varied more significantly than moderate astigmatism (≥ 1.00DC, P=0.004). As mentioned above, students from academically challenging schools spend much more time reading and writing than those from regular school. We speculate that prolonged lowering of the upper eyelid during reading and writing can cause pressures on eyeballs, specifically corneas. These pressures may result in mild morphological changes on corneas which result in hyperopic and myopic astigmatism, especially mild astigmatism. Several research studies proved that eyelid's pressures such as granuloma, chalazia [30, 31], and congenital ptosis [32, 33] can contribute to astigmatism [34]. These findings support our inference. We therefore conclude that near-work activity may be one of the factors contributing to astigmatism. One of our most significant findings was the relationship between school type and the astigmatism prevalence. A challenging school refers to the one with skilled teachers, good infrastructure, much more investment from local government than non-challenging schools, and high university enrolment rates. Compared to a general school, a challenging school has a more competitive learning environment. This may encourage the enrollment of more talented students and may appeal to teachers with additional qualifications. Moreover, both teachers and students live stressful lives in the challenging school. There are more demands placed on teachers to produce students with good academic qualifications while there is more homework for students. Comparing astigmatism prevalence between academically challenging and regular schools, we discovered that the prevalence rates of myopic astigmatism and hyperopic astigmatism in academically challenging schools were significantly higher than those in regular schools. Furthermore, mild astigmatisms (≥ 0.50DC, P<0.001) between schools varied more significantly than moderate astigmatism (≥ 1.00DC, P=0.004). As mentioned above, students from academically challenging schools spend much more time reading and writing than those from regular school. We speculate that prolonged lowering of the upper eyelid during reading and writing can cause pressures on eyeballs, specifically corneas. These pressures may result in mild morphological changes on corneas which result in hyperopic and myopic astigmatism, especially mild astigmatism. Several research studies proved that eyelid's pressures such as granuloma, chalazia [30, 31], and congenital ptosis [32, 33] can contribute to astigmatism [34]. 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